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SCRIPPS FLORIDA ANNUAL REPORT FOR THE YEAR ENDING JUNE 30, 2015

PART 1: GRANT AWARDS AND SCIENTIFIC ACHIEVEMENTS

A. NEW GRANTS Scripps Research Institute Scientists Awarded $7.9 Million to Develop Artificial Immune System Scientists from both campuses of The Scripps Research Institute (TSRI) have been awarded a total of $7.9 million from the Defense Advanced Research Projects Agency (DARPA) of the U.S. Department of Defense. The two teams will build what is, in essence, an artificial immune system, comprising vast “libraries” of different types of molecules from which will emerge individual compounds to detect or neutralize an array of biological and chemical threats. Under the auspices of DARPA’s new Fold F(x) Program, the Jupiter, Florida team, led by Professor Tom Kodadek and Assistant Professor Brian Paegel, will receive $5.7 million; the La Jolla, California team, led by Professor Floyd Romesberg, will receive $2.2 million. Developing New Libraries In Jupiter, Kodadek, Paegel and their colleagues will develop libraries of functional compounds and engineer highly automated strategies for rapid synthesis, screening and production. These libraries will contain molecules each tagged with a DNA “barcode” that uniquely identifies the molecules’ chemical structure. “We hope to create chemical libraries and screening platforms that are truly revolutionary in their capabilities,” Kodadek said. For Paegel, the DARPA grant will expand his lab’s current program in drug discovery technology development. His team has developed a microfluidic circuit that screens single compounds suspended on artificial beads, processing more than 200,000 compounds in a matter of hours. “We envision next-generation small molecule discovery as a distributed enterprise, not just limited to facilities like our molecular screening center in Jupiter,” Paegel said. “Our ultra-miniaturized approach will make this vision a reality.” Evolving New Functions In La Jolla, Romesberg and his colleagues will develop variants of oligonucleotides—short, single-stranded DNA or RNA molecules—modified to be both stable and to have increased functionality. The team will leverage a system known as SELEX (Systematic Evolution of Ligands by Exponential Enrichment) to evolve novel function molecules. “We plan to modify the classical SELEX methodology with two innovations from our previous work,” Romesberg said. The first innovation, developed by Tingjian Chen, a postdoctoral fellow in the Romesberg lab, is a DNA polymerase evolved to recognize nucleotides with modified sugars, which impart the

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corresponding oligonucleotide polymers with increased thermal stability and resistance to enzymes that typically degrade oligonucleotides. The second innovation is an unnatural base pair, developed as part of the team’s recent expansion of the genetic alphabet, which can be modified with linkers to site-specifically attach different functionality to oligonucleotides. The combined technologies should allow for the evolution of novel biopolymers that are both stable and possess virtually any desired binding or catalytic activity. Scripps Florida Scientists Awarded $3.5 Million to Expand Development of New Diabetes Therapies Scientists from the Florida campus of The Scripps Research Institute (TSRI) have been awarded $3.5 million from the National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health to accelerate development of a new class of anti-diabetic compounds. Patrick R. Griffin, chair of the Department of Molecular Therapeutics at Scripps Florida and a leader in the field, is the principal investigator of the new five-year grant. “Effective management of diabetes and the complications associated with the disease remains a significant medical challenge,” Griffin said. “Due to significant safety concerns, a class of drugs that have proven effective at improving the body’s response to insulin (insulin sensitizers known as glitazones) has essentially been removed from the arsenal of therapeutics used to treat type 2 diabetes.” Over the past decade, the Griffin lab along with the Kamenecka lab has focused on the molecular details of the mode of action of insulin sensitizers. Using this information, the scientists have made significant advances in developing drug candidates targeting a receptor known as peroxisome proliferator-activated receptors gamma (PPARG). These drug candidates inhibit the receptor, a unique mode of action compared to the glitazones. Diabetes affects more than 29 million people in the United States, according to the American Diabetes Association 2012 report. Between 2010 and 2012, the incidence rate was about 1.7-1.9 million per year, and in 2013, the estimated direct medical costs were $176 billion. This new award will fund deep dissection of the molecular mechanism of the new class of compounds developed at TSRI, and this information will help pave the path toward clinical development. In addition, the Griffin lab, in collaboration with researchers at the University of Toledo, will look at the effects of these compounds on bone, an emerging safety issue with the glitazones. The number of the new grant is 1R01DK105825.

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Scripps Florida Scientists Win $3.3 Million Grant to Accelerate Development of Treatments for Intellectual Disability, Autism, Epilepsy Scientists from the Florida campus of The Scripps Research Institute (TSRI) have been awarded $3.3 million by the National Institutes of Health (NIH) to identify biomarkers to accelerate drug development for disorders including autism spectrum disorder, epilepsy and some types of intellectual disability. Gavin Rumbaugh, a TSRI associate professor, is the principal investigator of the new five-year project. “Our long-term goal is to increase the success rate of therapies translated from animal models to patients,” Rumbaugh said. “By validating biomarkers in mice and using this information in combination with pharmacological or genetic treatment strategies, we hope to create a set of tools and methods that can be used successfully to develop new therapeutics.” Rumbaugh has been a pioneer in the study of Syngap1, one of the most commonly disrupted genes in patients with sporadic developmental disorders of the brain. His work in animal models has shown that life-long cognitive disruptions are caused by isolated damage to developing neurons in the forebrain (in humans, the forebrain is responsible for higher cognitive processes, such as language and reasoning). Rumbaugh and his colleagues plan to validate several highly quantifiable biomarkers of brain damage that occur in these animal models during a critical period of early development. Because abnormal cognition in these models can be traced to this early developmental window, these measures have the potential to provide a roadmap of cognitive ability to guide drug design. The number of the grant, from the NIH’s National Institute of Mental Health, is 1R01MH108408. Scripps Florida Scientists Win $2.4 Million to Expand Development of New Pain Therapies Scientists from the Florida campus of The Scripps Research Institute (TSRI) have been awarded $2.4 million from the National Institute on Drug Abuse of The National Institutes of Health to expand development of new pain medications with fewer side effects than those currently available. TSRI Professor Laura Bohn, who has been a leader in the development of pain therapies, will be the principal investigator of the new five-year grant. “We are developing substitutes for narcotic pain killers with less risk for overdose and fewer side effects,” Bohn said. “The new grant enables us to study how these potential drugs, which utilize the same biological target as morphine, fundamentally differ from the current pain medications in how they engage neuronal signaling.” Adverse side effects of current opioid drugs such as morphine and oxycodone can be serious and include respiratory suppression, constipation and addiction. According to the U.S. Centers for

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Disease Control, nearly two million Americans abused prescription painkillers in 2013; almost 7,000 people are treated each day in hospital emergency rooms for abuse of these drugs. While the new compounds under development activate the same receptor as morphine—the mu opioid receptor or MOR—they do so in a way that avoids recruiting the protein beta-arrestin 2. Genetic studies have shown that animal models lacking beta-arrestin 2 experience robust pain relief with diminished side effects. “The difference in the way that these new compounds work results in greater pain relief without as much respiratory suppression (overdose risk) and persistent constipation in preclinical studies,” said Bohn. “We are hoping to dial out dependence liabilities as we pursue bringing these drugs to clinical trials.” The number of the new grant is 1R01DA038964. Scripps Florida Scientists Win Grant to Uncover Ways to Erase Toxic PTSD Memories Scientists from the Florida campus of The Scripps Research Institute (TSRI) have been awarded $2.3 million from the Department of Health and Human Services of the National Institutes of Health to better understand how memories are stored in the hopes of eventually being able to treat posttraumatic stress disorder (PTSD) by erasing traumatic memories without altering other, more benign ones. Courtney Miller, a TSRI associate professor, is the principal investigator for the new five-year study. “We hope this new study will make a significant contribution to the goal of developing new and more effective treatments for mental illness,” Miller said. While literally thousands of mechanisms for how a memory initially forms have been identified, only a few mechanisms are known for how the brain stores these memories for weeks to years. To produce a memory, a lot has to be done, including the alteration of the structure of nerve cells via changes in the dendritic spines—small bulb-like structures that receive electrochemical signals from other neurons. Normally, these structural changes occur via actin, the protein that makes up the infrastructure of all cells. Miller is investigating the possibility that microRNAs, naturally occurring small RNAs that act to suppress the production of proteins, may be capable of coordinating the complexity required for the brain to maintain this actin-based structural integrity of a long-lasting memory. “Our study will investigate the microRNA profile of a PTSD-like memory, with the idea that the persistence of a traumatic memory is maintained by the recruitment of a unique set of microRNAs within the amygdala—the brain’s emotional memory center and a critical participant in PTSD,” Miller said.

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An understanding of how the brain actually stores these toxic memories should result in the development of new targets that can then be exploited to selectively target harmful memories, as in the case of PTSD, or to preserve fading memory, such as with age-related cognitive decline. In 2013, Miller and her colleagues were able to erase dangerous memories associated with drugs of abuse in mice and rats, without affecting other more benign memories. That surprising discovery, published in the journal Biological Psychiatry pointed to a clear and workable method to disrupt unwanted memories while leaving others intact. The number of the new grant is 1R01MH105400. Scripps Florida Scientists Win $2.2 Million to Expand Study of Innovative Obesity Therapy Scientists from the Florida campus of The Scripps Research Institute (TSRI) have been awarded nearly $2.2 million by the National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health (NIH) to advance an innovative approach to the treatment of obesity, a serious health problem that affects more than one-third of all Americans. Anutosh Chakraborty, a TSRI assistant professor, is the principal investigator of the new five-year project. Obesity, especially when combined with type 2 diabetes, leads to conditions including coronary heart disease, stroke, hypercholesterolemia, fatty liver, sleep apnea, osteoarthritis, certain cancers and various other diseases. If current trends continue, the number of Americans who are obese could reach 50 percent by 2030, according to the Trust for America's Health and the Robert Wood Johnson Foundation. According to Britain’s Fiscal Times, the estimated cost of obesity in the United States is already $305.1 billion annually. Current medications have limited success. In an effort to address this dilemma, scientists want to identify relevant proteins, especially enzymes, to target with new and more effective drug candidates. “Anti-obesity drugs generally work on reducing how much you eat or absorb,” Chakraborty said. “We investigate the problem from a different perspective.” Chakraborty and his colleagues discovered that an enzyme called inositol hexakisphosphate kinase-1 (IP6K1) plays a significant role in promoting the action of insulin on energy/fat storage. Mice without IP6K1 are not only lean on regular chow diet, they are also protected against high-fat-diet-induced obesity and insulin resistance. “IP6K1 knockout mice eat a similar amount of food, yet are lean as they efficiently expend the extra energy,” he said. “For us, that means that IP6K1 is the regulating factor when it comes to energy storage. Conversely, abnormal regulation of IP6K1 leads to obesity and insulin resistance. The new grant will allow us to identify the underlying mechanisms of how it works.”

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In addition to gaining a broader understanding of the fundamental mechanism by which IP6K1 regulates metabolism, Chakraborty and his colleagues—including Scripps Florida’s Ted Kamenecka, assistant professor and associate scientific director of the Translational Research Institute, and Michael Cameron, associate professor of molecular therapeutics and DMPK—are working on the development of drugs which are expected to treat obesity, type 2 diabetes and other metabolic diseases via IP6K1 inhibition. The number of the grant is 1R01DK103746-01A1. Scripps Florida Scientists Win $2.1 Million to Study Protein Linked to Parkinson’s Disease Scientists from the Florida campus of The Scripps Research Institute (TSRI) have been awarded $2.1 million from the National Institute of Neurological Disorders and Stroke of The National Institutes of Health (NIH) to study a protein that has been closely linked in animal models to Parkinson’s disease and Huntington’s disease. TSRI Assistant Professor Srinivasa Subramaniam will be the principal investigator of the new five-year grant. The focus of the new study is a multifunctional protein known as rapamycin (mTOR), which is involved in embryonic development, cancer and diabetes. Malfunction in mTOR activity—either too much or too little—has also been linked to a variety of brain dysfunctions such as epilepsy, mental retardation, Huntington’s disease and Parkinson’s disease. In the new project, the researchers will use a wide variety of techniques to examine the role and regulation of this protein in a brain region called the striatum, which controls motor, psychiatric and cognitive functions. “Even though mTOR is widely expressed throughout the body, its brain-specific regulation and function remain unclear,” Subramaniam said. “While we know that inhibiting mTOR protects against symptoms of Huntington’s and Parkinson’s diseases in animal models, the new grant will help us answer two critical questions: ‘How is mTOR regulated, and what happens when it is depleted selectively in the striatum?’ ” Subramaniam’s long-term goal is to understand the system well enough to advance new therapies. The number of the grant is 1R01NS087019. Scripps Florida Scientists Win $1.5 Million Grant to Develop New Cancer Drugs Scientists from the Florida campus of The Scripps Research Institute (TSRI) have been awarded a $1.5 million grant from the National Institutes of Health (NIH) to develop drug candidates that could treat cancer and neurodegenerative disease.

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Derek Duckett, a TSRI associate professor of molecular therapeutics, is the co-principal investigator for the three-year study, along with John Cleveland of the Moffitt Cancer Center in Tampa, Florida. Duckett, Cleveland and their teams will look for compounds that affect a key enzyme involved in the degradation and ultimate recycling of damaged cellular material. This process, called “autophagy,” is an ancient, cannibalistic (literally “self-eating”) pathway that acts as the main recycling center of all cells. In autophagy, bulk cytoplasmic material and aged or damaged organelles are recycled via the lysosome to recoup essential building blocks and adenosine triphosphate (ATP) as a survival strategy during times of stress or nutrient limitation. Autophagy is an important cell survival pathway, and any defects in its regulation can lead to a variety of disorders, including neurodegenerative disorders, liver disease and cancer. The study is focused on targeting a particular enzyme, UNC-51-like kinase-1 (Ulk1), a critical on-off switch that regulates this pathway. “Using these funds, we will identify new inhibitors of Ulk1,” Duckett said. “Developing selective molecular probes that function as Ulk1-specific inhibitors would improve our understanding of the autophagy pathway, its relationship to cancer and its utility as a target that could augment conventional or targeted anti-cancer treatments.” Duckett and his colleagues plan to use the high-throughput screening facilities at Scripps Florida and the Scripps Drug Discovery Library and its 650,000-plus library of small-molecule compounds. The number of the new grant from the NIH National Institute of General Medical Sciences is 1R01GM113972. Scripps Florida Scientists Awarded $1.2 Million to Find Drug Candidates that Could Treat a Wide Range of Cancers Scientists from the Florida campus of The Scripps Research Institute (TSRI) have been awarded $1.2 million from the National Cancer Institute of the National Institutes of Health (NIH) to accelerate the development of drug candidates to curb one of the most important drivers of human cancer. TSRI Associate Professors Joseph Kissil and Louis Scampavia will be co-principal investigators for the three-year grant, which will focus on the “Hippo-YAP signaling pathway.” “This pathway, which was discovered less than a decade ago, appears to regulate processes that are closely linked to an increasing number of cancers,” Kissil said. “The more we study it, the more we see its involvement. This new grant will help expand our investigation.” The Hippo-YAP signaling pathway has been found active in breast, colorectal and liver cancers, in hepatocellular and squamous cell carcinoma, and in melanoma of the eye. Cancers initiated through this pathway tend to thrive and proliferate, relatively immune to destruction from programmed cell death.

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Kissil, Scampavia and their colleagues plan to use Scripps Florida’s ultra-high-throughput screening resources and the campus’s library of more than 600,000 compounds to develop a series of screens to identify and optimize compounds to target the pathway and combat cancer. The number of the grant is 1R01CA184277. PART B: SCIENTIFIC ACCOMPLISHMENTS Scripps Florida Scientists Uncover New Compounds that Could Affect Circadian Rhythm Scientists from the Florida campus of The Scripps Research Institute (TSRI) have discovered a surprising new role for a pair of compounds—which have the potential to alter circadian rhythm, the complex physiological process that responds to a 24-hour cycle of light and dark and is present in most living things. At least one of these compounds could be developed as a chemical probe to uncover new therapeutic approaches to a range of disorders, including diabetes and obesity. The study, which was published online ahead of print by the Journal of Biological Chemistry, focuses on a group of proteins known as REV-ERBs, a superfamily that plays an important role in the regulation of circadian physiology, metabolism and immune function. The new study shows that the two compounds, cobalt protoporphyrin IX (CoPP) and zinc protoporphyrin IX (ZnPP), bind directly to REV-ERBs. REV-ERBs are normally regulated by heme, a molecule that binds to hemoglobin, helps transport oxygen from the bloodstream to cells and plays a role in producing cellular energy. While heme activates REV-ERB, CoPP and ZnPP inhibit it. “These compounds are like heme, but when you swap out their metal centers their functions are different,” said Doug Kojetin, a TSRI associate professor who led the study. “This makes us think that the key is the chemistry of the metal ion itself. Altering the chemistry of this metal center may be an opportune way to target REV-ERB for diabetes and obesity.” Kojetin and his colleagues recently demonstrated that synthetic REV-ERB agonists, like the new compounds, reduce body weight in mice that were obese due to diet. The first authors of the study, “Structure of REV-ERB_ Ligand-binding Domain Bound to a Porphyrin Antagonist,” are Edna Matta-Camacho of McGill University, Montreal and Subhashis Banerjee of the University of Texas Southwestern Medical Center. Other authors of the study include Travis S. Hughes and Laura A. Solt of TSRI; and Yongjun Wang and Thomas P. Burris of St. Louis University School of Medicine.

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The work was supported by the National Institutes of Health (grants DK080201 and DK101871), the James and Esther King Biomedical Research Program from the Florida Department of Health (grant 1KN-09) and the State of Florida. The work can be accessed at http://www.jbc.org/content/early/2014/05/28/jbc.M113.545111.full.pdf+html?sid=bef6d5e6-a7dd-4ca5-9f57-6fba6122a6b5 Scripps Florida Scientists Shed New Light on Nerve Cell Growth Amidst the astounding complexity of the billions of nerve cells and trillions of synaptic connections in the brain, how do nerve cells decide how far to grow or how many connections to build? How do they coordinate these events within the developing brain? In a new study, scientists from the Florida campus of The Scripps Research Institute (TSRI) have shed new light on these complex processes, showing that a particular protein plays a far more sophisticated role in neuron development than previously thought. The study, published in the journal PLOS Genetics, focuses on the large, intracellular signaling protein RPM-1 that is expressed in the nervous system. TSRI Assistant Professor Brock Grill and his team show the surprising degree to which RPM-1 harnesses sophisticated mechanisms to regulate neuron development. Specifically, the research sheds light on the role of RPM-1 in the development of axons or nerve fibers—the elongated projections of nerve cells that transmit electrical impulses away from the neuron via synapses. Some axons are quite long; in the sciatic nerve, axons run from the base of the spine to the big toe. “Collectively, our recent work offers significant evidence that RPM-1 coordinates how long an axon grows with construction of synaptic connections,” said Grill. “Understanding how these two developmental processes are coordinated at the molecular level is extremely challenging. We’ve now made significant progress.” Putting Together the Pieces The study describes how RPM-1 regulates the activity of a single protein known as DLK-1, a protein that regulates neuron development and plays an essential role in axon regeneration. RPM-1 uses PPM-2, an enzyme that removes a phosphate group from a protein thereby altering its function, in combination with intrinsic ubiquitin ligase activity to directly inhibit DLK-1. “Studies on RPM-1 have been critical to understanding how this conserved family of proteins works,” said Scott T. Baker, the first author of the study and a member of Grill’s research team. “Because RPM-1 plays multiple roles during neuronal development, you wouldn’t want to interfere with it. But exploring the role of PPM-2 in controlling DLK-1 and axon regeneration could be worthwhile—and could have implications in neurodegenerative diseases.”

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The Grill lab has also explored other aspects of how RPM-1 regulates neuron development. A related study, also published in PLOS Genetics, shows that RPM-1 functions as a part of a novel pathway to control β-catenin activity—this is the first evidence that RPM-1 works in connection with extracellular signals, such as a family of protein growth factors known as Wnts, and is part of larger signaling networks that regulate development. A paper in the journal Neural Development shows that RPM-1 is localized at both the synapse and the mature axon tip, evidence that RPM-1 is positioned to potentially coordinate the construction of synapses with regulation of axon extension and termination. In addition to Grill and Baker, Erik Tulgren of the University of Minnesota, Willy Bienvenut of the Campus de Recherche de Gif, France, as well as Karla Opperman and Shane Turgeon of TSRI contributed to the study entitled, “RPM-1 Uses Both Ubiquitin Ligase and Phosphatase-Based Mechanisms to Regulate DLK-1 during Neuronal Development.” For more information, see http://www.plosgenetics.org/article/info%3Adoi%2F10.1371%2Fjournal.pgen.1004297 The first author of the study, “The Nesprin Family Member ANC-1 Regulates Synapse Formation and Axon Termination by Functioning in a Pathway with RPM-1 and β-Catenin,” is Erik Tulgren of the University of Minnesota. Other authors include Shane Turgeon and Karla Opperman of TSRI. For more information, see http://www.plosgenetics.org/doi/pgen.1004481 The first author of the study, “RPM-1 Is Localized to Distinct Subcellular Compartments and Regulates Axon Length in GABAergic Motor Neurons,” is Karla Opperman of TSRI. For more information, see http://www.neuraldevelopment.com/content/9/1/10 The work was supported by the National Institutes of Health (grant R01 NS072129) and the National Science Foundation (grant IOS- 1121095). Scripps Florida Scientists Identify Gene that Plays a Surprising Role in Combating Aging It is something of an eternal question: Can we slow or even reverse the aging process? Even though genetic manipulations can, in fact, alter some cellular dynamics, little is known about the mechanisms of the aging process in living organisms. Now scientists from the Florida campus of The Scripps Research Institute (TSRI) have found in animal models that a single gene plays a surprising role in aging that can be detected early on in development, a discovery that could point toward the possibility of one day using therapeutics, even some commonly used ones, to manipulate the aging process itself. “We believe that a previously uncharacterized developmental gene known as Spns1 may mediate the aging process,” said Shuji Kishi, a TSRI assistant professor who led the study, published recently by the journal PLOS Genetics. “Even a partial loss of Spns1 function can speed aging.” Using various genetic approaches to disturb Spns1 during the embryonic and/or larval stages of zebrafish—which have emerged as a powerful system to study diseases associated with development

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and aging—the scientists were able to produce some models with a shortened life span, others that lived long lives. While most studies of “senescence”—declines in a cell's power of division and growth—have focused on later stages of life, the study is intriguing in exploring this phenomenon in early stages. “Mutations to Spns1 both disturbs developmental senescence and badly affects the long-term bio-chronological aging process,” Kishi said. The new study shows that Spns1, in conjunction with another pair of tumor suppressor genes, beclin 1 and p53 can, influences developmental senescence through two differential mechanisms: the Spns1 defect was enhanced by Beclin 1 but suppressed by ‘basal’ p53. In addition to affecting senescence, Spns1 impedes autophagy, the process whereby cells remove unwanted or destructive proteins and balance energy needs during various life stages. Building on their insights from the study, Kishi and his colleagues noted in the future therapeutics might be able influence aging through Spns1. He noted one commonly used antacid, Prilosec, has been shown to temporarily suppress autophagic abnormality and senescence observed in the Spns1 deficiency. The first author of the study, “Aberrant Autolysosomal Regulation Is Linked to The Induction of Embryonic Senescence: Differential Roles of Beclin 1 and p53 in Vertebrate Spns1 Deficiency,” is Tomoyuki Sasaki of TSRI. Other authors include Shanshan Lian, Jie Qi, Sujay Guha, Jennifer L. Johnson, Sergio D. Catz and Matthew Gill of TSRI; Peter E. Bayliss of the University Health Network, Toronto, Canada; Christopher E. Carr of the Massachusetts Institute of Technology; Patrick Kobler and Kailiang Jia of Florida Atlantic University; and Daniel J. Klionsky of the University of Michigan. The paper can be accessed at http://www.plosgenetics.org/article/info%3Adoi%2F10.1371%2Fjournal.pgen.1004409 The work was supported by The Ellison Medical Foundation, Glenn Foundation for Medical Research, A-T Children's Project), the National Institutes of Health (NIH) National Institute of Aging (AG022641) and the National Institute of General Medical Sciences (GM053396, GM101508). Scientists Find Ancient Protein-Building Enzymes Have Undergone Metamorphosis and Evolved Diverse New Functions The Previously Unrecognized Layer of Biology Could Offer New Drug Targets Scientists at The Scripps Research Institute (TSRI) and Hong Kong University of Science and Technology (HKUST) and their collaborators have found that ancient enzymes, known for their fundamental role in translating genetic information into proteins, evolved myriad other functions in humans. The surprising discovery highlights an intriguing oddity of protein evolution as well as a potentially valuable new class of therapeutic proteins and therapeutic targets.

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“These new protein variants represent a previously unrecognized layer of biology—the ramifications of this discovery are now unfolding,” said team leader Paul Schimmel, Ernest and Jean Hahn Professor of Molecular Biology and Chemistry at TSRI (California and Florida) who also holds an appointment at HKUST Jockey Club Institute for Advanced Study (IAS). Mingjie Zhang, a co-author of the paper who is IAS Senior Fellow and Kerry Holdings Professor of Science, Division of Life Science at HKUST, added, “This breakthrough finding not only uncovers a vast area of new biology, but also provides opportunities to develop protein-based drugs for various human diseases associated with malfunctions of these newly discovered proteins. The successful collaboration among scientists from HKUST, TSRI and two biotech companies from the U.S. and Hong Kong also serves as a wonderful example of the close connections between basic research and biotechnology development.” The findings appear in the July 18, 2014 issue of Science. Greater Complexity The discovery concerns aminoacyl tRNA synthetases (AARSs), a group of 20 enzymes whose most basic function is to connect the nucleotide codes contained in genes to their corresponding protein building-blocks, namely, the 20 amino acids. Because AARSs are essential for the translation of genetic information into working proteins, they are found in all life forms on the planet. Scientists have been finding evidence in recent years that AARS enzymes exist in greater complexity in more evolutionarily advanced organisms. In essence, these enzymes have acquired new segments or “domains.” Curiously, these domains have no apparent relevance to protein translation. While largely absent in lower forms of life, these domains are added in a progressive and accretive way in evolution, in the long ascent over billions of years to humans. To investigate further, Schimmel collaborated with colleagues at TSRI in California and Florida, HKUST (including at the IAS), the San Diego biotech company aTyr Pharma (which Schimmel co-founded), Stanford and the Hong Kong biotech company Pangu Biopharma (an aTyr subsidiary). Using advanced, sensitive techniques, the team identified nearly 250 previously unknown gene-transcript variants of AARS in different human cell types. These variants, known as splice variants, are alternative assemblies of the discrete sequences of information (exons) contained in AARS genes. Genes frequently contain multiple exons that can be spliced together in alternative ways—thus in principle enabling a single active gene to encode multiple proteins with different functions. But the new findings suggest that evolution has been unusually prolific at creating AARS splice variants. Potential New Class of Drug Targets The team’s further investigations revealed that the new AARS variants often are produced only in specific cell types such as brain or immune cells and/or appear only during certain stages of development.

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Remarkably, most of the new splice variants lack entirely the standard “catalytic domain” that supports protein translation—confirming that they are not directly involved in that fundamental process. Initial screening of the biological activities of these variants hinted at a diversity of other functions. One variant selected for analysis turned out to be a powerful driver for the proliferation of muscle fiber cells in a laboratory dish. Schimmel and his collaborators will now turn to more comprehensive studies of the new AARS variants and their specific functions. “We believe that these proteins have relevance to multiple human diseases,” he said. “They thus represent a very important class of new protein therapeutics analogous to widely used injectable protein therapeutics such as growth hormone, insulin, erythropoietin (EPO) (which regulates red blood cell production) and granulocyte colony-stimulating factor (G-CSF) (which stimulates the bone marrow).” The first author of the report, “Human tRNA synthetase catalytic nulls with diverse functions,” was Wing-Sze Lo from the IAS-Scripps R&D Laboratory at HKUST and Pangu Biopharma. In addition to Schimmel and Zhang, other authors included Zhiwen Xu, Ching-Fun Lau, Feng Wang and Jie Zhou, also from the IAS-Scripps R&D Laboratory at HKUST and Pangu Biopharma; John D. Mendlein, Leslie A. Nangle and Kyle P. Chiang of aTyr Pharma; Kin-Fai Au, formerly at Stanford University and now at the University of Iowa; Wing Hung Wong of Stanford University; Min Guo of TSRI’s Florida campus; and Elisabeth Gardiner and Xiang-Lei Yang from TSRI’s California campus. Funding for the research was provided by the Hong Kong Government’s Innovation and Technology Fund (grants UIM181, UIM192 and UIM199), the National Foundation for Cancer Research, the National Institutes of Health (grants R01CA92577, R01GM088278, R01NS085092, R01HG005717 and R01GM100136), aTyr Pharma and Pangu Biopharma. Scripps Florida Scientists Find Genetic Mutations Linked to Salivary Gland Tumors The Findings May Point the Way to New Cancer Treatments Research conducted at the Florida campus of The Scripps Research Institute (TSRI) has discovered links between a set of genes known to promote tumor growth and mucoepidermoid carcinoma, an oral cancer that affects the salivary glands. The discovery could help physicians develop new treatments that target the cancer’s underlying genetic causes. The research, published recently online ahead of print by the Proceedings of the National Academy of Sciences, shows that a pair of proteins joined together by a genetic mutation—known as CRTC1/MAML2 (C1/M2)—work with MYC, a protein commonly associated with other cancers, to promote the oral cancer’s growth and spread. “This research provides new insights into the molecular mechanisms of these malignances and points to a new direction for potential therapies,” says TSRI biologist Michael Conkright, Ph.D., who led the study.

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The C1/M2 protein is created when the genes encoding CRTC1 and MAML2 mutate into a single gene through a process known as chromosomal translocation. Such mutant “chimera” genes are linked to the formation of several forms of cancer. The team discovered that the C1/M2 protein further activates genetic pathways regulated by MYC, in addition to CREB, to begin a series of cellular changes leading to the development of mucoepidermoid carcinoma. “The identification of unique interactions between C1/M2 and MYC suggests that drugs capable of disrupting these interactions may have therapeutic potential in the treatment of mucoepidermoid carcinomas,” said Antonio L. Amelio, Ph.D., first author of the study who is now assistant professor with the UNC School of Dentistry and member of the UNC Lineberger Comprehensive Cancer Center. Researchers have known about the role of C1/M2 and its interactions with another protein, CREB, in the development of mucoepidermoid carcinoma, and physicians screen patients for the presence of the C1/M2 protein when testing for this cancer. These new findings deepen the understanding of C1/M2’s role by revealing that it works with a family of cancer-associated genes known as the MYC family to drive the cellular changes necessary for a tumor to develop. The discovery of these new protein interactions may also reveal insights into the mechanisms behind other cancers that arise due to other genetic mutations involving the CREB and MYC pathways. In addition to Conkright and Amelio, other authors of the study, “CRTC1/MAML2 gain-of-function interactions with MYC create a gene signature predictive of cancers with CREB–MYC involvement,” include Mohammad Fallahi of IT Informatics, Franz X. Schaub, Mariam B. Lawani, Adam S. Alperstein, Mark R. Southern, Brandon M. Young and John L. Cleveland of TSRI, and Min Zhang, Lizi Wu, Maria Zajac-Kaye and Frederic J. Kaye of Shands Cancer Center, University of Florida (Gainesville). The research was supported in part by a Howard Temin Pathway to Independence Award in Cancer Research from the National Cancer Institute (NCI) (K99-CA157954), National Institutes of Health/NCI R01 Grant CA100603, a PGA National WCAD Cancer Research Fellowship and Ruth L. Kirschstein National Research Service Award from the National Cancer Institute (F32-CA134121), the Margaret Q. Landerberger Research Foundation, a Swiss National Foundation Postdoctoral Fellowship and monies from the State of Florida to TSRI’s Scripps Florida. A Gene Linked to Disease Found to Play a Critical Role in Normal Memory Development It has been more than 20 years since scientists discovered that mutations in the gene huntingtin cause the devastating progressive neurological condition Huntington’s disease, which involves involuntary movements, emotional disturbance and cognitive impairment. Surprisingly little, however, has been known about the gene’s role in normal brain activity. Now, a study from The Scripps Research Institute’s (TSRI’s) Florida campus and Columbia University shows it plays a critical role in long-term memory.

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“We found that huntingtin expression levels are necessary for what is known as long-term synaptic plasticity—the ability of the synapses to grow and change—which is critical to the formation of long-term memory,” said TSRI Assistant Professor Sathyanarayanan V. Puthanveettil, who led the study with Nobel laureate Eric Kandel of Columbia University. In the study, published recently by the journal PLOS ONE, the team identified an equivalent of the human huntingtin protein in the marine snail Aplysia, a widely used animal model in genetic studies, and found that, just like its human counterpart, the protein in Aplysia is widely expressed in neurons throughout the central nervous system. Using cellular models, the scientists studied what is known as the sensory-to-motor neuron synapse of Aplysia—in this case, gill withdrawal, a defensive move that occurs when the animal is disturbed. The study found that the expression of messenger RNAs of huntingtin—messenger RNAs are used to produce proteins from instructions coded in genes—is increased by serotonin, a neurotransmitter released during learning in Aplysia. After knocking down production of the huntingtin protein, neurons failed to function normally. “During the learning, production of the huntingtin mRNAs is increased both in pre- and post-synaptic neurons—that is a new finding,” Puthanveettil said. “And if you block production of the protein either in pre- or post-synaptic neuron, you block formation of memory.” The findings could have implications for the development of future treatments of Huntington’s disease. While the full biological functions of the huntingtin protein are not yet fully understood, the results caution against a therapeutic approach that attempts to eliminate the protein entirely. The first author of the study, “Huntingtin Is Critical Both Pre- and Postsynaptically for Long-Term Learning-Related Synaptic Plasticity in Aplysia,” is Yun-Beom Choi of Columbia University. Other authors include Beena M. Kadakkuzha, Xin-An Liu and Komolitdin Akhmedov of TSRI. For more information on the study, see http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0103004 The work was supported by the Howard Hughes Medical Institute, the National Institutes of Health (Grant NS053415), the Whitehall Foundation and the State of Florida. Scripps Florida and Mayo Clinic Team Up to Successfully Target Common Mutation in Lou Gehrig’s Disease and Frontotemporal Dementia An international team led by scientists from the Florida campuses of The Scripps Research Institute (TSRI) and the Mayo Clinic have for the first time successfully designed a therapeutic strategy targeting a specific genetic mutation that causes a common form of amyotrophic lateral sclerosis (ALS), better known as Lou Gehrig’s disease, as well a type of frontotemporal dementia (FTD). The scientists developed small-molecule drug candidates and showed they interfere with the synthesis of an abnormal protein that plays a key role in causing both diseases. The team also developed biomarkers that can test the efficacy of this and other therapies.

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The study, led by Professor Matthew Disney of TSRI and Professor of Neuroscience Leonard Petrucelli of the Mayo Clinic, was published online ahead of print August 14, 2014 in the journal Neuron. “Our small molecules target a genetic defect that is by far the most major cause of familial ALS, and if you have this defect you are assured of getting ALS or FTD,” Disney said. “Our findings show for the first time that targeting this mutation with a small-molecule drug candidate can inhibit toxic protein translation—and establishes that it could be possible to treat a large number of these patients, but this is just the start of these studies and additional investigations need to be done.” Currently, ALS is usually fatal two to five years after diagnosis, and there is no effective treatment for FTD, a neurodegenerative disease that destroys neurons in the frontal lobes of the brain. Toxic Buildup The mutation that can cause both diseases affects a gene known as C90RF72 and involves a repeat expansion, a longer than usual repetitive genetic sequence. This results in abnormal strands of RNA and the production of toxic “c9RAN proteins.” Disney and his Scripps Florida colleagues initially designed three small-molecule drug candidates that decreased RNA translation or production of these toxic proteins in cell culture. The Mayo team developed the patient-derived cell models in which to test the compounds and the biomarker to assess compound activity. Both teams then worked together to show that the lead agent’s mode of action was targeting the toxic RNA, binding to and blocking the toxic RNA’s ability to interact with other key proteins. Two of the compounds significantly decreased levels of the toxic protein. Using a series of increasing dosages of the drug candidates, the scientists found that the highest dosage of one reduced the toxic protein by nearly 50 percent. The scientists also discovered that c9RAN proteins produced by the abnormal RNA can be measured in the spinal fluid of ALS patients. They are now evaluating whether these proteins are also present in spinal fluid of patients diagnosed with FTD. “A decrease in the levels of toxic proteins in cerebrospinal fluid in response to treatment would demonstrate the drug is working,” Petrucelli said. “While additional studies must be done, this finding suggests that these proteins may provide a direct means to measure a patient’s response to experimental drugs that target abnormal RNA.” Toxic proteins found in spinal fluid could also become an enrollment tool in human clinical trials, added Disney, who was enthusiastic about the collaboration with the Mayo Clinic and the larger team. “Our collective biological and chemical expertise made this research possible,” he said. “This is just the beginning of what we can do together.”

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The first authors of the study, “Discovery of a Biomarker and Lead Small Molecules to Target r(GGGGCC)-Associated Defects in c9FTD/ALS,” are Zhaoming Su of TSRI and Yongjie Zhang, Tania F. Gendron and Peter O. Bauer of the Mayo Clinic. Other authors include Wang-Yong Yang and Erik Fostvedt of TSRI; Jeannie Chew, Karen Jansen-West, Veronique V. Belzil, Pamela Desaro, Amelia Johnston, Karen Overstreet, Bradley F. Boeve, Dennis Dickson, Rosa Rademakers and Kevin B. Boylan of the Mayo Clinic; Mary Kay Floeter of the National Institute of Neurological Disorders and Stroke; Bryan J. Traynor of the National Institute on Aging; Claudia Morelli of the IRCCS Instituto Auxologico Italiano, Milan, Italy; Antonia Ratti and Vincenzo Silani of the IRCCS Instituto Auxologico Italiano, Milan, Italy and the Universita degli Studi di Milano, Milan, Italy; and Robert H. Brown and Jeffrey D. Rothstein of Johns Hopkins University. The work was supported the National Institute on Aging of the National Institutes of Health (grants R01GM097455, R01AG026251 and P50AG016574); National Institute of Neurological Disorders and Stroke (grants R21NS074121, R21NS079807, R21NS084528, R01NS088689, R01NS063964, R01NS077402 and R01NS050557); the American Recovery and Reinvestment Act of 2009 (awards RC2-NS070-342 and P01NS084974); National Institute of Environmental Health Services (grant R01ES20395); Department of Defense (ALSRP AL130125); Mayo Clinic Foundation; Mayo Clinic Center for Regenerative Medicine; Mayo Clinic Center for Individualized Medicine; ALS Association; Alzheimer's Association; Robert Packard Center for ALS Research at Johns Hopkins; Target ALS; Project ALS; Angel Fund; the Italian Ministry of Health (RF-2009-1473856) and the European Commission. Scripps Florida Scientists Make Diseased Cells Synthesize Their Own Drug In a new study that could ultimately lead to many new medicines, scientists from the Florida campus of The Scripps Research Institute (TSRI) have adapted a chemical approach to turn diseased cells into unique manufacturing sites for molecules that can treat a form of muscular dystrophy. “We’re using a cell as a reaction vessel and a disease-causing defect as a catalyst to synthesize a treatment in a diseased cell,” said TSRI Professor Matthew Disney. “Because the treatment is synthesized only in diseased cells, the compounds could provide highly specific therapeutics that only act when a disease is present. This means we can potentially treat a host of conditions in a very selective and precise manner in totally unprecedented ways.” The promising research was published recently in the international chemistry journal Angewandte Chemie. Targeting RNA Repeats In general, small, low molecular weight compounds can pass the blood-brain barrier, while larger, higher weight compounds tend to be more potent. In the new study, however, small molecules became powerful inhibitors when they bound to targets in cells expressing an RNA defect, such as those found in myotonic dystrophy.

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Myotonic dystrophy type 2, a relatively mild and uncommon form of the progressive muscle weakening disease, is caused by a type of RNA defect known as a “tetranucleotide repeat,” in which a series of four nucleotides is repeated more times than normal in an individual’s genetic code. In this case, a cytosine-cytosine-uracil-guanine (CCUG) repeat binds to the protein MBNL1, rendering it inactive and resulting in RNA splicing abnormalities that, in turn, results in the disease. In the study, a pair of small molecule “modules” the scientists developed binds to adjacent parts of the defect in a living cell, bringing these groups close together. Under these conditions, the adjacent parts reach out to one another and, as Disney describes it, permanently hold hands. Once that connection is made, the small molecule binds tightly to the defect, potently reversing disease defects on a molecular level. “When these compounds assemble in the cell, they are 1,000 times more potent than the small molecule itself and 100 times more potent than our most active lead compound,” said Research Associate Suzanne Rzuczek, the first author of the study. “This is the first time this has been validated in live cells.” Click Chemistry Construction The basic process used by Disney and his colleagues is known as “click chemistry”—a process invented by Nobel laureate K. Barry Sharpless, a chemist at TSRI, to quickly produce substances by attaching small units or modules together in much the same way this occurs naturally. “In my opinion, this is one unique and a nearly ideal application of the process Sharpless and his colleagues first developed,” Disney said. Given the predictability of the process and the nearly endless combinations, translating such an approach to cellular systems could be enormously productive, Disney said. RNAs make ideal targets because they are modular, just like the compounds for which they provide a molecular template. Not only that, he added, but many similar RNAs cause a host of incurable diseases such as ALS (Lou Gehrig’s Disease), Huntington’s disease and more than 20 others for which there are no known cures, making this approach a potential route to develop lead therapeutics to this large class of debilitating diseases. In addition to Rzuczek and Disney, the other author of the study, “A Toxic RNA Catalyzes the In Cellulo Synthesis of Its Own Inhibitor,” is HaJeung Park of TSRI. For more information on the study, see http://onlinelibrary.wiley.com/doi/10.1002/anie.201406465/abstract The work was supported by the Muscular Dystrophy Foundation, the Myotonic Dystrophy Foundation and the State of Florida.

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Team Identifies Important Regulators of Immune Cell Response In a collaborative study, scientists from the Florida campus of The Scripps Research Institute (TSRI) and the La Jolla Institute for Allergy and Immunology have developed a more effective method to determine how immune cells called T cells differentiate into specialized types of cells that help eradicate infected cells and assist other immune cells during infection. The new approach, published recently by the journal Immunity, could help accelerate laboratory research and the development of potential therapeutics, including vaccines. The method may also be used to identify the genes that underlie tumor cell development. There are approximately 40,000 genes in each of our cells, but functions for only about half of them are known. The classical approach to determine the function of individual genes is slow. “Typically, studies to identify differentiation players are done one gene at a time,” said Associate Professor Matthew Pipkin of TSRI, who led the study with Professor Shane Crotty of the La Jolla Institute for Allergy and Immunology. “Our study describes a novel method that can ‘screen’ entire gene families to discover the functions of a large number of individual genes simultaneously, a far more efficient methodology.” In the new study, the team examined genes that regulate the specialization of T cells into “effector” cells that eliminate pathogens during infection and “memory” cells that survive long-term to maintain guard after the first infection has been cleared, keeping the same pathogens from re-infecting the body after it has fought them off once. In their experiments, Pipkin, Crotty and their colleagues created a mixture of T cells, identical except that the expression of a different gene was interrupted in each cell so the pool of cells represented disruption of a large set of genes. The researchers then assessed the cells’ response to lymphocytic choriomeningitis virus (LCMV). Before-and-after-infection studies revealed which cells with interrupted genes had emerged after infection; cells in which disruption of a particular gene resulted in it being lost from the mixture indicated the gene played a role in promoting the cell’s development into an antiviral T cell. The study successfully identified two previously unknown factors that work together during T cell differentiation—Cyclin T1 and its catalytic partner Cdk9, which together form the transcription elongation factor (P-TEFb). While widely expressed throughout the body and used in a number of developmental processes, the factors were previously unknown to be important in the differentiation of both antiviral CD4 and CD8 T cells. “One of the regulators we uncovered normally enhances effector T cell differentiation at the expense of generating memory T cells and T cells that orchestrate antibody production,” Pipkin said. “That’s one candidate that you’d want to ‘turn down’ if you wanted to create more T cells that form memory cells and promote a more effective antibody response—something that would be extremely helpful in developing a vaccine.”

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The first authors of the study, “In Vivo RNA Interference Screens Identify Regulators of Antiviral CD4+ and CD8+ T Cell Differentiation,” are Runqiang Chen and Simon Bélanger of the La Jolla Institute for Allergy and Immunology. Other authors include Megan A. Frederick of TSRI; and Bin Li, Robert J. Johnston, Nengming Xiao, Yun-Cai Liu, Sonia Sharma, Bjoern Peters and Anjana Rao of the La Jolla Institute for Allergy and Immunology. See http://www.cell.com/immunity/abstract/S1074-7613(14)00272-6 This work was supported by the National Institutes of Health (RC4 AI092763, R01 AI095634, R01 CA42471, R01 072543 and U19 AI109976) and Frenchman’s Creek Women for Cancer Research. New Clinical Trial Data: Scripps Research Institute MS Drug Candidate Also Shows Promise for Ulcerative Colitis Positive new clinical data were released recently on a drug candidate for ulcerative colitis that was first discovered and synthesized at The Scripps Research Institute (TSRI). According to recent results from a Phase 2 study of 199 patients with active, moderate to severe disease, the drug candidate RPC1063 has potential to significantly improve the treatment paradigm for ulcerative colitis patients. The latest results show that, after eight weeks of treatment with a 1 mg dose of RPC1063, 16.4 percent of patents were in clinical remission, as compared to 6.2 percent of patients on placebo. “We are delighted that RPC1063 is showing promise for ulcerative colitis patients in addition to its already significant efficacy and safety data in multiple sclerosis,” said TSRI Professor Hugh Rosen, who together with Professor Ed Roberts led the team that discovered RPC-1063. “Research carried out at TSRI since 2002 has led to the discovery of fundamental mechanisms that can be modulated for potential treatments of a variety of autoimmune diseases including ulcerative colitis and multiple sclerosis, and the unique multidisciplinary environment in chemistry and biology at TSRI allowed this progression to clinical trials.” The clinical trial, sponsored by Receptos, Inc., the San Diego biotechnology company now developing the drug, also showed that RPC1063 was generally well tolerated. Ulcerative colitis is a chronic condition that involves inflammation and sores in the inner lining of the digestive tract. Ulcerative colitis is an inflammatory bowel disease, which, along with Crohn’s disease, affects more than one million people nationwide, according to the U.S. Centers for Disease Control and Prevention. Some people have mild disease, while others are affected with life-threatening complications. While existing medications for ulcerative colitis do help some patients, 23 to 45 percent of ulcerative colitis sufferers progress and eventually require surgical removal of all or part of the colon, according to the Crohn’s and Colitis Foundation of America. The drug candidate RPC1063 was derived from a screening “hit” from the National Institutes of Health molecular library at Scripps Florida’s Molecular Screening Center, using assay technology

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from the Rosen lab in La Jolla. The Roberts and Rosen labs then developed significant medicinal chemistry to turn that hit into a validated lead, and then ultimately a drug candidate. TSRI then licensed the compound to Receptos, which is developing RPC1063 for approval by the U.S. Food and Drug Administration. The latest results come from Receptos’s multi-national, multi-center, double-blind, randomized, placebo-controlled study investigating the effect of two active doses of RPC1063 versus placebo for the treatment of moderately to severely active ulcerative colitis. For more information on the results, see the press release from Receptos at http://ir.receptos.com/releasedetail.cfm?ReleaseID=878411 In light of the current positive results, Receptos plans to initiate a Phase 3 trial of RPC1063 for ulcerative colitis, as well as a Phase 2 study of the drug candidate for Crohn’s disease. The mechanism of RPC1063 (Sphingosine 1-Phosphate Receptor modulation) may also be significant in the treatment of other autoimmune diseases. Receptos is also currently evaluating the drug candidate in a Phase 3 study for the treatment of multiple sclerosis. Scripps Florida Scientists Uncover Major Factor in Development of Huntington’s Disease Scientists from the Florida campus of The Scripps Research Institute (TSRI) have uncovered a major contributor to Huntington’s disease, a devastating progressive neurological condition that produces involuntary movements, emotional disturbance and cognitive impairment. Using an animal model of Huntington’s disease, the new study shows that signaling by a specific protein can trigger onset of the disease and lead to exacerbation of symptoms. These findings, published in the October 28, 2014 issue of the journal Science Signaling, offer a novel target for drug development. It has been more than 20 years since scientists discovered that mutations in the gene huntingtin cause Huntington’s disease; the product of the gene, Huntingtin protein, is widely expressed is almost all of the cells in the body. The disease results in an early loss of neurons in the striatum, part of the forebrain that is responsible for coordinating thought with movement—when you want to move your arm, the striatum lets your muscles know. Unfortunately, the precise physiological role for huntingtin in disease onset and progression remains unclear. The new study, however, shows for the first time a functional connection between huntingtin and mTOR, a developmentally important gene that integrates signals from multiple pathways, such as growth factors and hormones, to regulate a variety of critical cell functions. Specifically, the scientists found that the huntingtin protein activates signaling by a protein complex known as mTORC1 (mechanistic-target of rapamycin kinase (mTOR) complex 1). Depleting huntingtin reduces mTORC1 activity; an overexpression of huntingtin increases it.

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“In our previous work, we showed that there is a protein in the striatum that interacts with huntingtin and makes it more toxic—this protein can activate mTORC1,” said Srinivasa Subramaniam, a TSRI biologist who led the study. “What we didn’t know was how TORC1 and huntingtin were related. What we found for the first time in this new study is that huntingtin can activate mTORC1 and increase its activity in the striatum of mice—thus prematurely initiating the disease.” In the new research, Subramaniam and his colleagues selectively deleted a gene that inhibits mTORC1 activity in the animal model striatum, which caused a relatively rapid increase in the severity of behavioral abnormalities related to Huntington’s disease, as well as premature death. “This indicates for the first time that huntingtin is a novel regulator of mTORC1 activity that contributes to the pathogenesis of the disease, at least in animal models,” he said. The researchers will continue to investigate the role of mTORC1 in Huntington’s and other age-dependent neurodegenerative diseases. “We think that huntingtin may regulate mTORC1 both in the brain and in other tissue,” said William Pryor, the first author of the study and a member of Subramaniam’s laboratory. “Our suspicion is that this exacerbation of mTORC1 might compromise autophagy—the pathway that recycles proteins and organelles—which has been implicated in neurodegeneration.” “Reducing mTORC1 activation either through drugs or low-protein foods may have a positive influence on preventing the disease process,” said Subramaniam. In addition to Subramaniam and Pryor, other authors of the study, “Huntingtin Regulates mTORC1 Pathway that Exacerbates Huntington Disease Pathogenesis,” include Neelam Shahani, Supriya Swarnkar, Wen-Chin Huang and Damon T. Page of TSRI; and Marta Biagioli and Marcy E. MacDonald of the Center for Human Genetic Research, Massachusetts General Hospital. The study was supported by the state of Florida and the O'Keeffe Neuroscience Scholar Award. Scripps Florida Scientists Unveil New Targets and Test to Develop Treatments for Memory Disorders In a pair of related studies, scientists from the Florida campus of The Scripps Research Institute (TSRI) have identified a number of new therapeutic targets for memory disorders and have developed a new screening test to uncover compounds that may one day work against those disorders. The two studies, one published in the journal Proceedings of the National Academy of Sciences (PNAS), the other in the journal ASSAY and Drug Development Technologies, could lead new approaches to some of the most problematic diseases facing a rapidly aging world population, including Alzheimer’s and Huntington’s diseases and dementia. “We are actively looking at molecules critical to memory formation, so these two studies work in parallel,” said Sathyanarayanan V. Puthanveettil, a TSRI biologist who led both studies. “In one

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study, we’re reaching for a basic understanding of the process, and in the other, we’re finding new ways to identify drug candidates so that we can cure these diseases.” Unlocking the ‘Synaptic Proteome’ The PNAS study is one of the first detailed descriptions of the proteins that are transported to the synapses, which as a group are called the “synaptic proteome.” Synapses are the part of a nerve cell (neuron) that passes electrochemical signals to other cells during functions such as memory storage. This new approach has the potential to advance our understanding of how synapses work, how their composition changes with learning and how brain diseases might affect them. “We know these molecules function in the synapse, and if we can regulate their function there may be some very good therapeutic opportunities there,” Puthanveettil said. The study focuses on kinesin, a molecular motor protein that plays a role in the transport of other proteins throughout a cell. Analyzing three kinesin complexes, the researchers found that approximately 40 to 50 percent of the protein cargos were synaptic proteins—and that the identity and location of these kinesins determine which proteins they transport. These results reveal a previously underappreciated role of kinesins in regulating the composition of the entire synaptic proteome. Interestingly, a bioinformatics analysis revealed the three kinesin cargo complexes examined in the study are involved in neurologic diseases. Approximately 60 cargos (out of 155) of the kinesin Kif5C are implicated in psychiatric disorders, while around 20 cargos of another kinesin Kif3A are implicated in developmental disorders. “This shows for the first time how kinesins expressed in the same neurons can carry substantially different cargos,” said Research Associate Xin-An Liu, the first author of the study. “We can use this approach to identify what molecules may be targeted for memory and in major disorders. The next step is to find how the synaptic proteome changes in neuropsychiatric diseases.” Toward New Drug Candidates In the ASSAY study, Puthanveettil and his colleagues describe their new high-throughput screening test for discovering potential drug candidates based on kinesin and axonal transport for the treatment of memory disorders. “The luminescence-based assay that we developed is highly reproducible and robust,” said Puthanveetil. Using the approach, the team screened a compound collection and identified a number of small molecules that turned on or off activity of a human kinesin. In addition to Liu and Puthanveettil, other authors of the PNAS study, “New Approach to Capture and Characterize Synaptic Proteome,” include Beena Kadakkuzha, Bruce Pascal, Caitlin Steckler,

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Komolitdin Akhmedov and Michael Chalmers of The Scripps Research Institute; and Long Yan of Max Planck Florida Institute for Neuroscience. See http://www.pnas.org/content/early/2014/10/28/1401483111.abstract This study was supported by the Whitehall Foundation and the National Institute of Mental Health of the National Institutes of Health (R21MH096258-01A1). In addition to Puthanveettil and Kadakkuzha, authors of the ASSAY study, “High-Throughput Screening for Small Molecule Modulators of Motor Protein Kinesin,” include Timothy Spicer, Peter Chase, Jeffery B. Richman and Peter Hodder (present address: Amgen, Inc.) of TSRI. See http://online.liebertpub.com/doi/abs/10.1089/adt.2014.579 This work was supported by the Alzheimer’s Drug Discovery Foundation, Margaret Q. Landenberger Research Foundation and TSRI. Scripps Florida Scientists Determine Structure of a Molecular Complex Critical for Joining Cells Together Scientists from the Florida campus of The Scripps Research Institute (TSRI) have for the first time determined the structure of a large molecular complex that plays a vital role in cell adhesion, the force that binds cells together in all animals, including humans—without it, there would be a tendency for them to simply fall apart. The new study, led by Scripps Florida Associate Professor T. Izard, was published December 8, 2014, and highlighted in an “In this Issue” article by the Journal of Cell Biology. This critical cell binding is done through specialized cell surface adhesion complexes called adherens junctions (which direct the formation of tight, Velcro-like contacts among cells), other structural proteins called F-actin (the “F” stands for filament) and focal adhesion complexes. This process is necessary for cell migration and morphogenesis, the shaping of tissues and organs that is an important part of development. In the study, the scientists produced an x-ray crystallography image of the cytoskeleton protein vinculin, an essential regulator of adherens junctions and focal adhesion, binding with a fat or lipid known as PIP2, a major component of all cell membranes. The images revealed that PIP2 binding alters vinculin structure to direct oligomerization—the linking together of a few protein or nucleic acid macromolecules—which, in turn, stabilizes focal adhesion complexes. The structural findings also revealed that vinculin’s PIP2 and actin-binding sites are distinct, suggesting that the protein may be able to bind to both molecules simultaneously. In additional experiments, the scientists determined that PIP2 binding is necessary for maintaining optimal vinculin dynamics, turnover in F-actin and for cell migration and spreading.

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The structure and functions of other lipid-and-actin-binding cytoskeleton proteins may have similar mechanisms of action for PIP2, according to the authors. In addition to Izard, authors of the study, “Lipid Binding Promotes Oligomerization and Focal Adhesion Activity of Vinculin,” were Krishna Chinthalapudi, Erumbi S. Rangarajan and Dipak N. Patil of TSRI; and Eric M. George and David T. Brown of the University of Mississippi. For more information, see http://jcb.rupress.org/content/207/5/643 The work is supported by grants from the National Institute of General Medical Sciences, the National Institutes of Health; the US Department of Defense; and by the State of Florida. Scripps Research Institute Scientists Uncover New, Fundamental Mechanism for How Resveratrol Provides Health Benefits The Ingredient Found in Red Wine Activates Ancient Stress Response Scientists at The Scripps Research Institute (TSRI) have found that resveratrol, the red-wine ingredient once touted as an elixir of youth, powerfully activates an evolutionarily ancient stress response in human cells. The finding should dispel much of the mystery and controversy about how resveratrol really works. “This stress response represents a layer of biology that has been largely overlooked, and resveratrol turns out to activate it at much lower concentrations than those used in prior studies,” said senior investigator Paul Schimmel, professor and member of the Skaggs Institute for Chemical Biology at TSRI. “With these findings we have a new, fundamental mechanism for the known beneficial effects of resveratrol,” said lead author Mathew Sajish, a senior research associate in the Schimmel laboratory. The discovery is reported in the advance online edition of Nature on December 22. Resveratrol is a compound produced in grapes, cacao beans, Japanese knotweed and some other plants in response to stresses including infection, drought and ultraviolet radiation. It has attracted widespread scientific and popular interest over the past decade, as researchers have reported that it extended lifespan and prevented diabetes in obese mice and vastly increased the stamina of ordinary mice running on wheels. More recently, though, scientists in this field have disagreed about the signaling pathways resveratrol activates to promote health, calling into question some of resveratrol’s supposed health benefits—particularly given the unrealistically high doses used in some experiments. Outsiders to the Controversy Schimmel and Sajish came to this controversy as outsiders. Schimmel’s laboratory is known for its work not on resveratrol but on an ancient family of enzymes, the tRNA synthetases. The primary and

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essential function of these enzymes is to help translate genetic material into the amino-acid building blocks that make proteins. But as Schimmel and others have shown since the late 1990s, tRNA synthetases have acquired an extensive set of added functions in mammals. Earlier Xiang-Lei Yang, a TSRI professor in the Departments of Chemical Physiology and Cell and Molecular Biology and former member of Schimmel’s laboratory, began to find hints that a tRNA synthetase called TyrRS, which links the amino acid tyrosine to the genetic material that codes for it, can move to the cell nucleus under stressful conditions—apparently taking on a protective, stress-response role. Sajish noted that resveratrol appeared to have broadly similar stress-response properties and also resembled TyrRS’s normal binding partner tyrosine. “I began to see TyrRS as a potential target of resveratrol,” he said. For the new study, Sajish and Schimmel put TyrRS and resveratrol together and showed with tests including X-ray crystallography that resveratrol does indeed mimic tyrosine, well enough to fit tightly into TyrRS’s tyrosine binding pocket. That binding to resveratrol, the team found, takes TyrRS away from its protein translation role and steers it to a function in the cell nucleus. Tracking the resveratrol-bound TyrRS in the nucleus, the researchers determined that it grabs and activates the protein, PARP-1, a major stress response and DNA-repair factor thought to have a significance influence on lifespan. The scientists confirmed the interaction in mice injected with resveratrol. TyrRS’s activation of PARP-1 led, in turn, to the activation of a host of protective genes including the tumor-suppressor gene p53 and the longevity genes FOXO3A and SIRT6. Compatible with Red Wine The first studies of resveratrol in the early 2000s had suggested that it exerts some of its positive effects on health by activating SIRT1, also thought to be a longevity gene. But SIRT1’s role in mediating resveratrol’s reported health-boosting effects has been questioned lately in terms of its particular role. The team’s experiments showed, however, that the TyrRS-PARP-1 pathway can be measurably activated by much lower doses of resveratrol—as much as 1,000 times lower—than were used in some of the more celebrated prior studies, including those focused on SIRT1. “Based on these results, it is conceivable that moderate consumption of a couple glasses of red wine (rich in resveratrol) would give a person enough resveratrol to evoke a protective effect via this pathway,” Sajish said. He also suspects that effects of resveratrol that only appear at unrealistically high doses may have confounded some prior findings. Why would resveratrol, a protein produced in plants, be so potent and specific in activating a major stress response pathway in human cells? Probably because it does much the same in plant cells, and probably again via TyrRS—a protein so fundamental to life, due to its linkage to an amino acid, that it hasn’t changed much in the hundreds of millions of years since plants and animals went their separate evolutionary ways. “We believe that TyrRS has evolved to act as a top-level switch or activator of a fundamental cell-protecting mechanism that works in virtually all forms of life,” said Sajish.

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Whatever activity resveratrol naturally has in mammals may be an example of hormesis: the mild, health-promoting activation of a natural stress response. “If resveratrol brought significant benefits to mammals, they might have evolved a symbiotic relationship with resveratrol-producing plants,” Sajish said. “We think this is just the tip of the iceberg,” said Schimmel. “We think there are a lot more amino-acid mimics out there that can have beneficial effects like this in people. And we’re working on that now.” Schimmel and his laboratory also are searching for molecules that can activate the TyrRS stress response pathway even more potently than resveratrol does. The National Cancer Institute (CA92577), the National Foundation for Cancer Research and aTyr Pharma, Inc. provided funding for the study, “A human tRNA synthetase is a potent PARP1-activating effector target for resveratrol.” For more information, see http://www.nature.com Scripps Florida Scientists Develop Novel Platform for Treatment of Breast, Pancreatic Cancer Scientists from the Florida campus of The Scripps Research Institute (TSRI) have identified a novel synthetic compound that sharply inhibits the activity of a protein that plays an important role in in the progression of breast and pancreatic cancers. In the new study, to be published in the February 2015 print edition of the journal Molecular Pharmacology, the scientists showed that the compound, known as SR1848, reduces the activity and expression of the cancer-related protein called “liver receptor homolog-1” or LRH-1. “Our study shows that SR1848 removes LRH1 from DNA, shutting down expression of LRH-1 target genes, and halts cell proliferation,” said Patrick Griffin, chair of the TSRI Department of Molecular Therapeutics and director of the Translational Research Institute at Scripps Florida. “It’s a compound that appears to be a promising chemical scaffold for fighting tumors that are non-responsive to standard therapies.” LRH1 plays a crucial role in breast cancer through its regulation of genes involved in hormone synthesis and cholesterol metabolism—also key risk factors in cardiovascular disease. LRH-1 has also been implicated as a tumor promoter in intestinal and pancreatic cancer. Overexpression of LRH-1 has been shown to promote invasiveness and metastasis, the usually lethal spread of the disease. “LRH-1 has been implicated in the proliferation and metastasis of estrogen receptor-positive breast cancers and the more difficult to treat estrogen receptor-negative breast cancers,” said Research Associate Alex Corzo, the first author of the study. “This suggests that repressing LRH-1 could be useful in treating the more aggressive triple-negative breast cancer subtype where therapies are currently so limited.”

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In fact, the study showed that levels of LHR-1 in a cell’s nucleus began to diminish four hours after treatment with SR1848, and the compound repressed specific target genes as early as two hours after administration. Griffin noted that SR1848 also appears attractive as a potential therapeutic because of its lack of impact on cells that do not express LRH1, which could mean few potential side effects. “It’s a novel mechanism that needs more study,” he said. In addition to Griffin and Corzo, other authors of the study, “Antiproliferation Activity of a Small Molecule Repressor of Liver Receptor Homolog 1s,” are Yelenis Mari, Mi Ra Chang, Tanya Khan, Dana Kuruvilla, Philippe Nuhant, Naresh Kumar, Graham M. West, Derek R. Duckett and William R. Roush of TSRI. See http://molpharm.aspetjournals.org/content/87/2/296.full This work was supported by the Intramural Research Program of the National Institutes of Health’s (NIH) National Institute of Mental Health (U54-MH074404) and National Cancer Institute (R01-CA134873). Scripps Florida Scientists Move Closer to a Personalized Treatment Solution for Intellectual Disability Scientists from the Florida campus of The Scripps Research Institute (TSRI) have produced an approach that protects animal models against a type of genetic disruption that causes intellectual disability, including serious memory impairments and altered anxiety levels. The findings, which focus on treating the effects of mutations to a gene known as Syngap1, have been published online ahead of print by the journal Biological Psychiatry. “Our hope is that these studies will eventually lead to a therapy specifically designed for patients with psychiatric disorders caused by damagingSyngap1 mutations,” said Gavin Rumbaugh, a TSRI associate professor who led the study. “Our model shows that the early developmental period is the critical time to treat this type of genetic disorder.” Damaging mutations in Syngap1 that reduce the number of functional proteins are one of the most common causes of sporadic intellectual disability and are associated with schizophrenia and autism spectrum disorder. Early estimates suggest that these non-inherited genetic mutations account for two to eight percent of these intellectual disability cases. Sporadic intellectual disability affects approximately one percent of the worldwide population, suggesting that tens of thousands of individuals with intellectual disability may carry damaging Syngap1 mutations without knowing it. In the new study, the researchers examined the effect of damaging Syngap1 mutations during development and found that the mutations disrupt a critical period of neuronal growth—a period between the first and third postnatal weeks in mouse models. “We found that a certain type of cortical neuron grows too quickly in early development, which then leads to the premature formation of certain types of neural circuits,” said Research Associate Massimilano Aceti, first author of the study.

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The researchers reasoned that this process might cause permanent errors in brain connectivity and that they might be able to head off these effects by enhancing the Syngap1 protein in the newborn mutant mice. Indeed, they found that a subset of neurons were misconnected in the adult mutant mice, suggesting that early growth of neurons can lead to life-long neural circuit connectivity problems. Then, using advanced genetic techniques to raise Syngap1 protein levels in newborn mutant mice, the researchers found this strategy completely protected the mice only when the approach was started before this critical developmental window opened. As a result of these studies, Rumbaugh and his colleagues are now developing a drug-screening program to look for drug-like compounds that could restore levels of Syngap1 protein in defective neurons. They hope that, as personalized medicine advances, such a therapy could ultimately be tailored to patients based on their genotype. In addition to Rumbaugh and Aceti, other authors of the study, “Syngap1 Haploinsufficiency Damages a Postnatal Critical Period of Pyramidal Cell Structural Maturation Linked to Cortical Circuit Assembly,” include Thomas K. Creson, Thomas Vaissiere, Camilo Rojas, Wen-Chin Huang, Ya-Xian Wang, Ronald S. Petralia, Damon T. Page and Courtney A. Miller of TSRI. For more information, see http://www.biologicalpsychiatryjournal.com/article/S0006-3223%2814%2900593-9/abstract This work was supported by the National Institutes of Health’s National Institute for Neurological Disorders and Stroke (R01NS064079), National Institute for Mental Health (R01MH096847), National Institute for Drug Abuse (R01 DA034116; R03 DA033499) and National Institute on Deafness and Other Communication Disorders/National Institutes of Health Intramural Research Program; Mrs. Nancy Lurie; and the State of Florida. Scripps Florida Scientists Establish that Drug Candidates Can Block Pathway Associated with Cell Death in Parkinson’s Disease In a pair of related studies, scientists from the Florida campus of The Scripps Research Institute (TSRI) have shown their drug candidates can target biological pathways involved in the destruction of brain cells in Parkinson's disease. The studies, published in the Journal of Medicinal Chemistry and Scientific Reports, suggest that it is possible to design highly effective and highly selective (targeted) drug candidates that can protect the function of mitochondria, which provide the cell with energy, ultimately preventing brain cell death. These drug candidates act on what are known as the JNK (pronounced “junk”) kinases—JNK1, JNK2 and JNK3—each an enzyme with a unique biological function. JNK is linked to many of the hallmark components of Parkinson's disease, such as oxidative stress and programmed cell death. “These are the first isoform selective JNK 2/3 inhibitors that can penetrate the brain and the first shown to be active in functional cell-based tests that measure mitochondrial dysfunction,” said Philip LoGrasso, a TSRI professor who led both studies. “In terms of their potential use as therapeutics,

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they’ve been optimized in every way but one—their oral bioavailability. That’s what we’re working on now.” The new studies raise the hope that such a therapy could prevent the gradual degeneration of brain cells in Parkinson's disease and halt these patients’ decline. “Some of these compounds had a level of selectivity that ranged as high as 20,000-fold against competing targets and were extremely effective against oxidative stress and mitochondrial dysfunction—both potent cell killers,” added HaJeung Park, director of Scripps Florida’s X-ray Crystallography Core Facility and the first author of the Scientific Reports study. The scientists found that within JNK3, a single amino acid—L144—was primarily responsible for the high level of JNK3 selectivity. Isoform selectivity can help to limit potential side effects of a drug. Intriguingly, some recent studies have shown that JNK3 not only plays a central role in brain cell death in Parkinson’s disease, but also in Alzheimer’s disease. LoGrasso and his colleagues also believe their JNK3 drug candidates have potential for treating ALS (Lou Gehrig’s disease). In addition to LoGrasso and Park, authors of the Scientific Reports study, “Structural Basis and Biological Consequences for JNK2/3 Isoform Selective Aminopyrazoles,” include Sarah Iqbal, Pamela Hernandez, Rudy Mora, Ke Zheng and Yangbo Feng of TSRI. See http://www.nature.com/srep/index.html The first author of the Journal of Medicinal Chemistry study, “Design and Synthesis of Highly Potent and Isoform Selective JNK32 Inhibitors: SAR Studies on Aminopyrazole Derivatives,” is Ke Zheng of TSRI. Other authors include Sarah Iqbal, Pamela Hernandez, HaJeung Park and Yangbo Feng of TSRI. See http://pubs.acs.org/doi/abs/10.1021/jm501256y Both studies were supported by the Department of Defense (W81XWH-12-1-0431 1192) and the National Institutes of Health (GM103825). Scripps Florida Scientists Discover a Key Pathway That Protects Cells Against Death by Stress When it comes to protecting cells from death brought on by the calamities of environmental stress, the human body is particularly ingenious. From cellular components that suck up misfolded proteins to a vigilant immune system, the ways we protect our cells (and ourselves) are many and mysterious. Scientists from the Florida campus of The Scripps Research Institute (TSRI) have now uncovered the workings of another cell-protection device, one that may play a major role in a number of age-related diseases, including diabetes and Parkinson’s, Alzheimer’s and Huntington’s diseases. The study, led by Srinivasa Subramaniam, a TSRI assistant professor, and Solomon H. Snyder, a neuroscience professor at Johns Hopkins University School of Medicine, was published February 5 in the journal Cell Reports.

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More or Less Acceleration The study focuses on a new pathway through which Rheb, a regulator that many believe is active in the brain’s ability to change in response to learning, actually plays two roles, rather than one—stimulating and inhibiting protein synthesis. The interplay between the two roles may be the key that enables cells to alter protein synthesis and protect the cell in response to varying environmental stresses. “We found Rheb acts like the gas pedal in a car,” Subramaniam said. “It can either increase translation or decrease it. And because translation is a fundamental process that is affected in a lot of diseases, we now think that Rheb may act like a switch in some disease states—helping to turn them off and on.” Rheb is known to bind and activate mTOR, a developmentally important gene that integrates signals from multiple pathways and regulates critical cell functions such as protein synthesis. Besides its role as an activator of mTOR, which plays a major role in conditions from diabetes to neurodegenerative disease, the mTOR-independent role of Rheb is less known. The new study defines crucial mTOR-independent effects of Rheb. Results showed that, when stressed, Rheb instead inhibits protein synthesis by amplifying the phosphorylation (adding a phosphate group to a protein to alter its function) of another protein known eIF2a. As a result, cell resources can be conserved rather than squandered when the environment is challenging. “We don’t really understand the full role of the Rheb-mTOR pathway, but we have uncovered a new fundamental process of Rheb that is independent of mTOR and very intriguing,” said Neelam Shahani, a member of Subramaniam’s lab who was co-first author of the study with Richa Tyagi of Johns Hopkins University School of Medicine. “Rheb can inhibit protein synthesis, and we know that protein misfolding via environmental stress factors is present in a lot of diseases.” Subramaniam noted that, intriguingly, an earlier study had suggested the Rheb pathway had been implicated in Alzheimer’s disease. “We also want to look at Rheb’s role in other neurodegenerative diseases,” he said. In addition to Subramaniam, Snyder, Shahani and Tyagi, authors of the study, “Rheb Inhibits Protein Synthesis by Activating the PERK-eIF2α Signaling Cascade,” include Max Ferretti, William Pryor, Supriya Swarnkar and Katrin Karbstein of TSRI; Lindsay Gorgen of Florida Atlantic University; as well as Paul F. Worley and Po Yu Chen of Johns Hopkins University School of Medicine. This work was supported by the State of Florida, the O'Keeffe Neuroscience Scholar Award and the United States Public Health Service (DA000266).

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Scripps Florida Scientists' 'Mad Cow' Discovery Points to Possible Neuron Killing Mechanism Behind Alzheimer’s and Parkinson’s Diseases $1.4 Million Grant Will Enable Team to Follow Up with Search for Drug Candidates Scientists from the Florida campus of The Scripps Research Institute (TSRI) have for the first time discovered a killing mechanism that could underpin a range of the most intractable neurodegenerative diseases such as Alzheimer’s, Parkinson’s and ALS. The new study, published recently in the journal Brain, revealed the mechanism of toxicity of a misfolded form of the protein that underlies prion diseases, such as bovine spongiform encephalopathy (“mad cow disease”) and its human equivalent, Creutzfeldt-Jakob disease. “Our study reveals a novel mechanism of neuronal death involved in a neurodegenerative protein-misfolding disease,” said Corinne Lasmézas, a TSRI professor who led the study. “Importantly, the death of these cells is preventable. In our study, ailing neurons in culture and in an animal model were completely rescued by treatment, despite the continued presence of the toxic misfolded protein. This work suggests treatment strategies for prion diseases—and possibly other protein misfolding diseases such as Alzheimer’s.” Failure and Rescue of Brain Cells In the new study, the scientists used a misfolded form of the prion disease protein, called TPrP, a model they had previously developed, to study misfolded protein-induced neurodegeneration in the laboratory. Misfolded proteins are the common cause of the group of diseases comprising prion, Alzheimer’s, Parkinson’s diseases, ALS and other conditions. Using biochemical techniques, the researchers demonstrated that TPrP induces neuronal death by profoundly depleting NAD+ (nicotinamide adenine dinucleotide)—a metabolite well known as a coenzyme that is common to all cells and necessary for energy production and cellular homeostasis. Restoring NAD+ proved to be the critical factor for the rescue of neurons subjected to TPrP injury. Even when added three days after TPrP exposure, an infusion of NAD+ reversed within only a few hours the fate of neurons that had been doomed to destruction. “Our study shows for the first time that a failure of NAD+ metabolism is the cause of neuronal loss following exposure to a misfolded protein,” Lasmézas said. The loss of NAD+ also triggers autophagy—a way cells rid themselves of damaged material such as misfolded proteins—and apoptosis, or programmed cell death, the last resort of the cell when everything starts to go wrong. However, the researchers demonstrated these mechanisms do not initiate the neuronal demise. “We show that apoptosis or programmed cell death and autophagy are not primary players in the death cascade,” said Staff Scientist Minghai Zhou, the first author of the study. “Modulation of neither of these processes significantly alters the death of TPrP-exposed neurons. This is all caused by NAD+ disappearing—the cell cannot survive without it.”

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Lasmézas noted the loss of NAD+ is suggestive of some other neurodegenerative diseases, such as Parkinson’s where NAD+ depletion could play a role in mitochondrial failure. New Grant to Support Further Research A recent $1.4-million grant from the National Institute of Neurological Disorders and Stroke (NINDS) will support further work to look for drug candidates based on the new findings. Lasmézas and Louis Scampavia, a TSRI associate professor of molecular therapeutics, will be co-principal investigators for the new three-year study, whose team will also include Tom Bannister, a TSRI associate scientific director at Scripps Florida’s Translational Research Institute. The scientists have developed several primary tests for compounds that could restore NAD+ and plan to begin those tests at Scripps Florida’s High Throughput Screening facility. Since it was established in 2005, the Scripps Florida High Throughput Screening facility has screened more than 200 targets in more than 235 industrial and academic collaborations—several of these collaborations have produced successful clinical trial candidates. The drug discovery facility is currently capable of routinely screens one quarter of a million compounds in a single day. In addition to Zhou and Lasmézas, other authors of the study, “Neuronal Death Induced by Misfolded Prion Protein Is Due To NAD+ Depletion and Can Be Relieved In Vitro And In Vivo by NAD+ Replenishment,” include Gregory Ottenberg, Gian Franco Sferrazza, Christopher Hubbs, Mohammad Fallahi, Gavin Rumbaugh and Alicia F. Brantley of TSRI. The work was supported by TSRI and by the National Institutes of Health (RNS081519). The number of the new NINDS grant is 1R01NS085223. Microbes Prevent Malnutrition in Fruit Flies—and Maybe Humans, Too Microbes, small and ancient life forms, play a key role in maintaining life on Earth. As has often been pointed out, without microbes, we’d die—without us, most microbes would get along just fine. Now, a study by scientists from the Florida campus of The Scripps Research Institute (TSRI) sheds significant new light on a surprising and critical role that microbes may play in nutritional disorders such as protein malnutrition. Using fruit flies—Drosophila melanogaster—as a simple and easily studied stand-in for humans, these new findings advance our understanding of the fundamental mechanisms underlying microbial contributions to metabolism and may point to long-term strategies to treat and prevent malnutrition in general.

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In the study, published February 12 in the journal Cell Reports, a team led by TSRI biologist William Ja showed that Issatchenkia orientalis, a fungal microbe isolated from field-caught fruit flies, promotes nutritional harvest that rescues the health and longevity of undernourished flies. Surprising Protein Harvest Using a range of radioisotope-labeled dietary components such as amino acids (the components of proteins and the basic building blocks of the body) and sucrose (sugar) to measure the transfer of nutrients from food to microbe to fly, the study shows that the microbes first harvest amino acids directly from the fly’s food sources and then transfer that protein to the fly—by being eaten. “Flies in the wild carry microbes to every surface they touch,” said Research Associate Ryuichi Yamada, who spearheaded the study in the Ja lab. “As flies land on low-protein fruit, they deposit microbes, which take up and concentrate the available amino acids. By eating the microbes, flies gain a much needed source of dietary protein.” In flies that are fed nutrient-poor diets, this chain of events restores body mass and protein levels, essentially returning them to the pre-malnutrition profile of well-fed flies. “Ryuichi and colleagues did a lot of painstaking work to carefully show that the simplest explanation for what was happening was correct,” Ja said. “The direct influence of microbes on fly nutrition is often overlooked and may be relevant in numerous studies of host-microbe interactions.” Natural Symbiosis This relationship appears to be particularly beneficial for flies. Devouring the protein-plumped microbes extends fly lifespan during periods when nutrients are scarce. “In fact, the I. orientalis microbe is commonly found in field-trapped fruit flies,” said Yamada. “That suggests a natural symbiosis.” Ja believes the study also offers a larger lesson on the partnership that can occur between microorganisms and their hosts, in addition to providing information on nutrient harvesting and the potential of Drosophila as a platform for studies of host-microbe relationships. “While everyone keeps looking for that single magic microbial metabolite or species, what has been increasingly ignored is the bulk effect that microbes have on primary metabolism,” he said. “Our study suggests that diverse [microbial] species could each benefit their hosts and that their quantity, rather than quality, may be of fundamental importance.” In addition to Ja and Yamada, other authors of the study, “Microbes Promote Amino Acid Harvest to Rescue Undernutrition in Drosophila,” are Sonali A. Deshpande, Kimberley D. Bruce and Elizabeth M. Mak of TSRI. For more information, see http://www.cell.com/cell-reports/home The work was supported by the National Institutes of Health (grants R00AG030493 and R21DK092735), The Ellison Medical Foundation and the Glenn Foundation for Medical Research.

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Scripps Florida Scientists Announce Anti-HIV Agent So Powerful It Can Work in a Vaccine In a remarkable new advance against the virus that causes AIDS, scientists from The Scripps Research Institute (TSRI) have announced the creation of a novel drug candidate that is so potent and universally effective, it might work as part of an unconventional vaccine. The research, which involved scientists from more than a dozen research institutions, was published February 18 online ahead of print by the prestigious journal Nature. The study shows that the new drug candidate blocks every strain of HIV-1, HIV-2 and SIV (simian immunodeficiency virus) that has been isolated from humans or rhesus macaques, including the hardest-to-stop variants. It also protects against much-higher doses of virus than occur in most human transmission and does so for at least eight months after injection. “Our compound is the broadest and most potent entry inhibitor described so far,” said Michael Farzan, a professor on TSRI's Florida campus who led the effort. “Unlike antibodies, which fail to neutralize a large fraction of HIV-1 strains, our protein has been effective against all strains tested, raising the possibility it could offer an effective HIV vaccine alternative.” Blocking a Second Site When HIV infects a cell, it targets the CD4 lymphocyte, an integral part of the body’s immune system. HIV fuses with the cell and inserts its own genetic material—in this case, single-stranded RNA—and transforms the host cell into a HIV manufacturing site. The new study builds on previous discoveries by the Farzan laboratory, which show that a co-receptor called CCR5 contains unusual modifications in its critical HIV-binding region, and that proteins based on this region can be used to prevent infection. With this knowledge, Farzan and his team developed the new drug candidate so that it binds to two sites on the surface of the virus simultaneously, preventing entry of HIV into the host cell. “When antibodies try to mimic the receptor, they touch a lot of other parts of the viral envelope that HIV can change with ease,” said TSRI Research Associate Matthew Gardner, the first author of the study with Lisa M. Kattenhorn of Harvard Medical School. “We’ve developed a direct mimic of the receptors without providing many avenues that the virus can use to escape, so we catch every virus thus far.” The team also leveraged preexisting technology in designing a delivery vehicle—an engineered adeno-associated virus, a small, relatively innocuous virus that causes no disease. Once injected into muscle tissue, like HIV itself, the vehicle turns those cells into “factories” that could produce enough of the new protective protein to last for years, perhaps decades, Farzan said. Data from the new study showed the drug candidate binds to the envelope of HIV-1 more potently than the best broadly neutralizing antibodies against the virus. Also, when macaque models were inoculated with the drug candidate, they were protected from multiple challenges by SIV.

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“This is the culmination of more than a decade’s worth of work on the biochemistry of how HIV enters cells,” Farzan said. “When we did our original work on CCR5, people thought it was interesting, but no one saw the therapeutic potential. That potential is starting to be realized.” In addition to Farzan, Gardner and Kattenhorn, authors of the study, “AAV-expressed eCD4-Ig provides durable protection from multiple SHIV challenges,” include Hema R. Kondur, Tatyana Dorfman, Charles C. Bailey, Christoph H. Fellinger, Vinita R. Josh and Brian D. Quinlan of TSRI; Dennis R. Burton of the Department of Immunology and Microbial Science, the International AIDS Vaccine Initiative’s (IAVI) Neutralizing Antibody Center, and the Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery (CHAVI-ID) at TSRI, and the Ragon Institute; Pascal Poignard of the Department of the Immunology and Microbial Science, IAVI Neutralizing Antibody Center, and CHAVI-ID at TSRI; Jessica J. Chiang and Annie Y. Yao of Harvard Medical School; Michael D. Alpert of Harvard Medical School and Immunathon Inc.; Ronald C. Desrosiers of Harvard Medical School and the University of Miami Miller School of Medicine; Kevin G. Haworth and Paula M. Cannon of the University of Southern California; Julie M. Decker and Beatrice H. Hahn of the University of Pennsylvania; Sebastian P. Fuchs and Jose M. Martinez-Navio of the University of Miami Miller School of Medicine; Hugo Mouquet of The Rockefeller University and Institut Pasteur; Michel C. Nussenzweig of The Rockefeller University and Howard Hughes Medical Center; Jason Gorman, Baoshan Zhang and Peter D. Kwong of the National Institutes of Health; Michael Piatak Jr. and Jeffrey D. Lifson of the Frederick National Laboratory for Cancer Research; Guangping Gao of the University of Massachusetts Medical School; David T. Evans of the University of Wisconsin; and Michael S. Seaman of Beth Israel Deaconess Medical Center. The work was supported by the National Institutes of Health (grants R01 AI091476, R01 AI080324, P01 AI100263, RR000168 and R01AI058715). New Study Shows Decreased Aggressive Behavior Toward Strangers in Autism Spectrum Disorder Model While aggression toward caregivers and peers is a challenge faced by many individuals and families dealing with autism, there has been much speculation in the media over the possibility of generally heightened aggression in those diagnosed with autism spectrum disorder. A new study by scientists from the Florida campus of The Scripps Research Institute (TSRI) found no evidence of increased aggressive behavior toward strangers in an animal model of the condition. In fact, the study, published recently online ahead of print in the journal Genes, Brain and Behavior, found these animals showed decreased aggressive behavior toward strangers and, instead, engage in more repetitive behavior than normal mice. “These mice show traits relevant to autism, such as an overgrown brain and reduced social interaction,” said Damon Page, a TSRI biologist who conducted the study with Research Associate Amy Clipperton Allen. “What we don’t see in this model is a general increase in aggressive behavior.”

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Autism spectrum disorder is a highly inheritable condition characterized by impaired social behavior and communication skills and a tendency towards repetitive patterns of behavior. A 2010 survey of eight-year-olds in 11 communities across the United States by the Centers for Disease Control and Prevention (CDC) found a rate of autism spectrum disorder of approximately one in 68 children. Boys, it found, are at four- to five-times greater risk than girls. Page and his colleagues, who use animal models to understand how autism risk factors impact the developing brain and to identify potential treatments for the condition, have found that animals with mutations in the autism risk gene phosphatase and tensin homolog (Pten) mimic aspects of autism, including increased brain size, social deficits and increased repetitive behavior. The new study used the model to examine aggressive behavior directed at unfamiliar mice. Typically, when an unfamiliar mouse is put into the “home” cage of another mouse, the resident will attack the intruder, reflecting the natural tendency of mice to defend their territory. That was not the case in the study. When a stranger mouse was placed in the home of a Pten-mutant mouse, instead of attacking or investigating the intruder, these mutant animals engaged in repetitive behavior—in this case, digging. “This is a striking result,” Page said. “The mutant mice appear to avoid aggressive encounters with the intruder and instead engage in repetitive behavior. An analogy might be a stranger entering a house, and the resident rearranging the books on the bookshelf instead of confronting the intruder.” Page plans further studies examining the neurobiological relationship between social deficits and repetitive behavior, two of the primary symptoms of autism. The study, “Decreased Aggression and Increased Repetitive Behavior in Pten Haploinsufficient Mice,” was supported by Ms. Nancy Lurie Marks, The American Honda and Children’s Healthcare Charity Inc. and the State of Florida. For more information on the study, see http://onlinelibrary.wiley.com/doi/10.1111/gbb.12192/abstract FAU, Scripps Florida, Max Planck Announce Plans for Groundbreaking Research and Education Collaboration One of Florida’s leading public research universities and two of the world’s premier research institutions will create one-of-a-kind education programs that will attract the best and brightest students to Palm Beach County, and transform Florida Atlantic University’s John D. MacArthur Campus in Jupiter into a hub of scientific inquiry, innovation and economic development. FAU, and the globally acclaimed Max Planck Florida Institute and The Scripps Research Institute, will build on existing relationships to further scientific discovery and education through shared resources and facilities.

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The three institutions will provide undergraduate and graduate students the unprecedented opportunity to enroll in unique degree programs in collaboration with Max Planck and Scripps Florida at the MacArthur Campus. The initiative will allow students to work and study alongside some of the world’s leading scientific researchers as part of their degree programs, while undergraduate research projects will be mentored by these same scientists. The Institutes will collaborate to develop premier STEM programs — Science, Technology, Engineering, Math — and combine FAU Jupiter’s existing strengths in STEM areas, with support from the arts, to create a leading STEAM initiative. FAU President John Kelly said the alliance will help cure diseases, develop drugs, educate students and generate jobs. FAU’s economic impact on Florida’s economy during 2010-2011, the most recently available data, was $6.3 billion. This initiative creates unique opportunities for FAU’s colleges of science, medicine, and engineering and computer science to greatly increase that number, Kelly said. “This initiative comes from the core of economic development,” Kelly said. “FAU, Max Planck and Scripps will solve real-world problems and take strides to improve human health. “We will create the knowledge economy of the future,” he said. “Moreover, we will provide students unique scientific research programs that will be the envy of the world.” A shared facilities environment will provide students access to state-of-the-art scientific equipment. Max Planck and Scripps Florida researchers will have access to FAU faculty, teaching space, and research equipment. James Paulson, acting president and CEO of The Scripps Research Institute, said the Scripps mission is to build a world-class biomedical research presence in Florida for the benefit of human health and to train the next generation of scientists. “We believe this new agreement strengthens our existing collaboration with FAU and the Max Planck Institute and enables us to work more closely with our local partners to achieve these critical goals,” Paulson said. David Fitzpatrick, CEO and scientific director at Max Planck, said, importantly, the collaboration will increase research funding in areas of common interest. The Max Planck Florida Institute’s research focus is neuroscience, specifically, gaining insights into brain circuitry. The institute utilizes some of the world’s most advanced technologies in brain research. “Combining our resources makes this collaboration a potent force in the scientific and healthcare fields,” Fitzpatrick said. “The advances we can take in many important research areas will be significant. “Together, FAU, Max Planck and Scripps will train the scientific leaders of tomorrow,” he said.

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Scripps Florida Scientists Find a Defect Responsible for Memory Impairment in Aging Everyone worries about losing their memory as they grow older—memory loss remains one of the most common complaints of the elderly. But the molecular reasons behind the processes remain unclear, particularly those associated with advancing age. Now, scientists from the Florida campus of The Scripps Research Institute (TSRI) have discovered a mechanism that causes long-term memory loss due to age in Drosophila, the common fruit fly, a widely recognized substitute for human memory studies. The new study, published recently in The Journal of Neuroscience, describes in detail the loss of connectivity between two sets of neurons that prevents the formation of long-term memory. “We show how long-term memory is impaired with age in Drosophila,” said Ron Davis, a TSRI professor and chair of the Department of Neuroscience who led the study. “This isn’t due to any functional defects, but to connectivity problems between neurons.” The most widely studied form of memory in fruit flies is memory of smell. When an odor is paired with a mild electric shock, the flies develop short-term memories that persist for around a half-hour, intermediate-term memory that lasts a few hours and long-term memory that persists for days. Using real-time cellular imaging to monitor the changes in aged flies’ neuron activity before and after learning, Davis and his colleague Ayako Tonoki found structural connectivity defects between a set of neurons known as dorsal paired medial neurons and mushroom body neurons; these defects prevented long-term memories from forming. Long-term memories require new synapses and new proteins to be formed—as compared to short-term memory, which is built from existing proteins. “Now that we know long-term memory loss is a connection problem,” said Davis, “to improve memory we’re going to have to think of ways of rebuilding those connections.” The study, “Aging Impairs Protein-Synthesis-Dependent Long-Term Memory in Drosophila,” was supported by the National Institutes of Health’s National Institute of Neurological Disorders and Stroke (grant R37 NS19904) and by the Japan Society for the Promotion of Science (KAKENHI Grants 25115703, 26115505 and 26830003). For more information on the study, see http://www.jneurosci.org/content/35/3/1173.short Scripps Research, Mayo Clinic Scientists Find New Class of Drugs that Dramatically Increases Healthy Lifespan A research team from The Scripps Research Institute (TSRI), Mayo Clinic and other institutions has identified a new class of drugs that in animal models dramatically slows the aging process—alleviating symptoms of frailty, improving cardiac function and extending a healthy lifespan.

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The new research was published March 9 online ahead of print by the journal Aging Cell. The scientists coined the term “senolytics” for the new class of drugs. “We view this study as a big, first step toward developing treatments that can be given safely to patients to extend healthspan or to treat age-related diseases and disorders,” said TSRI Professor Paul Robbins, PhD, who with Associate Professor Laura Niedernhofer, MD, PhD, led the research efforts for the paper at Scripps Florida. “When senolytic agents, like the combination we identified, are used clinically, the results could be transformative.” “The prototypes of these senolytic agents have more than proven their ability to alleviate multiple characteristics associated with aging,” said Mayo Clinic Professor James Kirkland, MD, PhD, senior author of the new study. “It may eventually become feasible to delay, prevent, alleviate or even reverse multiple chronic diseases and disabilities as a group, instead of just one at a time.” Finding the Target Senescent cells—cells that have stopped dividing—accumulate with age and accelerate the aging process. Since the “healthspan” (time free of disease) in mice is enhanced by killing off these cells, the scientists reasoned that finding treatments that accomplish this in humans could have tremendous potential. The scientists were faced with the question, though, of how to identify and target senescent cells without damaging other cells. The team suspected that senescent cells’ resistance to death by stress and damage could provide a clue. Indeed, using transcript analysis, the researchers found that, like cancer cells, senescent cells have increased expression of “pro-survival networks” that help them resist apoptosis or programmed cell death. This finding provided key criteria to search for potential drug candidates. Using these criteria, the team homed in on two available compounds—the cancer drug dasatinib (sold under the trade name Sprycel®) and quercetin, a natural compound sold as a supplement that acts as an antihistamine and anti-inflammatory. Further testing in cell culture showed these compounds do indeed selectively induce death of senescent cells. The two compounds had different strong points. Dasatinib eliminated senescent human fat cell progenitors, while quercetin was more effective against senescent human endothelial cells and mouse bone marrow stem cells. A combination of the two was most effective overall. Remarkable Results Next, the team looked at how these drugs affected health and aging in mice. “In animal models, the compounds improved cardiovascular function and exercise endurance, reduced osteoporosis and frailty, and extended healthspan,” said Niedernhofer, whose animal models

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of accelerated aging were used extensively in the study. “Remarkably, in some cases, these drugs did so with only a single course of treatment.” In old mice, cardiovascular function was improved within five days of a single dose of the drugs. A single dose of a combination of the drugs led to improved exercise capacity in animals weakened by radiation therapy used for cancer. The effect lasted for at least seven months following treatment with the drugs. Periodic drug administration of mice with accelerated aging extended the healthspan in the animals, delaying age-related symptoms, spine degeneration and osteoporosis. The authors caution that more testing is needed before use in humans. They also note both drugs in the study have possible side effects, at least with long-term treatment. The researchers, however, remain upbeat about their findings’ potential. “Senescence is involved in a number of diseases and pathologies so there could be any number of applications for these and similar compounds,” Robbins said. “Also, we anticipate that treatment with senolytic drugs to clear damaged cells would be infrequent, reducing the chance of side effects.” The co-first authors of the study, “Achilles’ Heel of Senescent Cells: From Transcriptome to Senolytic Drugs,” are Yi Zhu and Tamara Tchkonia of the Mayo Clinic. In addition to Robbins, Niedernhofer and Kirkland, other authors include Sara J. McGowan, Heike Fuhrmann-Stroissnigg, Aditi Gurkar, Jing Zhao, Debora Colangelo, Akaitz Dorronsoro, Yuan Yuan Ling, Amira Barghouthy, Diana Navarro and Tokio Sano of TSRI; Yuji Ikeno and Gene Borden of The University of Texas Health Science Center; Adam Gower and Marc Lenburg of Boston University; Yi Zhu (co-first author), Tamara Tchkonia (co-first author), Tamar Pirtskhalava, Husheng Ding, Nino Giorgadze, Allyson Palmer, Steven O'Hara, Nicholas LaRusso, Carolyn Roos, Jordan Miller, Carolyn Roos, Grace Verzosa, Nathan LeBrasseur, Joshua Farr, Sundeep Khosla and Michael Stout of Mayo Clinic; and Jonathan Wren of Oklahoma Medical Research Foundation. See http://onlinelibrary.wiley.com/doi/10.1111/acel.12344/abstract The work was supported by the National Institutes of Health (grants AG013925, AG041122, AG031736, AG044396, DK050456, HL111121 and AG043376), the Glenn Foundation and the Clinical & Translational Science Awards (grant UL1-TR000157). Scripps Florida Scientists Confirm Key Targets of New Anti-Cancer Drug Candidates Ribosomes, ancient molecular machines that produce proteins in cells, are required for cell growth in all organisms, accomplishing strikingly complex tasks with apparent ease. But defects in the assembly process and its regulation can lead to serious biological problems, including cancer. Now, in a study published in the March 16 issue of The Journal of Cell Biology, scientists from the Florida campus of The Scripps Research Institute (TSRI) have confirmed the ribosome assembly process as a potentially fertile new target for anti-cancer drugs by detailing the essential function of a key component in the assembly process.

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“This study confirms that ribosome assembly is a good therapeutic target in cancer,” said Katrin Karbstein, a TSRI associate professor who led the study. “Whether or not we have pinpointed the best molecule remains to be shown, but this is a vindication of our basic research. There should be effort devoted to exploring this pathway.” Understanding ribosome assembly—which involves about 200 essential proteins known as "assembly factors" in addition to the four RNA molecules and 78 ribosomal proteins that are part of the mature ribosome—has become a fruitful area of research in recent years because of the importance of ribosome assembly for cell growth. The new study highlights the molecules Casein kinase 1δ (CK1δ) and CK1ε, which are essential for human ribosome assembly. The expression of CK1δ is elevated in several tumor types, as well as Alzheimer’s and Parkinson’s disease—and CK1δ inhibitors have shown promise in some pre-clinical animal studies. In the new study, Karbstein and her group—working closely with three labs across the state of Florida, including the laboratory of William Roush at Scripps Florida—used Hrr25, the yeast equivalent of Casein kinase 1δ (CK1δ) and CK1ε, as a research model. In biochemical experiments, the team showed that Hrr25 is necessary for ribosome assembly and that the molecule normally adds a phosphate group to an assembly factor called “Ltv1,” allowing it to separate from other subunits and mature. If Hrr25 is inactivated or a mutation blocks the release of Ltv1, the assembly process is doomed. “Inhibiting Hrr25 and the subsequent release of Ltv1 blocks the formation of other subunits that are required for maturation—and the subsequent production of proteins,” said Homa Ghalei, the first author of the study and a member of the Karbstein lab. In additional experiments on human breast cancer cells, the researchers showed that CK1δ/CK1ε inhibitors no longer induce programmed cell death (“apoptosis”) and prevent cancer cells from growing when Ltv1 is removed. “This clearly establishes that the anti-proliferative potency of these inhibitors is in large part due to blocking ribosome assembly,” Karbstein said. In addition to Karbstein and Ghalei, other authors of the study, “Hrr25/CK1d-Directed Release of Ltv1 From Pre-40S Ribosomes Is Necessary For Ribosome Assembly And Cell Growth” (10.1083/jcb.201409056), are Joanne R. Doherty, Yoshihiko Noguchi and William R. Roush of TSRI; Franz X. Schaub and John L. Cleveland of The Moffitt Cancer and Research Institute; and M. Elizabeth Stroupe of Florida State University. See http://jcb.rupress.org/content/208/6/745.abstract The work was supported by the National Institutes of Health (grants R01-GM086451, CA154739, U54MH074404 and P30-CA076292), the National Science Foundation (grant 1149763), the ThinkPink Kids Foundation, the PGA National Women’s Cancer Awareness Days and the Swiss National Foundation (P300P3-147907).

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New Compound Prevents Type 1 Diabetes in Animal Models—Before It Begins Scientists from the Florida campus of The Scripps Research Institute (TSRI) have successfully tested a potent synthetic compound that prevents type 1 diabetes in animal models of the disease. “The animals in our study never developed high blood sugar indicative of diabetes, and beta cell damage was significantly reduced compared to animals that hadn’t been treated with our compound,” said Laura Solt, Ph.D., a TSRI biologist who was the lead author of the study. Type 1 diabetes is a consequence of the autoimmune destruction of insulin-producing beta cells in the pancreas. While standard treatment for the disease aims to replace lost insulin, the new study focuses instead on the possibility of preventing the initial devastation caused by the immune system—stopping the disease before it even gets started. In the study, published in the March 2015 issue of the journal Endocrinology, the scientists tested an experimental compound known as SR1001 in non-obese diabetic animal models. The compound targets a pair of “nuclear receptors” (RORα and RORg) that play critical roles in the development of a specific population (Th17) of immune cells associated with the disease. “Because Th17 cells have been linked to a number of autoimmune diseases, including multiple sclerosis, we thought our compound might inhibit Th17 cells in type 1 diabetes and possibly interfere with disease progression,” said Solt. “We were right.” The researchers found SR1001 eliminated the incidence of diabetes and minimized insulitis, which is the inflammation associated with, and destroyer of, insulin-producing cells, in the treated animals. The compound suppressed the immune response, including the production of Th17 cells, while maintaining normal insulin levels; it also increased the frequency of the expression of Foxp3 in T cells, which controls the development and function of a type of immune cell known as T regulatory cells. Solt notes that the study strongly suggests that Th17 cells have a pathological role in the development of type 1 diabetes and use of ROR-specific synthetic compounds targeting this cell type may have potential as a preventative therapy for type 1 diabetes. “It certainly opens the door for other areas to be looked at,” she said. Other authors of the study, “ROR Inverse Agonist Suppresses Insulitis and Prevents Hyperglycemia in a Mouse Model of Type 1 Diabetes,” include Subhashis Banerjee, Sean Campbell and Theodore M. Kamenecka of The Scripps Research Institute, and Thomas Burris of Saint Louis University School of Medicine. For more information on the study, see http://press.endocrine.org/doi/pdf/10.1210/en.2014-1677 This work was supported by National Institutes of Health (grants DK080201, MH092769 and DK089984) and a National Research Service Award (DK088499).

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Scripps Florida Scientists Reveal Unique Mechanism of Natural Product with Powerful Antimicrobial Action Scientists from the Florida campus of The Scripps Research Institute (TSRI) have uncovered the unique mechanism of a powerful natural product with wide-ranging antifungal, antibacterial, anti-malaria and anti-cancer effects. The new study, published online ahead of print by the journal Nature Communications, sheds light on the natural small molecule known as borrelidin. “Our study may help the rational design of compounds similar to borrelidin with a range of useful applications, particularly in cancer,” said Min Guo, a TSRI associate professor who led the study. Powerful Medicines Guo and his colleagues were interested in borrelidin because it inhibits a specific type of enzyme known as threonyl-tRNA synthetase (ThrRS), ultimately impeding protein synthesis. Compounds similar to borrelidin have been used as treatments for microbial infections. For example, the natural product mupirocin is approved as a topical treatment for bacterial skin infections and febrifugine (the active component of the Chinese herb Chang Shan (Dichroa febrifuga Lour)) has been used for treating malaria-induced fever for nearly 2,000 years. Previous studies from the collaborator Professor Christopher S. Francklyn of the University of Vermont College of Medicine and others have shown that borrelidin impedes angiogenesis, the growth of new blood vessels critical for the spread of malignant tumors, as well as increasing apoptosis in certain types of leukemia. “It is probably the most potent tRNA synthetase inhibitor on Earth,”said Research Associate Pengfei Fang, co-first author of the study and member of the Guo lab at Scripps Florida. “It is also the earliest known tRNA synthetase inhibitor, discovered in 1966—just a few years after people learned the existence of tRNA synthetase and genetic code.” Research Associate Xue Yu, also co-first author of the study and a member of the Guo lab, emphasized, “While little is known about how borrelidin works, the fairly widespread use of these types of inhibitors highlights their tremendous potential in a number of medical applications.” Winning at Musical Chairs In the new study, the scientists set out to conduct a detailed structural and functional analysis of the binding of borrelidin to both human and bacterial (E. coli) ThrRS in the hope of identifying its unique mechanism. The researchers succeeded, and the new study shows for the first time that borrelidin occupies four distinct subsites on both the bacterial and human tRNA synthetase, including all three subsites for its normal binding substrates and an extra one that is created when the compound binds. In this way,

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borrelidin crowds out all natural partners that would otherwise bind those sites and fuel the process of protein synthesis. In that sense, borrelidin more or less wins the game of molecular musical chairs by taking over everyone’s seat well before the music starts, even including the aisles. Because each of the subsites is essential for its activity, the fact that borrelidin occupies four subsites within ThrRS, an apparent inhibitory overkill, was a quite surprise, and indeed accounts for its potency as validated by further experiments done in both in vitro and in cells. “This has never been seen in any other tRNA synthetase inhibitors, including the ones sold as medicines,” said Guo. “This finding establishes a new inhibitor class and highlights the striking design of this natural compound that inhibits tRNA synthetases in two of the three kingdoms of life.” In addition to Guo, Fang and Yu, other authors of the study, “Structural Basis for Full-Spectrum Inhibition of Translational Functions on a tRNA Synthetase,” are Kaige Chen and Xin Chen of TSRI; Seung Jae Jeong and Sunghoon Kim of Seoul National University, Korea; and Adam Mirando and Christopher S. Francklyn of the University of Vermont College of Medicine. The work was supported by the National Institutes of Health (grants NIEHS T32 ES007122-23, GM54899, GM100136 and GM106134), the Korean Global Frontier Project (NRF-M1AXA002-2010-0029785), and the PGA Women’s Cancer Awareness Foundation. Scripps Florida Scientists Uncover How Molecule Protects Brain Cells in Parkinson’s Disease Model Scientists from the Florida campus of The Scripps Research Institute (TSRI) have found how a widely known but little-studied enzyme protects brain cells in models of Parkinson’s disease. These findings could provide valuable insight into the development of drug candidates that could protect brain cells in Parkinson’s and other neurodegenerative diseases. The study, published recently online ahead of print by the journal Molecular and Cellular Biology, focuses on the enzyme known as serum glucocorticoid kinase 1 (SGK1). “The overexpression of SGK1 provides neuron protection in both cell culture and in animal models,” said Philip LoGrasso, a TSRI professor who led the study. “It decreases reactive oxygen species generation and alleviates mitochondrial dysfunction.” Using a neurotoxin animal model of neurodegeneration, the study showed that SGK1 protects brain cells by blocking several pathways involved in neurodegeneration, deactivating other molecules known as JNK, GSK3β and MKK4. Increasing SGK1 offers a potential therapeutic strategy because, as the study makes clear, there isn’t enough naturally occurring SGK1 to do the job.

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“Even though the levels of naturally occurring SGK1 increases in the cell under stress, it was not enough to promote cell survival in our neurodegeneration model,” said Sarah Iqbal, the first author of the study and a member of the LoGrasso lab. “On the other hand, cell survival mechanisms tend to dominate when more SGK1 is added to the neurons.” The LoGrasso lab plans to continue to explore SGK1 as a therapeutic possibility for Parkinson’s disease. In addition to LoGrasso and Iqbal, other authors of the study, “Serum-Glucocorticoid-Inducible Kinase 1 Confers Protection in Cell-Based and in In Vivo Neurotoxin Models Via the C-Jun N-Terminal Kinase Signaling Pathway,” include Shannon Howard of TSRI. For more information on the study, see http://mcb.asm.org/content/early/2015/03/27/MCB.01510-14.full.pdf The work was supported by the Department of Defense (grant W81XWH-12-1-0431), the National Institutes of Health (grants U01-NS057153 and GM103825), the Michael J Fox Foundation/23&Me, the Saul and Theresa Esman Foundation and a gift from the McCubbin Family. Scripps Florida Scientists Uncover Surprising New Details of Potential Alzheimer’s Treatment Taking a new approach, scientists from the Florida campus of The Scripps Research Institute (TSRI) have uncovered some surprising details of a group of compounds that have shown significant potential in stimulating the growth of brain cells and memory restoration in animal models that mimic Alzheimer’s disease. The new study points to promising new directions using a known therapeutic strategy for Alzheimer’s disease—a disorder that will affect nearly 14 million Americans by 2050, according to the Alzheimer’s Association. The study, which was led by TSRI Associate Professors Courtney Miller and Gavin Rumbaugh, appears online ahead of print in the journal Neuropsychopharmacology. This new study builds on previous findings from Miller and Rumbaugh demonstrating the memory-rescuing potential of inhibiting histone deacetylases (HDACs), a family of signaling enzymes that act like molecular switches, silencing gene expression by controlling access to the cell’s nuclear cache of tightly compacted DNA. Mutations in HDACs genes have been associated with health problems including cancer, inflammatory and autoimmune diseases, metabolic disorders and loss of memory function. Miller and Rumbaugh note that current efforts by many research teams focus on developing “isoform-selective” HDAC inhibitors—for example, select members of Class 1 HDACs such as HDAC 1, -2 or -3—in order to limit the potential for unwanted side effects. However, the Scripps Florida researchers wondered if some of the potential of memory rescue could be lost with this increased selectivity.

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To investigate, in the current study, the Scripps Florida team used inhibitors initially developed by Professor Joel Gottesfeld, a molecular biologist on TSRI’s La Jolla campus, and subsequently by biotech firm Repligen Corporation, to attack more than one form of Class 1 HDAC at the same time in a mouse model of Alzheimer’s disease. “We wanted to find out which inhibitors were the most selective and the most effective in restoring memory function,” Miller said. “We found the key to memory restoration was the growth of new synapses (synaptogenesis), which required simultaneous inhibition of multiple HDACs.” “We found evidence that better synapse growth was associated with less specific inhibition of Class 1 HDACs,” Rumbaugh added. “There was a clear correlation between synapse building— which may lead to improved network power—and memory restoration by the different HDAC inhibitors.” Interestingly, memory was not enhanced in normal animals by chronic pretreatment with multiple HDAC inhibitors, suggesting a diseased brain responds to these compounds differently than a healthy brain. In addition to Miller and Rumbaugh, who was first author the study with Research Associate Stephanie E. Sillivan, other authors of the study, “Pharmacological Selectivity within Class I Histone Deacetylases Predicts Effects On Synaptic Function and Memory Rescue,” include Emin D. Ozkan, Camilo S. Rojas, Christopher R. Hubbs, Massimiliano Aceti and Sathyanarayanan V. Puthanveettil of TSRI; Mark Kilgore and J. David Sweatt of The University of Alabama at Birmingham; and Shashi Kudugunti and James Rusche of Repligen Corporation. For more information on the study, See http://www.nature.com/npp/journal/vaop/ncurrent/full/npp201593a.html The work was supported by the National Institute on Drug Abuse (R01DA034116; R03DA033499), National Institute for Neurological Disorders and Stroke (R01NS064079; R21NS082640); National Institute for Mental Health (R01MH096847; R01MH57014); and Repligen Corporation. Scripps Florida Scientists Show Antitumor Agent Can Be Activated by Natural Response to Cell Stress Findings Point to New Therapy Against Prostate and Other Cancers Scientists from the Florida campus of The Scripps Research Institute (TSRI) have found that a drug candidate with anticancer potential can be activated by one of the body’s natural responses to cellular stress. Once activated, the agent can kill prostate cancer cells. “There is no proven drug right now with these activities,” said Ben Shen, vice chair of TSRI’s Department of Chemistry and senior author of the new study, “so this points the way toward a new therapeutic opportunity.” The study, published recently by the journal Proceedings of the National Academy of Sciences, highlights the potential of the natural compound called leinamycin (LNM) E1 for

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development as a “prodrug,” a medication converted through a metabolic process in the body to become an active therapy. Shen’s research has focused on developing natural products into potential therapies. As part of this effort, he heads the Natural Products Initiative at TSRI, a library available for screening with 500 pure natural products, 2,000 fractions, and 7,500 crude extracts, prepared from 4,000 Actinomycetals. Among these are “antitumor antibiotics” like LNM, which are produced by species of the soil dwelling bacterium Streptomyces and are known to impede cancer cell growth and multiplication. Some antitumor antibiotics are already in use as chemotherapy agents. In the new study, the Scripps Florida team collaborated with scientists at the University of Wisconsin, Madison to examine whether LNM E1 can be activated by reactive oxygen species, which are naturally occurring molecules containing oxygen that play essential roles in cell signaling. During times of stress, levels of reactive oxygen species can rise significantly and may trigger apoptosis or programmed cell death. It is now widely accepted that many cancer cells are, by their very nature, under high oxidative stress. The results were promising. “Our study shows unambiguously that when LNM E1 is activated by cellular reactive oxygen species, it causes DNA damage and cell death in cancer cells,” said Ming Ma, co-first author of the study with Sheng-Xiong Huang. The team further demonstrated the therapeutic potential of LNM E1 by showing it to be effective against two prostate cancer cell lines, which are known to exist under high oxidative stress and with increased levels of reactive oxygen species. The study also reveals critical new insights into LNM biosynthesis, setting the stage to tailor intermediate steps in the creation of new LNM analogues. In addition to Shen, Ma and Huang, other authors of the study, “Leinamycin E1 Acting as an Anticancer Prodrug Activated by Reactive Oxygen Species,” include Dong Yang and Jeremy R. Lohman of TSRI; Bong-Sik Yun, Gudrun Ingenhorst, Yong Huang, Hirak S. Basu, Dawn R. Church, Gong-Li Tang, Jianhua Ju and George Wilding of the University of Wisconsin-Madison. The work was supported in part by the National Institutes of Health (grant CA106150). New Study Brings Together Neuroscience and Psychology to Paint More Complete Picture of Sleep and Memory In Macbeth, Shakespeare describes sleep as “the death of each day’s life,” but he may have gotten it wrong. Sleep, as it turns out, may be the one thing that keeps our memories alive and intact. A new study from the Florida campus of The Scripps Research Institute (TSRI) integrates neuroscience and psychological research to reveal how sleep is more complex than the Bard might

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have imagined. The new research, published online ahead of print by the journal Cell, shows in animal models that sleep suppresses the activity of certain nerve cells that promote forgetting, insuring that at least some memories will last. “Many scientists have tried to figure out how we learn and how our memories become stabilized,” said Ron Davis, chair of the TSRI Department of Neuroscience and senior author of the study. “But far less attention has been paid to forgetting, which is a fundamental function for the brain and potentially has profound consequences for the development of memory therapeutics. Our current study merges the neuroscience of forgetting, that is, the brain mechanisms that lead to forgetting, and the psychology of forgetting into an integrated picture.” Early studies from psychology suggest that sleep facilitates memory retention by stopping interference caused by mental and behavioral activity. That is, sleep essentially isolates the brain from all of the stimuli that can interfere with memory storage. Neuroscience research, on the other hand, suggests that sleep facilitates memory retention by enhancing memory stability or what is called consolidation. The new study in experimental animals reveals the biological underpinnings of the earlier psychology studies, pointing to the activity of the neurotransmitter dopamine. Dopaminergic activity is known to regulate various types of “plasticity”—the ability of the brain to change in direct response to learning and memory formation. That ability includes forgetting as well. The study shows that increasing sleep, with either a sleep-promoting drug or by genetic stimulation of the neural sleep circuit, decreases signaling activity by dopamine, while at the same time enhancing memory retention. Conversely, increasing arousal stimulates dopamine signaling and accelerates forgetting. This signal activity isn’t constant but is tied directly to the animal’s arousal level. “Our findings add compelling evidence to support the model that sleep reduces the forgetting signal in the brain, thereby keeping memories intact,” Davis said. “As sleep progresses to deeper levels, dopamine neurons become less reactive to stimuli and this leads to more stable memories. While the findings bolster earlier psychological studies, they are also not incompatible with more recent findings in neuroscience. The authors note the effects of sleep on memory consolidation and forgetting may operate in parallel and independently of one another or, more intriguingly, in serial in a dependent fashion, with reduced forgetting a prerequisite for sleep-facilitated consolidation. “We all know that sleep helps us remember,” said Research Associate Jacob A. Berry, the first author of the study and a member of the Davis lab. “Importantly, we have revealed that one of the ways sleep protects a new memory is by quieting dopamine neuron activity that causes forgetting. Since laboratory animals and humans share a need for sleep, as well as many genetic and circuit mechanisms underlying learning and memory, our findings may shed light on the mechanisms underlying the interaction between sleep and memory in humans.”

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In addition to Davis and Berry, other authors of the study, “Sleep Facilitates Memory by Blocking Dopamine Neuron-Mediated Forgetting,” are Isaac Cervantes-Sandoval and Molee Chakraborty of TSRI. This work was supported by the National Institutes of Health (grants R37 NS19904 and R01 NS051251). Scripps Florida Scientists Uncover Unique Role of Nerve Cells in the Body’s Use of Energy While it is well-known that weight gain results from an imbalance between what we eat and our energy expenditure, what is not obvious is the role that the nervous system plays in controlling that energy balance. Now scientists from the Florida campus of The Scripps Research Institute (TSRI) have shed light on that question. “Our new study has identified novel populations of nerve cells that regulate appetite, thermogenesis and physical activity,” said TSRI Professor Baoji Xu, who led the research. “We think these neurons could be targets for drug development.” The findings were published by the journal Cell Metabolism online ahead of print on June 11. In the new study, Xu and his colleagues examined several groups of neurons that express a substance called “brain-derived neurotrophic factor” (BDNF) within a small brain region called the paraventricular hypothalamus. BDNF is an extremely important protein in the brain and is involved in a number of functions. It has been shown that deleting the BDNF gene causes significant problems, among them, dramatically increased appetite (hyperphagia) and severe obesity. The new study shows that deleting the BDNF gene also impairs thermogenesis—the ability of cells to burn fat to produce heat. The study further reveals two distinct types of BDNF neurons—those that control appetite or satiety and those that control thermogenesis. Not only do these two groups play different biological roles, they are located in two separate sections of the paraventricular hypothalamus brain region. This “geographical” split raises some interesting questions. “We don’t yet know what the distinctive placement means to the control of body weight, nor do we know if these two clusters of neurons communicate with each other as yet,” said Juanji An, the first author of the study and a member of the Xu lab. “But given the fact that mice and humans with mutations in the BDNF gene or its receptor develop severe obesity, a better understanding of the mechanism underlying the effect BDNF has on body weight could provide great insights into the regulation of energy balance.” Xu is also hopeful about the potential of BDNF as a drug target. “Our findings suggest that activation of each of these two populations of neurons should powerfully suppress appetite or promote energy expenditure,” he said. However, he cautions that because BDNF’s functions are so

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widespread, a good drug candidate would need to closely target only BDNF-expressing neurons in the paraventricular hypothalamus, thus limiting potential side effects. In addition to Xu and An, other authors of the study, “Discrete BDNF Neurons in the Paraventricular Hypothalamus Control Feeding and Energy Expenditure,” include Guey-Ying Liao and Clint E. Kinney of TSRI, and Niaz Sahibzada of the Georgetown University Medical Center. See http://www.cell.com/cell-metabolism/home The work was supported by grants from the National Institutes of Health (DK089237 and DK103335) and the Klarman Foundation. Scripps Florida Scientists Identify a Potential New Treatment for Osteoporosis Scientists from the Florida campus of The Scripps Research Institute (TSRI) have identified a new therapeutic approach that, while still preliminary, could promote the development of new bone-forming cells in patients suffering from bone loss. The study, published recently in the journal Nature Communications, focused on a protein called PPARy (known as the master regulator of fat) and its impact on the fate of stem cells derived from bone marrow (“mesenchymal stem cells”). Since these mesenchymal stem cells can develop into several different cell types—including fat, connective tissues, bone and cartilage—they have a number of potentially important therapeutic applications. The scientists knew that a partial loss of PPARy in a genetically modified mouse model led to increased bone formation. To see if they could mimic that effect using a drug candidate, the researchers combined a variety of structural biology approaches to rationally design a new compound that could repress the biological activity of PPARy. The results showed that when human mesenchymal stem cells were treated with the new compound, which they called SR2595 (SR=Scripps Research), there was a statistically significant increase in osteoblast formation, a cell type known to form bone. “These findings demonstrate for the first time a new therapeutic application for drugs targeting PPARy, which has been the focus of efforts to develop insulin sensitizers to treat type 2 diabetes,” said Patrick Griffin, chair of the Department of Molecular Therapeutics and director of the Translational Research Institute at Scripps Florida. “We have already demonstrated SR2595 has suitable properties for testing in mice; the next step is to perform an in-depth analysis of the drug’s efficacy in animal models of bone loss, aging, obesity and diabetes.” In addition to identifying a potential new therapeutic for bone loss, the study may have even broader implications. “Because PPARG is so closely related to several proteins with known roles in disease, we can potentially apply these structural insights to design new compounds for a variety of therapeutic applications,” said David P. Marciano, first author of the study, a recent graduate of TSRI’s PhD

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program and former member of the Griffin lab. “In addition, we now better understand how natural molecules in our bodies regulate metabolic and bone homeostasis, and how unwanted changes can underlie the pathogenesis of a disease.” Marciano will focus on this subject in his postdoctoral work in the Department of Genetics at Stanford University. In addition to Marciano and Griffin, other authors of the study, “Pharmacological Repression of PPARγ Promotes Osteogenesis,” are Dana S. Kuruvilla, Siddaraju V. Boregowda, Alice Asteian, Travis S. Hughes, Ruben D. Garcia-Ordonez, Scott J. Novick, Cesar A. Corzo, Tanya M. Khan, Douglas J. Kojetin, Donald G. Phinney and Theodore M. Kamenecka of TSRI; and John B. Bruning of The University of Adelaide (Australia). See http://www.nature.com/ncomms/index.html The work was supported by the National Institutes of Health (grants DK08026, MH084512, OD018254-01, DK097890, DK103116 and DK101871). Scripps Florida Study Points to Drug Target for Huntington’s Disease Huntington’s disease attacks the part of the brain that controls movement, destroying nerves with a barrage of toxicity, yet leaves other parts relatively unscathed. Scientists from the Florida campus of The Scripps Research Institute (TSRI) have established conclusively that an activating protein, called “Rhes,” plays a pivotal role in focusing the toxicity of Huntington’s in the striatum, a smallish section of the forebrain that controls body movement and is potentially involved in other cognitive functions such as working memory. “Our study definitively confirms the role of Rhes in Huntington’s disease,” said TSRI Assistant Professor Srinivasa Subramaniam, who led the study. “Our next step should be to develop drugs that inhibit its action.” The study was published recently online ahead of print by the journal Neurobiology of Disease. In an earlier study, Subramaniam and his colleagues showed that Rhes binds to a series of repeats in the huntingtin protein (named for its association with Huntington’s disease), increasing the death of neurons. The new study shows deleting Rhes significantly reduces behavioral problems in animal models of the disease. In addition, the study took the research further and revealed the effects of adding Rhes to the cerebellum, a brain region normally not affected in Huntington’s. Remarkably, Huntington disease animals injected with Rhes experienced an exacerbation of motor issues, including loss of balance and coordination. Subramaniam and his colleagues also found lesions and damaged neurons in the cerebellum, confirming Rhes is sufficient to promote toxicity and showing that even those regions of the brain normally impervious to damage can become vulnerable if Rhes is overexpressed.

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“Perhaps the biggest question to emerge from this study is whether Rhes is a good drug target for Huntington’s disease,” Subramaniam said. “The short answer is ‘yes.’ Drugs that disrupt Rhes could alleviate Huntington’s pathology and motor symptoms.” “Many Huntington’s disease patients experience psychiatric-related problems, such as depression and anxiety,” added Supriya Swarnkar, the first author of the study and a member of Subramaniam’s lab. “But it’s unclear whether they are the cause or consequences of the disease. We think, by targeting Rhes, we might block the initiation of Huntington’s, which we predict would afford protection against psychiatric-related problems as well.” In addition to Swarnkar and Subramaniam, other authors of the study, “Ectopic Expression of the Striatal-enriched GTPase Rhes Elicits Cerebellar Degeneration and an Ataxia Phenotype in Huntington Disease,” are Youjun Chen, William Pryor, Neelam Shahani and Damon Page of TSRI. See http://www.sciencedirect.com/science/article/pii/S0969996115001850 The work was supported by the State of Florida. Small RNAs Found to Play Important Roles in Memory Formation Scientists from the Florida campus of The Scripps Research Institute (TSRI) have found that a type of genetic material called “microRNA” plays surprisingly different roles in the formation of memory in animal models. In some cases, these RNAs increase memory, while others decrease it. “Our systematic screen offers an important first step toward the comprehensive identification of all miRNAs and their potential targets that serve in gene networks important for normal learning and memory,” said Ron Davis, chair of TSRI’s Department of Neuroscience who led the study. “This is a valuable resource for future studies.” The study was published in the June 2015 edition of the journal Genetics. Unlike some types of RNA, microRNAs (miRNAs) do not code for proteins but instead regulate various biological processes by modulating the level of gene expression. A number of studies have shown that miRNAs are critical for normal development and cellular growth and may contribute to the complexity of neurodegenerative diseases. In the new study, 134 different miRNAs were tested for roles in learning and memory in the central nervous system of Drosophila melanogaster, the common fruit fly, which is a recognized animal model for memory studies. The researchers tested the potential involvement of miRNAs in intermediate-term memory by silencing them individually and identified at least five different miRNAs involved in memory formation or retention. “Among the five miRNAs identified in this study, we found one that is necessary for memory formation,” said Research Associate Germain U. Busto, a first author of the study with Research

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Associate Tugba Guven-Ozkan. “Interestingly, its human counterpart is altered in several neurodegenerative diseases, including Alzheimer’s and Huntington’s. It’s possible that this might be a potential model to study and solve some specific aspects of those disorders.” Surprisingly, the researchers found some miRNAs decreased memory formation, while others increased it. The identified miRNAs affected either neuronal physiology underlying memory formation or the development of the nervous system. “These microRNAs are highly regulated during brain development and for adult brain function,” said Guven-Ozkan. “When misregulated, they may exacerbate brain diseases like autism, and Alzheimer’s and Huntington’s diseases. We’d like to pinpoint learning and memory pathways to understand how they may lead to human disease.” In addition to Davis, Busto and Guven-Ozkan, other authors of the study, “microRNAs That Promote or Inhibit Memory Formation in Drosophila melanogaster,” include Tudor A. Fulga and David Van Vactor of Harvard Medical School. For more information, seehttp://www.genetics.org/content/200/2/569.abstract. The work was supported by National Institutes of Health (grants R37 NS19904 and R01 NS069695).

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PART II – MILESTONES AND UPDATES FROM AGREEMENT SECTIONS

Part II of the annual report addresses the milestones and provides updates to specific components of the formal agreement, listed by agreement section. Section 9.3 Annual Report. Scripps (The Scripps Research Institute) shall prepare

the Annual Report for Scripps Florida each year and deliver such Annual Report to Funding by August 31st of each year. The Annual Report shall include, but not be limited to, the following information:

Section 9.3(a) An accounting of the expenditures of Grant Funds for the twelve months

ended June 30th of each year (the "Report Year" [as amended]) and financial commitments made by Scripps during the Report Year.

Report of SFFC Grant Fund Cash Disbursements from October 1, 2014 to June 30, 2015.

Salaries & Benefits $ 7,236,761 Supplies $ 88,450 Scientific equipment $ 1,878,941 External affairs & other program support $ 2,062,733 Project commencement, facilities, administration and other capital expenditures $ 5,788,648 Total $ 17,055,533

The schedule reflects cash expenditures charged to the grant from the State of Florida from October 1, 2014 to June 30, 2015. The expense categories set forth above reflect those used by Scripps to report grant activity to grantors. This schedule excludes unspent grant funds received of $73,791,168 restricted to support Scripps Florida. There were no new financial commitments made during the report year.

Section 9.3(b) Data regarding the activities and performance of Scripps Florida during

such Report Year and detailing the progress of Scripps in meeting its Business Plan, including but not limited to:

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Section 9.3(b)i Information on the number and salary level of jobs created by Scripps within Scripps Florida, including the number and salary level of jobs created for residents of Florida;

On June 30, 2015, Scripps Florida employed 585 people. Of those, 521 were full-time. The breakdown of those full-time employees is shown below.

Faculty 49Research Faculty 7Staff Scientists 18Research Associates 174Scientific Support 117Administrative Support 156Total 521

In addition, on June 30, 2015 Scripps Florida employed an additional 64 employees who were part-time, research interns, or summer interns, for a total employee population of 585. Of the 205 employees hired between July 1, 2014 and June 30, 2015, 66 were residents of Florida and 45 were residents of Palm Beach County. The average salary/range for those employees was: Faculty $33,280 - $412,548 Research Faculty $125,382 - $177,611 Staff Scientists $62,566 - $100,006 Research Associates $37,000 - $75,213 Administrative Support $60,112 average

Section 9.3(b)ii A description of the status of the performance expectations set forth in Section 9.5 of this Agreement and the disbursement conditions set forth in Schedule 4.4(c) of this Agreement;

See responses to Sections 9.5 and 4.4(c), below. Section 9.3(b)iii Information on the positions and funds required to be committed for

equipment for such positions by means of the next annual disbursement of Grant Funds; All grant funds have been disbursed. That said, the Scripps Research Institute will submit the budgets for Scripps Florida for the year ending September 30, 2015, to SFFC after approval by the Scripps Research Institute Board of Trustees expected to occur on September 20, 2015. These budgets set forth all anticipated revenue and expenses for Scripps Florida for the next fiscal

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year. The equipment requirements for new positions will be incorporated into these budgets.

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Fiscal year equipment reporting:

Approximately $ 1,047,090 of equipment – acquired with State grant funds was purchased from October 1, 2014 through June 30, 2015. In addition, $ 3,876,163 of equipment was purchased using non SFFC funds during the period from October 1, 2014 through June 30, 2015.

Section 9.3(b)iv Commencing with the Annual Report for 2006 Report Year and ending

with the Report Year after which Scripps has moved the Scripps Florida operations to its permanent facility and such facility is fully operational, a description of the status of Scripps’ relocation to its second planned temporary facility and the progress of construction activities for its permanent facility, as described in the Business Plan, including a projected date for and status of Scripps’ occupancy of its permanent facility.

Scripps Florida officially opened its Permanent Facilities in February of 2009. Section 9.3(b)v And commencing with the Annual Report for the Report Year during

which Scripps commences activities at its permanent facility, a description of the status of Scripps' activities in its permanent facility, including its educational and outreach programs.

Over the past eleven years (since 2004) Scripps Florida has placed considerable effort in community and education outreach programs. Palm Beach County K-12 students and teachers have participated in science education lessons and events designed and presented by Scripps Florida education outreach, graduate students, post doctoral fellows, faculty and staff researchers. The programs described in Sections 4.4(c)6 and 4.4(c)7 define the goals of Scripps Florida’s K-12 education programs: to work directly with students and teachers, to help develop instructional materials, and to contribute to science literacy in Palm Beach County and the State of Florida. Scripps Florida has taken a leadership role in science education since its inception; presenting at state and national meetings such as National Science Teacher’s Association, American Association for the Advancement of Science and the International Teacher/Scientist Conference. In Florida, education outreach has participated in education meetings with the Florida State Department of Education, Florida Council of 100, State University System of Florida Board of Governors, STEM Florida and the Sunshine State Scholars. To date, more than twenty thousand students, teachers, and community members of Palm Beach County have participated in the Scripps Florida Education Outreach programs. Scripps Florida community outreach has offered opportunities for the public to gain insight into cutting edge biomedical research while providing opportunities for Scripps Florida faculty and staff to respond to the social needs of Palm Beach County. Since 2004, Scripps Florida has hosted and/or participated in Community Outreach events that include symposia on Alzheimer’s disease, breakthroughs in cancer research, and current discoveries in drug development, just to name a few.

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Following is a list of Scripps Florida Education and Community outreach activities from the past year:

Community Outreach

Date Participants Recipients/Event

Community Outreach 18-Sep-14 Roy Smith NuVista Community Outreach

14-Jul-14 Niedernhofer Women's Foundation of Palm Beach County Meeting

Community Outreach 15-Aug-14 Niedernhofer

Scripps tour for American Association of University Women of Palm Beach County

Community Outreach 3-Sep-14 Paul Robbins Dinner with June Jones Community Outreach 23-Sep-14 Thomas Kodadek Frenchman's Creek - Cancer Biology Community Outreach 1-Oct-14 Thomas Kodadek Focus Group Meeting with Margaret

Wilesmith Community Outreach 2-Oct-14 Thomas Kodadek Moments of Change Conference at The

Breakers Community Outreach 6-Oct-14 Thomas Kodadek Kravis Center Forum Club Community Outreach 6-Oct-14 Thomas Kodadek Homeless Coalititon Event Community Outreach

08-Oct-14 Niedernhofer Lunch & Scripps Tour with Philanthropy guest Diane Trout

Community Outreach 16-Oct-14 Smith, Miller, Robbins, Niedernhofer

Philanthrophy Event: BMO/Harris Bank - Aging Presentations

Community Outreach 16-Oct-14 Niedernhofer BMO Cocktail Reception and tour of Scripps Community Outreach 17-Oct-14 Thomas Kodadek LPBC Bowl-a-thon Community Outreach 20-Oct-14 Thomas Kodadek Palm Beach Civic Association meeting Community Outreach 21-Oct-14 Paul Robbins District 18 Congressional Debate Community Outreach 24-Oct-14 Thomas Kodadek Leadership Palm Beach County Community Outreach 30-Oct-14 Thomas Kodadek Keynote speaker at Ballenisles LGA

Welcome Back Lunch

Community Outreach 3-Nov-14 Ron Davis Meeting and Tour with Community Foundation (D. Houston), Scripps Florida

Community Outreach 10-Nov-14 Niedernhofer Scripps tours for philanthropy Community Outreach 11-Nov-14 Thomas Kodadek Frenchman's Creek Cancer/ALS Event

Community Outreach 11-Nov-14 Matt Disney Night at Frenchmen's Creek Community Outreach 13-Nov-14 Thomas Kodadek Address North County Development Board

Meeting

Community Outreach 13-Nov-14 Ron Davis Community Foundation presentation at Palm Beach Yacht Club, West Palm Beach, FL

Community Outreach 1-Dec-14 Kirill Martemyanov, Sathya Puthanveettil,

Gavin Rumbaugh

O'Keefe Symposia

Community Outreach 3-Dec-14 Ron Davis Alzhiermer's presentation at Palm Beach Yacht Club, West Palm Beach, FL

Community Outreach 5-Dec-14 Thomas Kodadek Leadership Palm Beach County Community Outreach 9-Dec-14 Paul Robbins Philanthropy Lunch & Learn Presentation Community Outreach 10-Dec-14 Thomas Kodadek Frenchman's Creek Lab tour and lunch Community Outreach

11-Dec-14 Niedernhofer "Smith College Club", EastPointe Country Club, Jupiter, FL Guest speaker

Community Outreach 17-Dec-14 Christoph Rader Klorfine Foundation, Scripps Florida Campus

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Community Outreach 29-Dec-14 Thomas Kodadek Venture Meeting Community Outreach 5-Jan-15 Thomas Kodadek BCG interview on cancer immunotherapy and

innovation waves in oncology

Community Outreach 7-Jan-15 Thomas Kodadek PGA National Forum Community Outreach 12-Jan-15 Thomas Kodadek Economic Forum of PBC at the Kravis Community Outreach 13-Jan-15 Ron Davis Alzheimer presentation at Mandel Jewish

Community Center, Palm Beach Gardens, FL

Community Outreach 14-Jan-15 Ron Davis Ryan Licht Sang Symposium: Onset Bipolar Medical Briefing Luncheon, Palm Beach, FL

Community Outreach 14-Jan-15 Niedernhofer Hunters Run TSRI Tour Community Outreach 16-Jan-15 Thomas Kodadek Leadership Palm Beach County meeting Community Outreach 20-Jan-15 Roy Smtih Philanthrophy: Women of Vision PNC Event Community Outreach 22-Jan-15 Thomas Kodadek LPBC Reception at Scripps Community Outreach 22-Jan-15 Matt Disney Leadership Palm Beach County Community Outreach 23-Jan-15 Thomas Kodadek Hosted Frenchman's Creek Women's Cancer

Association dinner

Community Outreach 28-Jan-15 Scripps Faculty and Philanthropy

CELLebrate Science Reception, Gardens Mall, Palm Beach Gardens, FL

Community Outreach 28-Jan-15 Niedernhofer

Women's Foundation of Palm Beach County Meeting

Community Outreach 31-Jan-15 Campus Wide Scripps Florida, CELLebrate Science Day, Gardens Mall

Community Outreach 1-Feb-15 Kirill Martemyanov Neuroscience Symposia Community Outreach

01-Feb-15 Ja, Kishi,

Niedernhofer "CELLebrate Science with Scripps Florida" event at The Gardens Mall

Community Outreach 3-Feb-15 Hyeryun Choe Infectious Diseases in Children, NJ Community Outreach 3-Feb-15 Ron Davis Meeting and Tour with Community

Foundation (D. Hanley), Scripps Florida

Community Outreach 10-Feb-15 Thomas Kodadek Meeting at Lewis Center-Food for Homeless Community Outreach 12-Feb-15 Thomas Kodadek PGA Women's Cancer Awareness Days

Luncheon

Community Outreach 15-Feb-15 Kirill Martemyanov, Sathya Puthanveettil

O'Keefe Symposia

Community Outreach 19-Feb-15 Thomas Kodadek Greenwich CT Public radio station interview regarding BioMedical Research

Community Outreach 21-Feb-15 Thomas Kodadek Black Ties & French Fries in Wonderland-benefiting young children impacted by abuse & violence.

Community Outreach 22-Feb-15 Thomas Kodadek Hosted Families First dinner Community Outreach 26-Feb-15 Thomas Kodadek Presentation to the Central Palm Beach

County Chamber of Commerce

Community Outreach 26-Feb-15 Ron Davis, Damon Page, Brock Grill

O'Keeffe Neuroscience Symposium

Community Outreach 26-Feb-15 Niedernhofer

Scripps tour for American Association of University Women of Palm Beach County

Community Outreach 27-Feb-15 Thomas Kodadek Frenchman's Creek Community Outreach 9-Mar-15 Thomas Kodadek Palm Beach State College Congressional

Awards Community Outreach 10-Mar-15 Courtney Miller The Society of the Four Arts, Guest Speaker

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Community Outreach 11-Mar-15 Ron Davis Mass General Alzheimers Presentation - Lake Pavillion, West Palm Beach, FL

Community Outreach 12-Mar-15 Patrick Griffin Presentation/discussion on diabetes presented to Mandel JCC

Community Outreach 15-Mar-15 Kirill Martemyanov, Sathya Puthanveettil

Carol Mostad Group

Community Outreach 15-Mar-15 Kirill Martemyanov Northern Trust Event Community Outreach 15-Mar-15 Gavin Rumbaugh Society for the 4 Arts Presentation Community Outreach 16-Mar-15 Damon Page Autism Speaks Town Hall Meeting, Palm

Beach Country Club, Palm Beach, FL

Community Outreach 21-Mar-15 Mark Sundrud Speaker - Crohn's & Colitis Foundation education program, Jupiter Medical Center, Jupiter, FL

Community Outreach 22-Mar-15 Ron Davis and Roy Smith

Scripps Science will make 80 the new 50 at Palm Beach Country Club, Palm Beach, FL

Community Outreach 25-Mar-15 Ron Davis, Damon Page, Brock Grill

O'Keeffe Neuroscience / Northern Trust March Session Neurological Issues presentation, Scripps Florida

Community Outreach 31-Mar-15 Thomas Kodadek Meeting w/ Margaux's Miracle Foundation about Childhood Ewing's Sarcoma Research

Community Outreach 31-Mar-15 Damon Page Autism Event at Palm Beach Yacht Club, West Palm Beach, FL

Community Outreach 1-Apr-15 Ron Davis, Damon Page

O'Keeffe Neuroscience Symposium Dinner "Brain Health: Understanding Brain Disorders", Scripps Florida

Community Outreach 01-Apr-15 Niedernhofer TSRI Aloha event Community Outreach 10-Apr-15 Seth Tomchik Palm Beach Mental Health Association

Lunch and Learn

Community Outreach 14-Apr-15 Ron Davis Sea Colony Tour, Scripps Florida Community Outreach 14-Apr-15 Ron Davis and Roy

Smith Quantum Foundation, Scripps Florida

Community Outreach 15-Apr-15 Thomas Kodadek Presentation to the Jewish Community Center Community Outreach 16-Apr-15 Thomas Kodadek Families First Community Outreach 18-Apr-15 Roy Smith Prader Willi Florida Chapter, Invited Speaker Community Outreach 23-Apr-15 Paul Robbins Meeting with June Jones at Scripps Community Outreach 24-Apr-15 Thomas Kodadek LPBC Excellence Award Community Outreach 24-Apr-15 Gill PNC Food for Thought Lunch Series Community Outreach 28-Apr-15 Thomas Kodadek PGA National gift presentation to Scripps

Cancer Biology department

Community Outreach 28-Apr-15 Patrick Griffin, Chakraborty

Diabetes presentation and receptioin @ Scripps Florida

Community Outreach 28-Apr-15 Ja/Niedernhofer Invited Speaker, Palm Beach Yacht Club Community Outreach 28-Apr-15 Paul Robbins Presentation at Palm Beach Yacht Club Community Outreach

05-May-15 Niedernhofer Presenter at the Young Entrepreneurs Academy graduation

Community Outreach 6-May-15 Thomas Kodadek Lewis Center Community Outreach 7-May-15 Thomas Kodadek Presentation to The Nexus Society Community Outreach 11-May-15 Thomas Kodadek BDB Life Sciences & Healthcare Task Force

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Community Outreach 11-May-15 Niedernhofer

Radio Show interview by Scott Greenberg, OMG my mom's getting older and so am I!

Community Outreach 11-May-15 Niedernhofer

FAU Medical School Celebration at the home of Sydell Miller, Palm Beach

Community Outreach 16-May-15 Thomas Kodadek Central Palm Beach Chamber of Commerce Community Outreach 18-May-15 Niedernhofer Vice Mayor Mary Lou Berger tour of Scripps Community Outreach 18-May-15 Paul Robbins Vice Mayor Berger visit Community Outreach

28-May-15 Niedernhofer Attended BioFlorida "Raising Capital for Biotech Ventures in Florida", Scripps-Florida, Jupiter, FL

Community Outreach 3-Jun-15 Ron Davis, Damon Page, Jacob Berry,

William Pryor

Community Foundation at Kravis Center, West Palm Beach, FL

Community Outreach 08-Jun-15 Niedernhofer Videographer/Interview for TSRI Community Outreach 9-Jun-15 Thomas Kodadek Faculty representative for the Community

Relations Group Meeting at Scripps

Community Outreach 10-Jun-15 Thomas Kodadek Address Economic Forum at the Kravis Center

Community Outreach 15-Jun-15 Kirill Martemyanov, Sathya Puthanveettil

Foundations Symposia

Community Outreach 17-Jun-15 Thomas Kodadek North Palm Beach Chamber of Commerce Community Outreach

17-Jun-15 Niedernhofer Business Before Hours - State of the Chamber: Game Changer Edition

Community Outreach 23-Jun-15 Niedernhofer TSRI Tours for Philanthropy Community Outreach 20-Oct-14

27-Oct-14 Ron Davis, Damon Page, Brock Grill,

Youjun Chen

O'Keeffe Neuroscience Symposium

Community Outreach Monthly Niedernhofer

Community Relations Group Scripps FL, Meetings

Community Outreach Monthly Niedernhofer

Scripps Florida representative to the Northern Palm Beach County Chamber, Trustee

Community Outreach Monthly Niedernhofer Faculty Philanthropy Committee Community Outreach

Quarterly Niedernhofer Meeting with Ms. June Jones, Scripps benefactor

Education Outreach Date Participants Recipients/Event Education Outreach Jul-14 to Jun-

15 Courtney Miller Intern/Thesis Advisor to 4 FAU undergraduates

Education Outreach

5-Oct-13 Paul Robbins Thesis committee meeting Jonathon Proto University of Pittsburgh

Education Outreach 11-Jul-14 Seth Tomchik Keith Murphy Committee Meeting - FAU grad program

Education Outreach 23-Jul-14 Dr. Rosie Albarran-Zeckler

Custom lesson for PBSC Summer Youth College Program – Boca Raton Campus

Education Outreach 23-Jul-14 Chakraborty Florida Curriculum Committee Education Outreach

23-Jul-14 Niedernhofer Thesis committee/defense of Katy Wack, University of Pittsburgh School of Medicine graduate program

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Education Outreach 24-Jul-14 Drs. Laura Pedro Rosa, Tamara Boto, and Rosie Albarran-Zeckler; and Deborah Leach-Scampavia and Jennifer Kabis

Science Family – DNA Isolation (El Sol Community Center) Spanish Language

Education Outreach 28-Jul-14 Drs. Sarbani Goshal and Rosie Albarran-Zeckler

Science Path - PBSC'S Teachers Program

Education Outreach 07/28/2014 Niedernhofer

High school intern poster presentation; mentor to Margo Orlen

Education Outreach 30-Jul-14 Damon Page Renaissance Learning Center Board Meeting, West Palm Beach, FL

Education Outreach 08/06/2014 Niedernhofer Mentoring session, Scripps-Florida Education Outreach

08/06/2014 Niedernhofer Scripps Florida Post-doctoral association mentoring session

Education Outreach 7-Aug-14 Courtney Miller Thesis Committee Meeting, Shannon Sanguinetti (FAU Graduate Program)

Education Outreach 13-Aug-14 Dr. Rosie Albarran-Zeckler

Palm Beach County Math Teacher Symposium

Education Outreach 14-Aug-14 Dr. Rosie Albarran-Zeckler, Deborah Leach-Scampavia, and Kristin Lidinsky

Palm Beach County Science Teacher Symposium

Education Outreach 15-Aug-14 Paul Robbins Undergraduate seminars Education Outreach 26-Aug-14 Paul Robbins Toastmasters Humorous Speech Contest Education Outreach 3-Sep-14 Kendall Nettles Invited Presenter: Physiology, Biophysics and

Systems Biology Seminar Series, Weill Cornell Medical College NY

Education Outreach 9-Sep-14 Chakraborty Panel Academic Job Search Education Outreach 10-Sep-14 Paul Robbins Annual Graduate Program dinner Education Outreach 20-Sep-14 Dr. Rosie Albarran-

Zeckler Tiger Woods Foundation Career and College Day for High School Students

Education Outreach 22-Sep-14 Katrin Karbstein The Benjamin School: 3rd Grade Presentation - Yeast as a model organism

Education Outreach 22-Sep-14 Dr. Rosie Albarran-Zeckler and Deborah Leach-Scampavia

Science Family – DNA Isolation (West Palm Beach Community Library)

Education Outreach 9-Oct-14 Courtney Miller Children's Service Council/PBS Panel - Raising America

Education Outreach 10-Oct-14 Matt Disney Discussion with postdoctoral associates on career opportunities

Education Outreach 28-Oct-14 Kendall Nettles Invited Presenter: Nuclear Receptors & Disease, Cold Spring Harbor, NY

Education Outreach 11/2014 Niedernhofer

Teaching in TSRI Graduate Program Investigations in Molecular Biology

Education Outreach 7-Nov-14 Christoph Rader Florida Biomedical Career Symposium, Scripps Florida Campus

Education Outreach 7-Nov-14 Laura Bohn

Florida Biomedical Career Symposium, Jupiter, FL

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Education Outreach 8-Nov-14 Dr. Rosie Albarran-Zeckler, Deborah Leach-Scampavia, and Kristin Lidinsky

Science Family – DNA Isolation (Elliot Museum at Martin County)

Education Outreach 13-Nov-14 Drs. Xin-An Liu, Kymberly Lovell, Jenny Morgenweck, Akaitz Dorronsoro, Heike Fuhrmann-Stroissnigg, Sarbani Goshal, Rosie Albarran-Zeckler; and Deborah Leach-Scampavia

High School Teacher Workshop at Scripps Florida

Education Outreach 25-Nov-14 Brock Grill Ph.D. thesis committee meeting for Scott Baker (University of Minnesota)

Education Outreach 7-Dec-14 Laura Bohn American College of Neuropsychopharmacology Education Outreach 17-Jan-15 Deborah Leach-

Scampavia and Jennifer Kabis , and Drs. Ilaria Drago, Xin-an Liu, Komolitdin Akhmedov, and Rosie Albarran-Zeckler

Students from Palm Beach County Title I schools / Neuroscience Saturday

Education Outreach 26-Jan-15 Graduate students Sany Hoxha, Walter Rogal, Zachary J. Tickner

High School Students/Career panel at Scripps

Education Outreach 29-Jan-15 Matt Disney Graduate Faculty Town Hall Meeting Education Outreach 30-Jan-15 Dr. Matthew Pipkin,

Deborah Leach Scampavia, and Rosie Albarran-Zeckler

Students from 5th and 6th grades / Rosarian Academy – Career Day (Invitation made through Dr. Pipkin)

Education Outreach 14-Feb-15 Paul Robbins Grad recruiting brunch Education Outreach 18-Feb-15 Dr.Matt Gill, Deborah

Leach-Scapavia, and Rosie G. Albarran-Zeckler

Jerry Thomas Elementary School – Science Fair

Education Outreach 21-Feb-15 Deborah Leach-Scapavia and Rosie G. Albarran-Zeckler

Inlet Grove Community High School – Careers in medical field day

Education Outreach 24-Feb-15 Brock Grill Visit to Loxahatchee High School C. elegens course in Loxahatchee, FL

Education Outreach 26-Feb-15 Damon Page Renaissance Learning Center Board Meeting, West Palm Beach, FL

Education Outreach 1-Mar-15 Tina Izard Development of Weiss Middle School 2016 School Year Science and Lab curriculum

Education Outreach 7-Mar-15 Dr. Brian Paegel, Marie Maolne, Wes Conchrane, Jennifer Kabis, Rosie Albarran-Zeckler

Teachers/ Dr. Paegel's Microscopy Teacher Workshop – Beta testing Day 1

Education Outreach 15-Mar-15 Sathya Puthanveettil Mandel Public Library Lectures Education Outreach 17-Mar-15 Katrin Karbstein FIU Undergraduate Research Conference -

Quality Control Mechanisms in Ribosome

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Activity, Miami, FL

Education Outreach 17-Mar-15 Cheryl Marra and Rosie Albarran-Zeckler

Undergraduates/ Florida International University Research Undergraduate Conference

Education Outreach 1-Apr-15 Dr. Tina Izard, Deborah Leach-Scampavia and Rosie G. Albarran-Zeckler

Middle school students/ Weiss School – Dr. Izard’s outreach project

Education Outreach 16-Apr-15 Rosie Albarran-Zeckler and Jennifer Kabis

Leadership Youth Visit to SF

Education Outreach 17-Apr-15 Rosie Albarran-Zeckler and Jennifer Kabis

Undergraduate Students/ Visit from PBSC North Campus

Education Outreach 04/22/2015 Niedernhofer

Teaching in TSRI Graduate Program Cancer Biology

Education Outreach 30-Apr-15 Damon Page Renaissance Learning Center Board Meeting, West Palm Beach, FL

Education Outreach 5-May-15 Thomas Kodadek Education Foundation Education Outreach 8-May-15 Baoji Xu Chao Chen Thesis Defense at Georgetown

University, Washington, DC

Education Outreach 16-Jun-15 Courtney Miller Thesis Committee External Examiner, Veronica Peschansky (Univ of Miami Medical School Graduate Program)

Education Outreach 23-Jun-15 Dr. Pedro Reis Rodrigues, Rosie Albarran-Zeckler, Jennifer Kabis

PBSC's MSI Program participants/ Visit to SF and lab tour

Education Outreach 25-Jun-15 Damon Page Renaissance Learning Center Board Meeting, West Palm Beach, FL

Education Outreach 30-Jun-15 Rosie Albarran-Zeckler and Dr. Heike Fuhrmann-Stroissnigg

Girls Excelling in Math and Science at Science Center in WPB (Speaker: Dr. Heike Fuhrman)

Education Outreach 01/2015-04/2015

Ja Mentor Shivani Patel, Undergraduate Intern, Harriet L. Wilkes Honors College/FAU

Education Outreach 01/2015-Present

Niedernhofer Mentor Clayton Sims, Intern, Harriet L. Wilkes Honors College/FAU

Education Outreach 01/2015-Present

Niedernhofer Mentor Rachael Candela, Intern, Harriet L. Wilkes Honors College/FAU

Education Outreach 01/2015-Present

Ja Mentor Diana Singkornrat, Intern, FAU

Education Outreach 03/2015-06/2015

Niedernhofer Mentor Jolanta Czerwinska, Graduate Student Intern, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland

Education Outreach 03/2015-08/2015

Niedernhofer Mentor Junaid Raya, Intern, Harriet L. Wilkes Honors College/FAU

Education Outreach 05/2013-05/2015

Niedernhofer Teaching in Frontiers in Aging and Regeneration Research, Woods Hole, MA

Education Outreach 05/2015-07/2015

Niedernhofer Mentor Margo Orlen, Intern, Spanish River Community High School

Education Outreach 05/2015-08/2015

Niedernhofer Mentor Sanjay Chandrasekhar, Undergraduate Intern, University of Pennsylvania

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Education Outreach 05/2015-08/2015

Ja Mentor Keuri Reis Santos, Intern, Universidade Federal do ABC, Sao Bernardo do Camp, São Paulo, Brasil

Education Outreach 05/2015-Present

Niedernhofer Mentor Jamie Harris, Intern, Atlantic High School

Education Outreach 05/2015-Present

Ja Mentor Margaux Ehrlich, Intern, Harriet L. Wilkes Honors College/FAU

Education Outreach 07/2014-present

Ja Mentor Keith Murphy, Graduate Student, FAU

Education Outreach 08/2014-05/2015

Niedernhofer Mentor Srigita Madiraju, Intern, Harriet L. Wilkes Honors College/FAU

Education Outreach 08/2014-Present

Niedernhofer Mentor Danielle Hennessy-Wach, Intern, Harriet L. Wilkes Honors College/FAU

Education Outreach 09/2012 to present

Kishi Mentor Delacia Ingram, Intern/Lab Helper, FAU

Education Outreach

09/2013-05/2014

Niedernhofer

Attended/Presented "Environmental Mutagenesis and Genomics Society 45th Annual Meeting", Orlando, FL; EMGS Forum: "Navigating a Science Career in the 21st Century"

Education Outreach 09/2013-Present

Ja Mentor Tania Rodriguez, Intern, Harriet L. Wilkes Honors College/FAU

Education Outreach 09/2014-01/2015

Kishi Mentor Luke Otfinowski, Undergraduate Intern, FAU

Education Outreach 09/2014-06/2015

Kishi Co-Mentor Jiwon Kong, Graduate Student, Seoul National University

Education Outreach 09/2014-06/2015

Niedernhofer Mentor Tommy Vo, Intern, SunCoast High School

Education Outreach 10/2013-12/2014

Ja Mentor Cathy Ray, Intern, Harriet L. Wilkes Honors College/FAU

Education Outreach 1-Jan-15 to 1-May-15

Seth Tomchik mentor to undergraduate interns Brandon Gilliland and Alisha Goldberg

Education Outreach 1-July-14 to 11-July-14

Scripps Florida Faculty, Deborah Leach-

Scampavia, Rosie Albarran-Zeckler

Kenan High School intern program

Education Outreach 1-July-14 to 15-Aug-14

Scripps Florida Faculty, Deborah Leach-

Scampavia, Rosie Albarran-Zeckler

SURF Undergraduate intern program

Education Outreach 2014-2015 Niedernhofer

Thesis Committee member, Visiting Graduate Student Jing Zhao

Education Outreach 25-Jan-15 to 1-May-15

Damon Page mentor undergrad interns Dalina Laffita and Stacy Cabral

Education Outreach 2-Apr-15 to 3-Apr-15

Baoji Xu External Graduate Student Seminars at Georgetown University, Washington, DC

Education Outreach 3-Mar-15 to 5-Mar-15

Ben Shen Seminar speaker, Boston College

Education Outreach 5/11/15 - 6/30/15 Paul Robbins Mentor Christina Bukata, graduate student

Education Outreach 6/5-30/15 Paul Robbins Mentor Heather Nick, undergraduate student Education Outreach 6/8-30/15 Paul Robbins Mentor Robert Halfon, HS Student

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Education Outreach 6/8-30/15 Paul Robbins Mentor Jordan Vo, HS Student Education Outreach 6/8-30/15 Paul Robbins Mentor Devon Wasche, SURF Education Outreach 6/9/14 -

8/15/14 Paul Robbins Mentor Colten Lankford, SURF Education Outreach 7/1/14 -

6/30/15 Paul Robbins Mentor Jing Zhao, graduate student Education Outreach 7/2014-

8/31/2014 Ja

Mentor Alina Soto Obando, Trainee, Universidad Peruana Cayetano Heredia, Peru

Education Outreach 7/30/14 - 4/22/15 Paul Robbins Mentor Priscilla Tang, graduate student

Education Outreach 8-Jun-15 to 30-Jun-2015

Scripps Faculty, Rosie Albarran-Zeckler

Kenan High School intern program

Education Outreach 8-Jun-15 to 30-Jun-2015

Scripps Faculty, Rosie Albarran-Zeckler

SURF Undergraduate intern program

Education Outreach July-14 to June-15

Chakraborty Mentor Hector Mora, intern, FAU

Education Outreach Monthly Niedernhofer

Mentor Savannah Barkdull, undergraduate, University of Virginia

Education Outreach Monthly Niedernhofer

Mentoring session for M&A Post-doctoral fellows

Education Outreach Nov-14 to Feb-15

Chakraborty Mentor to Arushi Thaper, undergraduate intern, FAU

Education Outreach Spr/Summer 2015

Katrin Karbstein Mentoring high school interns, Maria Dattolo (Jupiter, FL) and Ethan Ward (Manhattan Beach, CA)

Section 9.3(c) A schedule of the shares of stock (or other securities) held by Scripps as

payment of the royalty referred to in Section 10.2(a) and a report on any trades or activity concerning such stock (or other securities);

As noted in the Annual Report as of June 30, 2014, as partial consideration for previous license agreements with Ember Therapeutics, Xcovery and Curna, TSRI received 150,000, 263 and 107 shares, respectively, of the companies’ common stock as a “License Issue Equity” royalty. TSRI is still holding its Ember and Xcovery stock, but has liquidated its Curna stock as a result of Curna’s acquisition by Miami, FL-based Opko Health, generating proceeds of $314,000.

Section 9.4 Annual Scientific Report. Scripps shall prepare the Annual Scientific

Report that describes its scientific activities for Scripps Florida each year and deliver such annual report to Funding within one hundred twenty (120) days after the end of each fiscal year of Scripps. The form of the annual report will be substantially similar to the form Scripps uses at such time with respect to its California operations.

The Annual Scientific Report is not due until January of 2016.

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Section 9.5 Performance Expectations. Scripps, in cooperation with OTTED, shall report to Funding not less than annually on its progress in meeting certain performance expectations that reflect the aspirations of the Florida Governor and Legislature for the benefits accruing to Florida as a result of the Grant Funds. These reports shall include, but are not limited to, performance expectations addressing the following with respect to Scripps Florida;

Section 9.5(a) (Also see Section 9.5(h).) The number and dollar value of research grants

obtained by Scripps with respect to Scripps Florida from the Federal Government or sources other than Florida;

Scripps Florida scientists were awarded 71 research grants from non-Florida sources between July 1, 2014 and June 30, 2015. The total dollar amount of those grants was $51,815,211. To date, Scripps Florida scientists have been awarded research grants totaling $441,000,000.

Section 9.5(b) The percentage of total research dollars received by Scripps from sources

other than Florida, which is used to conduct research activities by Scripps in Florida;

Between July 1, 2014 and June 30, 2015, scientists at Scripps Florida expended about $57.9 million in research dollars to reimburse expenses incurred to conduct research activities at Scripps Florida. During that same time, about $11.8 million of SFFC funds were expended at Scripps Florida. Thus, about 80% of total research dollars came from sources other than Florida

Section 9.5(c) The number or value of patents obtained by Scripps with respect to

Scripps Florida;

Between June 30, 2014 and July 1, 2015, 37 foreign and domestic patent applications were filed. Since inception, 85 “families” of patent applications have been filed covering Scripps Florida technology, with each family containing 2-6 patent applications. No value has been assigned to these patents at this time.

Section 9.5(d) The number or value of licensing agreements executed by Scripps with

respect to Scripps Florida;

Two license agreements were executed between June 30, 2014 and July 1, 2015 with respect to Scripps Florida technologies.

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Section 9.5(e) The extent to which research conducted by Scripps Florida results in commercial applications;

Because of the early stage of the technology being developed at Scripps Florida and the time delay attendant to further development, no commercial therapeutic applications have emerged to date. However, as noted in previous Annual Reports, several research reagents developed at Scripps Florida continue to be available commercially through various licensing arrangements.

Section 9.5(f) The number of collaborative agreements reached and maintained with

colleges and universities in Florida and with research institutions in Florida, including agreements that foster participation in research opportunities by public and private colleges and universities and research institutions in Florida with significant minority populations, including historically black colleges and universities;

The Scripps Research Institute has developed a template entitled the Joint Cooperation Agreement (JCA) to encourage and support research collaborations with Florida institutions. Provisions are included to make it easier to collaborate on filing patents for jointly developed technologies and to share revenues from commercialized innovations. By executing these agreements in advance, we expect to streamline the scientific collaboration process between Florida organizations and Scripps Florida as they work together on biomedical research. Nine Florida institutions have currently executed this formal agreement with TSRI:

Florida International University; University of Florida; Florida Atlantic University; University of Central Florida; University of Miami; Florida State University; Nova Southeastern University; University of South Florida; Max Planck Florida Institute

Scripps scientists have also participated in formal scientific meetings with colleagues at Florida foundations, colleges and universities (See Table, below).

Science Outreach Date Participants Recipients/Event Science Outreach 1-Jul-14 Seth Tomchik Talk at FAU "In Vivo Imaging of Memory

Formation and Allocation in Drosophila." Boca Raton, FL

Science Outreach 11-Jul-14 Courtney Miller Meeting with FAU Faculty Candidate, Mohamed Kabaj

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Science Outreach 28-Jul-14 Seth Tomchik Florida Brain Initiative Meeting, Tallahassee, FL Science Outreach 4-Aug-14 Matt Disney Discussion with mayo jacksonville Science Outreach 7-Aug-14 Ron Davis Speaker: USF Health Byrd Alzheimers Institute,

Tampa FL

Science Outreach 20-Aug-14 Matt Disney Discussion with mayo jacksonville Science Outreach 09/2014 Niedernhofer Attended/Presented "The health consequences of

DNA damage", FAU Boca Raton Campus, Boca Raton, FL

Science Outreach 2-Sep-14 Matt Disney Discussion with mayo jacksonville Science Outreach 14-Sep-14 Kirill Martemyanov Max Planck Institute, Jupiter, FL Science Outreach 16-Sep-14 Roy Smith Herb Weissbach, FAU Science Outreach 22-Sep-14 Donald Phinney Drs. Elliot and Glassberg (University of Miami)

re: collaboration

Science Outreach 29-Sep-14 Laura Bohn Elucidating GPCR Functional Selectivity: Novel Opportunities for Drug Development, The New York Academy of Sciences, New York, NY

Science Outreach 10/2014 Ja Presented at "PNC Food for Thought Lunch Series", Scripps-Florida, Jupiter, FL

Science Outreach 3-Oct-14 Brock Grill Speaker: University of South Florida, Tampa Bay, FL

Science Outreach 7-Oct-14 Courtney Miller Meeting with FAU Neurscience Guest Speaker, David Dietz

Science Outreach 8-Oct-14 Hyeryun Choe Dr. Sharon Isern, Florida Gulf Coast University, Ft. Myers, FL

Science Outreach 10-Oct-14 Hyeryun Choe Dr. Sirish Namilae, Embry-Riddle Aeronautical University, Daytona Beach, FL

Science Outreach 22-Oct-14 Christoph Rader Invited Speaker, Department of Infectious Diseases, TSRI-Florida

Science Outreach 23-Oct-14 Courtney Miller Meeting with Karen Dodge, Science Outreach 24-Oct-14 Roy Smith Sancilio & Co Scientific Advisory Board

Meeting Science Outreach 11/2014 Ja Attended "Florida Biomedical Career

Symposium", Scripps-Florida, Jupiter, FL Science Outreach 18-Nov-14 Courtney Miller Karen Dodge, Science Outreach 5-Dec-14 Hyeryun Choe Reviewer, CFAR Pilot Grants, HIV-1 Vaccines

and Immunology Science Outreach 12-Dec-14 Mark Sundrud Speaker - VGTI Florida, Port St. Lucie, FL Science Outreach 13-Dec-14 Roy Smith MPFI Neuroscience Discovery Day Science Outreach 17-Dec-14 Matthew Gardner Presentation at the University of Miami Science Outreach 19-Dec-14 Niedernhofer Presented at the Biochemistry and Molecular

Biology Dept. Seminar Series at University of Miami School of Medicine, Miami, FL

Science Outreach 9-Jan-15 Brock Grill Speaker: TINNS Seminar Series, FAU Jupiter, FL

Science Outreach 23-Jan-15 Niedernhofer Invited speaker Burnett School of Biomedical Sciences, University of Central Florida

Science Outreach 30-Jan-15 Ron Davis FAU Poster Event - Synapse 2015 - Jupiter, FL Science Outreach 5-Feb-15 Thomas Kodadek Neil DeGrass Tyson seminar and lunch at the

Kravis

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Science Outreach 6-Feb-15 Mark Sundrud Speaker - FAU, Department of Basic Sciences, Boca Raton, FL

Science Outreach 27-Feb-15 Matt Disney Frenchman's Creek Luncheon Invitation Science Outreach 03/2015 Murphy Attended "MPFI Sunposium 2015: Neural

Circuits and Sunshine", Palm Beach Gardens, FL Science Outreach 04/2015 Ja Attended "Flies on the Beach Conference", FAU

Jupiter Campus, Jupiter, FL Science Outreach 9-Apr-15 Seth Tomchik Host Scripps - Florida External Seminar Series

Speaker, Shawn Xu

Science Outreach 13-Apr-15 Brock Grill Speaker: University of Miami Dept of Biology Seminar Series, Miami FL

Science Outreach 16-Apr-15 Seth Tomchik Talk LSSF Webinar Science Outreach 18-Apr-15 Damon Page, Ron

Davis, Germain Busto, Ilaria Drago, Anna

Phan, Isaac Cervantes-Sandoval, Jacob Berry, Ze Liu, Yunchao Gai,

Sophie Ziegler-Himmelreich

2015 Flies on the Beach, FAU Boca Raton Florida

Science Outreach 22-Apr-15 Matthew Gardner University of Miami Science Outreach 21-May-15 Michael Farzan Eli Gilboa, Director of the Dodson

Interdisciplinary Immunotherapy Institute, Miami, FL

Science Outreach 1-Jun-15 Damon Page Speaker: FAU CARD - Autism Breakfast, FAU Boca Raton, FL

Science Outreach 15-Jun-15 Kirill Martemyanov FASEB Meeting Science Outreach 23-Jun-15 to

25-Jun-15 Thomas Kodadek Speaker at American Peptide Symposium,

Orlando, FL

Science Outreach 27-July-14 to 28-July-14

Damon Page Speaker: Florida Brain Project Symposium, Tallahassee, FL

Science Outreach 30-Mar-15 to 31-Mar-15

Ron Davis, Germain Busto, Tugba Guven-Ozkan, Ilaria Drago,

Anna Phan, Isaac Cervantes-Sandoval, Jacob Berry, Ze Liu,

Yunchao Gai, Nathaniel Noyes, Sophie Ziegler-Himmelreich, Andrew

Giles, Scott Baker, Youjun Chen, Amy

Clipperton-Allen, Wen-Chin Huang, Julien Sejourne, Courtney

Miller, Seth Tomchik

Max Planck Florida Institute Sunposium 2015 at PGA National, Palm Beach Gardens, FL

Science Outreach 4-May-15 to 5-May-15

Andrew Giles University of South Florida - College of Marine Science: Software Carpentry, St. Petersburg, FL

Science Outreach 9-Oct-14 to 11-Oct-14

Doug Kojetin National High Magnetic Field Laboratory User Committee meeting (Tallahassee, FL)

Science Outreach Jan-Feb 2015 Niedernhofer Interviewer for the TSRI Graduate Program Science Outreach Monthly Niedernhofer Supervisory role for TSRI Histology Core

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Science Outreach ongoing Donald Phinney Life Science Technology Hub Meetings Science Outreach ongoing Christopher Haga Life Science Technology Hub Meetings Science Outreach ongoing Brock Grill Faculty Search Committee, FAU Jupiter, FL Science Outreach Sept. 13-15,

2014 Niedernhofer Attended/Presented "Environmental Mutagenesis

and Genomics Society 45th Annual Meeting", Orlando, FL

Science Outreach 28-Aug-14 Paul Robbins

Univesity of Miami Visit to meet with Dr. Camillo Ricordi

Science Outreach

28-Oct-14 Paul Robbins Meeting with Exiqon - exosomes advances, biomarker discovery and pipeline validation.

Science Outreach

19-Dec-14 Paul Robbins University of Miami - seminar - Diabetes Research Institute

Science Outreach 11-May-15 Paul Robbins To The Point Interview - Radio Show Science Outreach 28-May-15 Paul Robbins Participant - BioFlorida

Section 9.5(g) The number of collaborative partnerships established and maintained

with businesses in Florida, including small businesses;

Scripps Florida continues to maintain collaborative relationships with four Florida based companies: Opko Health, Dyadic, Vova Ida Therapeutics and Florida Power and Light.

Opko Health

Opko Health, Inc. is a publicly traded healthcare company involved in the discovery, development, and commercialization of pharmaceutical products, vaccines and diagnostic products. Opko and Scripps are currently collaborating in three major areas: the development of novel diagnostic products to detect Alzheimer’s and other diseases; the development of novel drug candidates to treat Parkinson’s Disease; and the discovery of novel antibiotics.

Dyadic

A collaborative effort between scientists at Scripps Florida and Dyadic was established to provide a complete annotation of the genome of Dyadic's proprietary fungal organism, Chrysosporium lucknowense ("C1"). The knowledge gained from this effort is expected to facilitate further development of the C1 Host Technology as a robust platform for the discovery, development and production of various materials for medical and industrial applications. Furthermore, this collaboration promotes the development of a successful biotechnology cluster in South Florida.

Dr. Richard Lerner, past President of The Scripps Research Institute, said, "In addition to its potential contributions to Dyadic's success, this partnership for our new Palm Beach County-based research group also will benefit Scripps Florida and the broader scientific community. Because relatively few fungal

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genomes have been sequenced and annotated to date, our work on Dyadic's C1 host strain will increase the body of knowledge on this important class of lower eukaryotes. We expect that the information gained through the comparative genomics of fungi will provide insights into eukaryotic cellular processes, and provide important clues for the treatment of genetic, metabolic and infectious diseases."

Vova Ida Therapeutics

Vova Ida Therapeutics is a Palm Beach County-based company founded in 2013 to commercialize research from the Corinne Lasmezas lab at Scripps Florida.

Florida Power and Light

Scripps is collaborating with Florida Power and Light, a Juno Beach, Florida-based power utility that is the principal subsidiary of NextEra Energy Inc., to develop novel and proprietary technology which may yield cheaper and more effective ways at producing fuels and other commodities from natural gas

During the past year Scripps Florida has established many partnerships with State of Florida businesses and small business entities. Below is a list of the businesses Scripps Florida has or is partnering with. 3-D MICROSCOPES INC A-1 MOVING & STORAGE

ADVANCED CASE PARTS ADVANCED PAINTING CONTRACTORS INC AERC RECYCLING SOLUTIONS AFFORDABLE DRY ICE AIR COMPRESSOR WORKS INC AIR ENERGY COMPRESSOR & VACUUM CO AIR EZE AIRCOMO AKRON BIOTECH ALL FLORIDA TOWING ALPHAGRAPHICS OF THE PALM BEACHES ANDERSON MATERIAL HANDLING ARCHIVES MANAGEMENT CENTERS INC BARNES INDUSTRIAL PLASTIC PIPING INC BEYEL BROTHERS INC BOCA BUSINESS EQUIPMENT BOCA SCIENTIFIC BRAZILIAN LIMOUSINE BREAKERS PALM BEACH BRIERS BOBCAT INC

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CAPITAL CARPET AND TILE CAPO CONSULTING C'EST SI BON CATERING CHEMPEP INC CITATION COMMUNICATIONS CLEAN FUELS OF FLORIDA CMH SOLUTIONS INC COMMERCIAL DOOR & ACCESS INC CORS-AIR COURTYARD MARRIOTT JUPITER CRYO-TECH INC CUMMINS POWER SOUTH LLC CUSTOM SIGNS TODAY DASH DOOR AND CLOSER SERVICE INC DEBON AIR MECHANICAL INC DISKOVERY EDUCATIONAL SYSTEMS CORP EASY CHAULKING & WATERPROOFING INC EMPIRE OFFICE INC FARMER & IRWIN CORP FIRE & SECURITY SOLUTIONS INC FLORIDA BEARINGS INC FLORIDA FLUID SYSTEM TECH INC FLORIDA LAMBDARAIL LLC FLORIDA PIPETTE CALIBRATIONS FLORIDA POWER & LIGHT FLOW CONTROL TECHNOLOGY GYMSOURCE GRAPHICS PLUS INC HARDRIVES OF DELRAY INC HOOVER PUMPING SYSTEMS CORPORATION INFINITY ROOFING & SHEET METAL IMPERIAL FASTENER COMPANY INC JACKSON LABORATORIES JACK WALSH CARPETS AND RUGS INC JC WHITE ARCHITECTURAL INTERIOR PRODUCTS JMB REPAIRS INC J.P. GROWERS J.R. MANNO UNIFORM AND POLICE EQUIPMENT JUPITER BEACH RESORT JUPITER ENVIRONMENTAL LABORATORIES INC JUPITER GOLF CART JUPITER MEDICAL CENTER INC JUPITER PRINTING INC KAUFMAN-DAENZER INSTRUMENTS K&M ELECTRIC SUPPLY INC KMI INTERNATIONAL INC

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KO-MAR PRODUCTIONS INC LEO A. DALY CO LILA PHOTO LJB EQUIPMENT SALES CO INC LOTSPEICH CO OF FLORIDA INC MARBLE KARE USA MAX PLANCK FLORIDA INSTITUTE MC2 INC MEDREP TECHNOLOGIES INC MICRO OPTICS OF FLORIDA INC MOLECULAR DIMENSIONS MOLLIES TROLLIES MORROW ENTERPRISES NEWTON SEATING CO NOLAN POWER GROUP LLC NOZZLE NOLEN INC OFFICE FURNITURE WAREHOUSE INC OLD FLORIDA CATERING ONE BLOOD OVER THE TOP WINDOW CLEANING INC PACE MACHINE & TOOL INC PALM BEACH HOSE & FITTINGS PENTAIR AQUATIC ECO-SYSTEMS PGA NATIONAL RESORT AND SPA PRIORITY LIMOUSINE PROSHRED SECURITY RAPID ROOTER REGAL PAINTS RIGHT WAY PLUMBING INC SHOES FOR CREWS LLC SHORELINE FLOORING SUPPLIES SIMPLE BEHAVIORAL SYSTEMS SIR SPEEDY OF TEQUESTA SMELT FEED AND PET SUPPLY INC SMITHCO SERVICES SOLID WASTE AUTHORITY SOUTH FLORIDA JANITORIAL AND POOL SUPPLY SOUTHERN WATER SERVICES INC SPEEDY ROOTER INC STAR ELECTRIC STEINER – ATLANTIC CORP SULLIVAN ELECTRIC & PUMP INC SUNBELT RENTALS SUNCHASER SYSTEMS INC SYNQUEST LABORATORIES INC TIRE KINGDOM

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TM TOOLING INC TOP CUT LAWN SERVICE INC TROPIC ENERGY SERVICES TURBOVACUUM UNIVERSAL MEDICAL SYSTEMS INC VACTEK INC VISION DATABASE SYSTEMS WEST PALM BEACH PLASTICS WILDLIFE REMOVAL SERVICES WORLD ELECTRIC WORLD PRECISION INSTRUMENT INC

Section 9.5(h) The total amount of funding received by Scripps with respect to Scripps

Florida from sources other than Funding, including a breakdown of amounts received from Grants and other sources.

Since inception through June 30, 2015, Scripps Florida has been awarded $441,000,000 in grants and sponsored research funding from state and federal agencies (including the NIH), foundations, pharmaceutical companies and other grantors. In addition, the County of Palm Beach provided $210 million to Scripps for construction of the permanent facility. Funding received by Scripps Florida from sources other than Funding for the nine months ending June 30, 2015 are set forth below:

Other Revenue sources $ 3,280,128 9 mos. ending 6/30/15 Grant Awards $ 29,197,093 9 mos. ending 6/30/15 Contributions at net present value $ 2,668,064 *9 mos. ending 6/30/15

*The amount reported above was determined in accordance with generally accepted accounting principles. Therefore, certain non-cash items, such as promises to give, are reflected at their estimated net realizable value.

Section 9.5(i) The number or value of spin off businesses created in Florida as a result of commercialization of the research of Scripps.

The four Florida companies that were created to exploit licenses to technology developed primarily at Scripps Florida, and the additional Florida company located in Jupiter to access Scripps Florida (Envoy Therapeutics), are described above. No attempt has been made by Scripps to assign a value to these spin offs, with the exception of Curna, which was purchased by Miami-based Opko Health for $10,000,000 and Envoy Therapeutics, which was purchased by Japan-based Takeda for $140,000,000.

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Section 9.5(j) The number or value of businesses that locate in Florida as a result of

Scripps Florida. Scripps cannot determine the number or value of businesses located in Florida

as a result of Scripps Florida. Section 9.5(k) The establishment and implementation of policies to promote supplier

diversity using the guidelines developed by the Office of Supplier Diversity under Section 287.09451, Florida Statutes, and to comply with the ordinances, including any small business ordinances, enacted by applicable local governments and which are applicable to Scripps Florida.

The TSRI Procurement Department, led by Mr. Adrian Orozco, Sourcing Manager/Supplier Diversity Coordinator, continues to pursue opportunities to partner with the diverse business community. Scripps Florida continues to participate in county, state and national diverse supplier shows. These shows help Scripps Florida to identify diverse businesses that can provide goods and services to the institute at a competitive price. Participation in these shows has resulted in partnerships with local companies that provide furniture, pipette calibrations, refrigeration services, relocation services, dry ice services, landscaping and irrigation services, building maintenance services, printing services, shredding services and more.

Section 9.5(l) The designation by Scripps of a representative to coordinate with the

Office of Supplier Diversity.

Mr. Adrian Orozco serves in this position as the Scripps Supplier Diversity Coordinator. Mr. Orozco represents Scripps in working with small and minority business enterprises in the State of Florida, and is actively involved in many state and local supplier diversity outreach programs. Supplier Diversity Mission and Vision Statements Mission

Our Supplier Relations and Diversity Program will integrate small and diverse businesses into the procurement process - creating awareness, ownership, and an understanding of the principals of a competitive supply base. These partnerships will maximize cost savings and efficiencies within our internal processes and our supply chain.

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Vision

We recognize the importance of a diverse supply chain and strive to develop relationships with small and diverse life science and service suppliers who can assist us in achieving our biomedical research goals. Also, we expect our strategic suppliers to establish business opportunities for small and diverse suppliers.

Section 9.5(m) The establishment and implementation of a program to conduct

workforce recruitment activities at public and private colleges and universities and community colleges in Florida, regardless of their size, which request the participation of Scripps Florida.

Scripps Florida has extended workforce recruitment efforts to Florida’s higher education institutions throughout the state. A list of Institution Career Fair and Expositions attended by Scripps Florida Human Resources Analyst, Recruiter is shown below. Event Location/Institution Date Lead/Rep Career Fair Statewide, Orlando FL 05/14/2015 Jennifer Brown

Section 4.4(c)1 Scripps shall create new jobs at Scripps Florida, the number of which

shall be measured at the end of each calendar year.

On June 30, 2015, Scripps Florida employed 585 people (521 full-time and 64 part-time).

Section 4.4(c)2 Beginning 18 months after Scripps’ occupancy of it permanent facility,

Scripps shall obtain $100,000 of non-state funding for each full-time equivalent tenured track faculty member employed at

On June 30, 2015, Scripps Florida employed 49 tenure track Faculty. By that same date, over $441 million in non-state funding had been obtained. Thus, for each tenure track Faculty, about $9 million of non-state funding had been obtained.

Section 4.4(c)3 No later than 3 years after occupancy of its permanent facility, Scripps

shall apply to the relevant accrediting agency for accreditation of its Florida graduate program.

The re-accreditation of the Scripps Ph.D. program was successfully completed in early 2011. The Doctoral Program in Chemical and Biological Sciences is a bi-coastal Ph.D. program, reflecting the “one institution/two campus” makeup of The Scripps Research Institute. Owing to the larger size and

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earlier date of establishment of the Ph.D. program on the La Jolla campus, the reaccreditation process was handled by WASC Senior College and University Commission (WSCUC). The re-accreditation process included a specific site visit and assessment of the Scripps Florida graduate program in October, 2010, by Dr. Karen Holbrook, Senior Vice President for Research, Innovation & Global Affairs, University of South Florida, and President, University of South Florida Research Foundation. As a result of the overall review and re-accreditation process, Scripps’ Graduate Program received re-accreditation for a nine-year period, effective March 7, 2011. Thus, the requirement of Section 4.4(c)3 has been satisfied, within the requirement of “no later 3 years after occupancy of its permanent facility”.

Section 4.4(c)4 Scripps shall purchase equipment for Scripps Florida [using State grant funds] according to an agreed upon schedule. Equipment purchases [acquired with State grant funds] are to be measured as of January 31st of each year.

While the equipment purchase expectations set forth in the agreed upon schedule were met several years ago, approximately $2,709,216 of equipment – acquired with State grant funds – was purchased from October 1, 2014 through June 30, 2015.

Section 4.4(c)5 Doctoral Research. No later than 18 months after occupying its

permanent facility, Scripps shall establish a program for qualified graduate students from Florida universities permitting them access to the facility for doctoral, thesis-related research.

Scripps Florida has established a Ph.D. program in 2005 ahead of the September 2010 deadline, 18 months after the anticipated occupancy of the permanent facility. Forty-nine (49) graduate students were enrolled in the Scripps Florida graduate program during the 2014-2015 academic year; three of these students defended their PhD theses and one left the program for personal reasons. A total of twenty-three students have now completed Ph.D. degrees at Scripps Florida since the establishment of the Ph.D. program in 2005. We had another successful recruiting year for new graduate students; 14 new graduate students will enter the program August 1, 2015. Efforts are made to identify and recruit highly qualified students from Florida colleges and universities to join the Scripps Florida graduate program. The Scripps Florida Graduate Admissions Committee reviews all completed applications submitted by Florida residents, or students from Florida colleges and universities, who submit applications to TSRI’s graduate program. This past year, the Scripps Florida Graduate Admissions Committee sent representatives to recruiting events at the Society for Neuroscience conference in Washington DC, national

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and regional American Chemical Society meetings, and the American Society for Biochemistry and Molecular Biology meeting. Additional recruiting efforts at national meetings are anticipated for the coming year. As the faculty ranks continue to expand at Scripps Florida over the next several years, additional efforts will be made to recruit highly qualified Florida students to the Scripps Florida Ph.D. program. Of the 54 graduate students who will be in the Scripps Florida graduate program as of August, 2015, at least seven have a Florida connection (undergraduate degrees from Florida colleges and universities, or is a native Floridian who took her/his undergraduate degree out of state). Thus, the graduate program at Scripps Florida is off to a flying start, and continues to be well ahead of schedule in meeting state requirements. In addition, Scripps Florida has entered into a Joint Education Agreement with Florida Atlantic University. In March of 2006, FAU and Scripps Florida signed a “joint education agreement” that provided a framework for planning and implementing a variety of programs to promote education and research in areas involving biomedical science and related fields. The programs envisioned include collaborations in the areas of graduate and professional education, including post-doctoral training; undergraduate education and training, including laboratory and administrative internships and, community outreach activities, including continuing education for credit and service activities. This agreement also provides a blueprint for partnerships with other educational institutions throughout the region and state to facilitate similar cooperative activities. From July 1, 2014 through June 30, 2015, forty-six FAU undergraduates performed research at Scripps Florida as research interns.

In June 2010, Scripps finalized an agreement with FAU to establish an innovative MD-PhD program. The first applications for admission into this program were received in early 2011, and two accepted the offers and matriculated into the FAU Medical School in August, 2011. One of the two students completed his undergraduate degree at University of Florida. Unfortunately, both students (in the third year of their FAU Medical School program) elected to withdraw from the joint MD-PhD program. Additional students who have received offers to join this program have declined offers and elected to enroll in MD-PhD programs at other institutions. Therefore, efforts to revamp this joint program and to enhance recruiting efforts are underway. A key issue cited by many applicants is the absence of funding (from FAU) for the MD school portion of the program; students in MD-PhD programs at other institutions receive funding from NIH Programs designed to support such programs. Thus, the FAU administration has allocated three fellowships to support MD students enrolled in the joint MD-PhD program, and efforts to seek additional funding will be pursued in the coming year.

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Section 4.4(c)6 Summer Internships. No later than 18 months after occupancy of the permanent facility, Scripps shall establish a summer internship for high school students.

Scripps Florida High School Student Summer Internship Program and Sponsored Undergraduate Summer Internship Program

Since 2005, high school students and secondary science teachers (Section 4.4(c)7) in Palm Beach County have participated in the six-week summer research internship program. In 2009, a ten-week undergraduate internship program was added to the summer program. In the summer of 2015, ten high school students, one secondary science teacher and seventeen undergraduates participated in sponsored summer internship programs. Students and teachers were placed in the Departments of: Neuroscience, Immunology and Microbial Sciences, Cancer Biology, Metabolism and Aging, and Chemistry. Support for the internship programs has been provided by the William R. Kenan, Jr. Charitable Trust, The Scripps Research Institute Graduate Studies Program and the BallenIsles Charities Foundation, Inc.

In April of 2014, a National Science Foundation (NSF) grant was awarded to Cancer Biology Associate Professor Katrin Karbstein, from the Research Experience for Undergraduates (REU) national program. Funds from the grant support Summer Undergraduate Research Fellows (SURF) on the Scripps Florida campus. Titled “SURFing the Interface between Chemistry and Biology”, the program runs concomitantly with other undergraduate programs sponsored by the Kenan Trust and TSRI. All programs are coordinated through education outreach in the office of graduate studies. The duration of the high school program continues to be six-weeks where students are placed at the “research bench” with the faculty, post-docs, and Ph.D. students working at the cutting edge of basic biomedical research. As in previous years, in 2015 the program culminated in a public presentation at the Scripps Florida campus where each student presented their research findings to Scripps researcher mentors, parents, teachers, and guest Palm Beach County students. The Kenan Fellows Facebook page continues to allow us to maintain contact and track alumni from the high school program. To date 100% of the college age alumni are pursuing or have completed post-secondary degrees at top tier universities throughout the United States. Of those who have completed their baccalaureate degree, several have continued graduate education at medical schools and doctoral graduate programs in biomedical research fields. The ten-week undergraduate program continues to elevate the intensity and independence of the research experience. Working with faculty and post-doc mentors, students are provided the research and laboratory experience needed to successfully compete in graduate school admissions and gain valuable experience outside the context of basic undergraduate laboratory instruction. The program culminates in a Scripps-wide research poster competition. This summer, the top three undergraduate posters were recognized with travel awards. They will be presenting their research at national scientific conferences during the academic year 2015-2016. As a result of the program, students return to their academic institutions able to participate in campus undergraduate poster sessions, act as ambassadors for the research and graduate programs offered at Scripps

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Florida, and enjoy an enhanced knowledge base as they continue their classroom instruction. In addition, seven alumni of sponsored Scripps Florida undergraduate internships are now pursuing doctorate degrees in The Scripps Research Institute graduate studies program.

A list of the faculty seminars for the high school and undergraduate programs can be found in Section 4.4(c)11 Seminar Series. Scripps Florida Education Outreach continues to work in collaboration with the Palm Beach County School District, to insure that all county high schools, principals, science teachers, science supervisors, and parents are aware of the annual high school program. Faculty presentations, undergraduate “ambassadors” from the high school program, and correspondence with department Chairs at targeted academic institutions provide information about the sponsored undergraduate program. Detailed descriptions and on-line applications can be found on the Scripps Florida Education Outreach web pages.

Special emphasis is placed on providing opportunities for students from populations underrepresented in the sciences (e.g. females, Hispanics, African Americans). This year, we welcomed two high school students and six undergraduate students from underrepresented groups. More importantly, when these students return to their academic institutions, they serve as mentors to other students who might come from similar backgrounds.

Eligibility / Compensation Research Internships are awarded on a competitive basis to United States citizens or permanent residents. High school students must be beginning their junior or senior year in a Palm Beach County high school in the fall preceding their summer internship. All applicants must have a minimum grade point average of 3.0 and be 16 years of age or older. Undergraduates must be a rising sophomore, junior, or senior and pursuing a post-secondary degree in a STEM subject. This year high school students were awarded a gross compensation of $9.00 per hour for the six-week summer program. Undergraduates were awarded a gross compensation of $15.13 per hour for the ten-week summer program, housing support and a weekly meal allowance of $80.00 Application Procedures The application time period for the undergraduate internship program runs year-round, however, applications submitted after February 15 are considered for the summer of the next calendar year. For the high school students, the application period is from January1st to the first Friday in March. The following is required for application to each program:

A completed application form, including the personal statement about their goals A current academic transcript; A resume/CV; Two letters of recommendation

Selection Process for 2015 Scripps Summer Intern Program

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Applications are reviewed at the close of the application period by a committee composed of education outreach staff and research faculty. A rubric is used to award points for:

completed application effective statement/essay (what is your interest in the summer internship program, what

do you hope to learn, is there a particular faculty member you wish to work with (and why))

o strength of recommendation letters (are they from STEM faculty) o STEM classes completed o GPA (from transcript)

The highest scoring applicants are matched with Scripps Florida faculty. The final selection is made by the individual faculty members.

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2015 SUMMER HIGH SCHOOL INTERNS

Name Ethnicity School

Cachae Alford Krishna Bellam Robert Halfon Jamie Harris

Nicolas Kunigk Gabrielle Marvez

Shuting (Cynthia) Mo Jordan Vo

Isaac Wendler Andrew Witmer

African Amer. Asian

Caucasian Caucasian Caucasian Hispanic

Asian Asian

Caucasian Caucasian

Palm Beach Central HS Suncoast HS

American Heritage Atlantic Community HS

Spanish River HS Seminole Ridge HS

Spanish River Community HS William T. Dwyer HS The Benjamin School The Benjamin School

2015 SUMMER UNDERGRADUATE SPONSORED INTERNS

Name Ethnicity School

Brenna Appleton Sanjay Chandrasekhar

Abigail Delawder Maria Font

Rebecca Goydel Natalie Joe

Stephanie Konecki Melia Kovach Kayla Mendez

Alice Metz Karla Montejo

Peter Rohweder Hannah Toutkoushian Megan Van der Horst Yoheilly Velazquez

Devon Wasche Nicholas Wright

White Asian White

Hispanic or Latino White

American Indian White

African Amer. Hispanic

White Hispanic

White White White

Hispanic White White

Susquehanna University University of Pennsylvania

Drury University University of Colorado

Fairmont State University Fort Lewis College

University of Illinois at Urbana ChampaignThe College of Wooster

Palm Beach Atlantic University University of Massachusetts Amherst

Florida International University The College of Wooster

Virginia Tech Hartwick College

University of Puerto Rico at Ponce Wofford College Roanoke College

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The July 2015 photograph below (taken in front of building B of the Scripps Florida research facility in Jupiter, FL) includes all of the high school interns in sponsored programs.

2015 Scripps Florida High School Interns

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The pictures below show a sampling of the 2015 Scripps Florida undergraduate summer interns.

Dr. Seth Tomchik and Dr. Tamara Boto working with undergraduate summer intern Yoheilly Velazquez

Summer Inter Peter Rohweder presenting his research poster at the end of the program

2015 Undergraduate Summer Interns from sponsored programs

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K-12 and Public Science Education Programs

The William R. Kenan, Jr. Charitable Trust, The Robert and Mary Pew Public Education Fund, The Berlin Family Foundation, and the BallenIsles Charitable Foundation supply funding for the following K-12 education programs developed through the efforts of Scripps Florida education outreach staff, faculty and research staff.

The Scripps Florida High School Career Panel An after-school interactive panel with Scripps Florida Ph.D. graduate students and post-doc fellows. Scripps scientists share experiences about their undergraduate and graduate careers and the type of research they are conducting at Scripps. The intent is to demystify the higher education/science process, encourage relationships, and answer student questions.

In the picture: Deborah Leach-Scampavia with TSRI graduate students Marie Malone, Ryan Stowe, and Spencer Wood meet with a group of high school students from Western High School. Scripps Florida Middle School Neuroscience with Kenan High School Fellows High school students from the summer intern program visit Palm Beach County middle schools. Students share their love of science, their experience as a summer researcher at Scripps and a lesson in genomics (geared for the middle school classroom). The high school students are enthusiastic role models for the younger students and are well received by the classroom teachers.

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2015 CELLebrate Science Day with Scripps Florida

Since 2009, Scripps Florida has hosted an annual public science day, sponsored by and held at The Gardens Mall in Palm Beach Gardens, Florida. Each year, scores of Scripps Florida research faculty, postdoctoral fellows, graduate students and staff interact with thousands of Palm Beach County students, parents, teachers and interested community members - all excited to learn about some of the amazing scientific breakthroughs taking place right here in Palm Beach County. CELLebrate 2015 highlighted the amazing biomedical research being done by Scripps Florida scientists with brand new displays that featured departments including Cancer Biology, Metabolism & Aging, Neuroscience, Immunology and Microbial Sciences and Molecular Therapeutics. In addition, the Chemistry and Technology groups had their own booths. The chemistry exhibit, a perennial favorite, featured everything from exploding hydrogen bubbles to the extreme cold effects of liquid nitrogen. Technology showed off Scripps Florida’s drug screening capabilities—how specialized robots help scientists search for new therapeutic compounds. Since its founding in 2004, Scripps Florida has established deep roots in the Palm Beach County community, while fulfilling its mission as a nonprofit organization to advance human health and train the next generation of scientists.

Below: thousands gather each year at The Gardens Mall to “CELLebrate Science” with Scripps Florida researchers.

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Scripps Florida Undergraduate Internships In addition to the new, sponsored summer undergraduate initiative on the Scripps Florida campus, we attempt to accommodate as many students as possible who contact us for research opportunities during the summer months. Numerous undergraduate students from Florida colleges and universities, and students from Florida who are attending college out of state, seek opportunities/incentives to return to Florida to further their research experience.

Last Name First Name Scripps FL P.I. School

ACOSTA MARY Dr. TIMOTHY SPICER NOTRE DAME

BEER JACOB Dr. PETER HODDER UNIV OF PA

HAMMOND MICHAEL Dr. KENDALL NETTLES GA COLLEGE & STATE UNIVERSITY

RAO KOUSHAL Dr. MIN GUO INDIAN RIVER STATE COLLEGE

RAVELLA REVATHI Dr. SATHYA PUTHANVEETTIL INDIAN RIVER STATE COLLEGE

REIS SANTOS KEURI GYELLI Dr. WILLIAM JA UNIVERSIDADE FEDERAL DO ABC

SALAZAR DIANA Dr. ANTHONY CARVALLOZA FLORIDA STATE UNIVERSITY

SPICER TARA Dr. PATRICK GRIFFIN UNIVERSITY OF FLORIDA

WOODY BRITTANY Dr. GAVIN RUMBAUGH UNIVERSITY OF FLORIDA

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FAU and FAU Wilkes Honors College Program In 2005 Scripps Florida established a research intern program for FAU and Wilkes Honors College students. Students receive FAU academic credit or a stipend for research performed during the school term or summer months. During the period of July 1, 2014 – June 30, 2015, thirty-four FAU undergraduate students participated in research internships at Scripps Florida. Last Name First Name Scripps FL P.I. School

BARTLETT JESSICA TIMOTHY SPICER FAU

BUKATA CHRISTINA Dr. PAUL ROBBINS FAU

CABRAL STACY Dr. DAMON PAGE FAU

CANDELA RACHAEL Dr. LAURA NIEDERNHOFER FAU

CAPI DONALD Dr. COURTNEY MILLER FAU

DAVIS DOMINIQUE Dr. MIN GUO FAU

EHRLICH MARGAUX Dr. WILLIAM JA FAU

GILLILAND BRANDON Dr. SETH TOMCHIK FAU

GIOVINAZZO VINCENZO Dr. SRINIVASA SUBRAMANIAM FAU

GREGG CALVIN Dr. KIRILL MARTEMYANOV FAU

HENNESSY-WACK DANIELLE Dr. LAURA NIEDERNHOFER FAU

KONICKI KELLIE Dr. GAVIN RUMBAUGH FAU

LAFFITA PEREZ DALINA Dr. DAMON PAGE FAU

LE PATRICIA Dr. ROY SMITH FAU

MADIRAJU SRIGITA Dr. LAURA NIEDERNHOFER FAU

MANOOGIAN KARINA Dr. BEN SHEN FAU

MORA HECTOR Dr. ANUTOSH CHAKRABORTY FAU

MURPHY ANIELLA Dr. MIN GUO FAU

NICK HEATHER Dr. PAUL ROBBINS FAU

PATEL MEERA Dr. COURTNEY MILLER FAU

PATEL SHIVANI Dr. WILLIAM JA FAU

RAJA JUNAID Dr. LAURA NIEDERNHOFER FAU

RAY CATHERINE Dr. WILLIAM JA FAU

RODRIGUEZ TANIA Dr. WILLIAM JA FAU

SABBINENI YASASVI Dr. COURTNEY MILLER FAU

SHAH NAMAN Dr. BAOJI XU FAU

SIMS CLAYTON Dr. LAURA NIEDERNHOFER FAU

SINGKORNRAT DIANA Dr. WILLIAM JA FAU

SKAF DANIEL Dr. COURTNEY MILLER FAU

TURN RACHEL Dr. PATRICIA MCDONALD FAU

VILCA SAMARA Dr. PATRICIA MCDONALD FAU

WARRAYAT REEM Dr. LAURA SOLT FAU

WEINERT-STEIN KAITLYN Dr. WILLIAM ROUSH FAU

ZIMMERMAN FRANK Dr. LAURA BOHN FAU

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Palm Beach State College (PBSC) PBSC offers two degree programs in biotechnology in response to the community need for research technicians and associates. Students enrolled in the PBSC program can receive academic credit for additional experience in the laboratory. To help students gain this experience, internships have been made available at the Scripps Florida facility as space has been available. Following is a list of students who participated as Scripps Florida interns during the July 1, 2014 – June 30, 2015 time period.

Last Name First Name Scripps FL P.I.

ALI TEVIN Dr. MATTHEW DISNEY

BRADY ANGELA Dr. SUSANA VALENTE

PEREZ YANET Dr. SRINIVASA SUBRAMANIAM Section 4.4(c)7 Research Program. No later than three years after occupancy of the

permanent facility, Scripps shall establish a research program for middle and high school teachers.

Scripps has established professional development science workshops for secondary science teachers and middle school math and science teachers. In addition, Scripps Florida offers summer internships to secondary science teachers. Scripps Florida High School Teacher Summer Internships This program allows Palm Beach County teachers to conduct basic biomedical research in a laboratory under the supervision of a Scripps Florida scientist. Teachers learn about the scientific process, research planning, experimental design, and data analysis, while interacting directly with laboratory personnel. This experience also introduces teachers to current laboratory techniques and procedures, provides information on a variety of contemporary issues in basic biomedical research, creates ties and linkages to working scientists who can assist them in curriculum development, and create opportunities for teachers to share information and knowledge with their peers. Edwin Meagher, who is a teacher at Atlantic High School in Delray Beach, worked in Dr. Brock Grill’s laboratory in the Department of Neuroscience this summer. Edwin worked on designing and implementing protocols for using C. elegans to do in class experiments on genetics, behavior and neuroscience.

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Scripps Florida Secondary and Middle School Teacher Workshops: Scripps Florida is directing greater efforts to address the needs of classroom science teachers by establishing teacher workshops in basic science, math, and laboratory skills. The “InSPIRE” programs (Instructional Support Program for Innovative Research Education) offer direct interaction with the bioscience researchers at Scripps Florida and provide greater professional

development opportunities for pre-service and in-service middle and high school science teachers in a supportive and engaging environment. Recently, the workshop was modified to fulfill needs of the teachers. Dr. Brian Paegel and his group of graduate students developed a new curriculum based on microscopy and image analysis applications in their biomedical laboratory. Portability of the lessons allows teachers to leverage the Institute’s curriculum to their own classrooms during the course of the

school year. In addition, exercises are intended to pique student curiosity for science and technology.

The program, which is supported by a grant from the National Science Foundation awarded to Dr. Paegel, provides opportunities for teachers from all of the secondary and middle schools within the Palm Beach County school district to attend the teacher workshops at Scripps Florida. Through its partnership with the school district, Scripps Florida emphasizes teacher recruitment from schools with limited resources in rural and urban Palm Beach County, particularly in areas with large underrepresented and disadvantaged student populations.

Dr. Brian Paegel working with Palm Beach County School District High School Teachers

Palm Beach County high school teachers Carolyn Slygh and Mary Fish setting up their microscopes

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Section 4.4(c)8 Adjunct Professors. No later than 18 months after occupancy of the permanent facility, Scripps shall establish a program for adjunct professors. Many current Scripps Florida Faculty have received adjunct Faculty appointments with the University of Florida, University of Miami and/or Florida Atlantic University. Such adjunct appointments are intended to provide a mechanism for graduate students enrolled in Florida research universities to collaborate with, to be co-mentored by, and to perform research in the laboratories of a Scripps Florida faculty member. A mechanism has been established for faculty members at Florida institutions who have established collaborative research programs with Scripps Florida faculty to be appointed to an Adjunct Professor position. The process is initiated by a Scripps Florida faculty member who submits a nomination to his/her department chair. If the chair concurs, the chair submits the nomination to the Office of the President for review and approval.

Current adjunct faculty:

Dr. Chris Liang of Xcovery in West Palm Beach, FL – Adjunct Associate Professor, Molecular Therapeutics Dr. Andrew Hodge of BioMotion Institute in Jupiter, FL – Adjunct Professor, Metabolism and Aging Dr. Samuel Young of Max Planck Florida Institute in Jupiter, FL – Adjunct Assistant Professor, Neuroscience Dr. Jason Christie of Max Planck Florida Institute in Jupiter, FL – Adjunct Assistant Professor, Neuroscience Dr. James Schummers of Max Planck Florida Institute in Jupiter, FL – Adjunct Assistant Professor, Neuroscience Dr. Thomas Burris of Saint Louis School of Medicine in Saint Louis, MO – Adjunct Professor, Molecular Therapeutics Dr. Stephan C. Schurer of University of Miami in Miami, FL – Adjunct Associate Professor, Molecular Therapeutics Dr. Gregg Fields of Florida Atlantic University, FL – Adjunct Professor, Chemistry

Section 4.4(c)9 Access for Science Projects. No later than 6 months after commissioning

its high throughput technology, Scripps shall establish a program to allow open access for qualified science projects.

Scripps Florida initiated the “Access to Technologies” program in January of 2006 to invite scientists from Florida universities and other academic research institutions to use state-of-the-art screening technologies at Scripps Florida’s facilities in Jupiter for qualifying projects. An additional “Core” platform is

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now available at the Scripps Florida facility that combines basic research with advanced technology. Access to Technologies Scripps Florida was created to interface cutting-edge high throughput technologies with pioneering research programs relevant to current medical needs in human diseases. One of our key goals is to develop dynamic relationships with Florida institutions to foster a knowledge-based economy that will transcend traditional barriers to moving scientific discoveries into the clinic. Florida scientists who may not have these technologies available at their respective institutions are encouraged to open the links to learn more about these Core Technologies and opportunities to access them (http://www.scripps.edu/florida/technologies/). A list of collaborative Florida researchers can be found in Section 4.4(c)10 (Collaboration with Florida Colleges and Universities). X-ray Crystallography Facility This past funding year, the core facility increased experimental capability to include services for small molecule X-ray crystallography by commissioning a Bruker AXS Smart APEX CCD diffraction system with a molybdenum sealed X-ray tube. The X-ray crystallography core facility of Scripps Florida offers state-of-the-art equipment and resources to scientists inside and outside of the Scripps FL campus by providing crystallographic analysis of their chosen biological macromolecules as well as crystalline small molecules.

The core facility offers and operates as a full service core by performing protein crystallization, X-ray diffraction data collection (both in-house and at various synchrotron sources) and processing, phasing, crystallographic refinement, model building, and visualization. The structural data obtained by the core provide scientists with a wealth of information including, but not limited to, biological functions, 3D-folding, ligand binding (small molecule or protein), or mutational effect of target macromolecules of their interests. The core also services data collection and structure determination of small molecule crystals. Crystallization Screening: For macromolecular crystallization experiments, the core facility uses commercially available 960 crystallization conditions plus various optimization reagents whose combination can make millions of different crystallization conditions. The equipment available for crystallization is: 1. A Minstrel III, an automatic plate handling and imaging system,

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2. Two RoboIncubators, which can store crystallization plates in a temperature and humidity controlled environment,

3. A Leica stereomicroscope and a high powered stereoscope for manual crystal photography, crystal screening, and crystal manipulation.

4. The TTP LabTech mosquito crystallization robot, Innovadyne 96+8 crystallization robot, and Emerald’s Matrix Maker are used to perform automated protein crystallization and optimization experiments.

Data Collection Service: Diffraction experiments are performed at an in-house x-ray facility as well as synchrotron beamlines (APS and SSRL). The equipment setup for in-house X-ray diffraction data collection consists of: 1. A Rigaku MicroMax-007 HFM X-ray generator with a VariMax HR

optics and an X-Stream 2000 crystal cryo-freezing system, 2. A Proteros Free Mounting System (FMS) for handling and

manipulating room temperature grown crystals for diffraction optimization,

3. Mar345dtb image plate detector for recording X-ray diffraction patterns,

4. Two dedicated computers in the facility for data collection and processing,

5. A Bruker AXS Smart APEX CCD diffraction system with a molybdenum sealed X-ray tube for small molecule X-ray crystallography.

Structure Determination and Analysis: Multiple data sets that were collected in-house or synchrotron are processed for macromolecular structure determination, model building, and crystallographic refinement. This past year, the core facility has produced 6 publications in major peer reviewed journals and an additional 3 manuscripts are in preparation for publication. The core facility supported 6 intramural laboratories and 2 external for their on-going grant researches. In addition, the core also supported 3 laboratories for their new grant application processes. The core facility was also actively involved in preliminary studies for grant applications of these laboratories. During this period, the core facility produced and deposited multiple macromolecular structures. Finally, the core successfully initiated a collaboration with an external laboratory that will continue until 2016.

Genomics Core The Scripps Florida Genomics Core was established to enable access by Scripps Florida and external investigators to the latest technologies for next generation sequencing and microarray analysis. These technologies allow for interrogation and subsequent comparison of the role genetics play in disease state at the global level, or at specified locations in the genome. Gene expression analysis provides a profile of

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active and inactive genes in a given tissue sample or cell type. The technologies used in the Genomics Core allow for a wide range of cost effective options for discovery on multiple platforms. Listed below are the available services provided by the Genomics Core:

* Affymetrix GeneChip Arrays:

Gene Expression Mouse 430 2.0 and Human U133 Gene ST 1.0 arrays (Human and Mouse)

* Next Generation Sequencing (Illumina NextSeq 500, SOLiD4, SOLiD5500, Ion

Personal Genome Machine, Ion Proton): RNA-Seq (mRNA, small RNA, ncRNA, profiling and discovery DNA/Seq Exome capture, Targeted resequencing and SNP analysis; analysis of nucleosome positions and chromatin modifications, pooled library screenings

* Other instruments:

Sonicators (Bioruptor and S2 Covaris) Real time PCR machines (Step One plus and Light Cycler 480) Agilent 2100 Bioanalyzer and Tapestation 2200

Florida researchers can apply for access to Scripps expertise through the Scripps “Access to Technologies” program. The Cell Based Screening Core

Researchers in the Cell-Based Screening Core leverage high-throughput technologies towards a systematic description of the function of genes encoded by the human genome, and a more comprehensive understanding of the genetic basis for human disease. The CBS group provides Scripps investigators, as well as select outside collaborators, with access to genome-wide collections of cDNAs and shRNAs that can be used to interrogate cellular models of signal transduction pathways and phenotypes.

Listed below are the available services provided by the Cell Based Screening facility:

1. TransOmic Technologies whole genome human and mouse shRNA

Libraries: PLATINUM Select MLP Retroviral shRNA libraries are next generation RNAi molecules generated using the proprietary shERWOOD algorithm developed in Dr. Gregory Hannon’s laboratory at Cold Spring Harbor Laboratory. Functional testing of 270,000 shERWOOD algorithm-based shRNA designs have allowed selection of potent shRNA designs that are highly effective even at single copy number in the human and mouse genome.

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2. Mammalian Genome Collection (MGC) cDNA Library: 16,953 human (6609 clones; 5537 unique sequences) and mouse (10,344 clones; 7718 unique sequences) cDNA sequences in an expressible vector are plated in 384 well format as 1 clone per well at a concentration of 40 ng.

3. MGC focus set generation and screening: focus sets from the MGC

cDNA library can be picked, prepped and plated in 96 or 384 well format then screened in a functional assay.

4. Gal4-Transcription Factor Library: 837 human or 721 mouse

transcription factors fused to the Gal4. DNA binding domain plated in 384 well format as 1 clone per well at a concentration of 40 ng.

5. Sigma Lopac: Library of Pharmacologically Active Compounds (1280

cmpds)

6. The Prestwick Chemical Library: Contains 1200 small molecules of 100% approved drugs by FDA, EMEA and other agencies.

The CBS core can run the screen for the investigator after the investigator has optimized the functional assay on a fee for service basis. We can also provide lipid reagents for transfection and read-out reagents such as BriteLite and AlphaScreen beads for screens. Screening Parameters: Cell line: Transfectability of cell line is paramount and requires

optimization of lipid reagent used. Controls: Positive and negative controls are highly encouraged. Plate

sets have space for 4 controls with an n=4 each (16 wells). Need minimal %CV.

Read-out: Fluorescence Luminescence Absorbance Alphascreen (homogenous bead based ELISA assay-PE) Acceptable dynamic range (signal to noise)

Florida researchers can apply for access to Scripps expertise through the Scripps “Access to Technologies” program.

Metabolic Core

The Metabolic Core offers access to validated in vitro and in vivo tests that help advance the metabolic characterization of genetic and pharmacological research models of metabolic diseases, cancer, circadian rhythms, aging and inflammation. In our laboratories we can monitor the vital signs, metabolic

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rates, body composition, feeding and sleep behavior of mice, as well as cellular and mitochondrial metabolism, hormones, nutrients and inflammatory mediators. We assist our users with trainings, experimental design and data analysis and promptly respond to eventual troubleshooting requirements during their experiments.

To accomplish our mission, we utilize the following equipment:

1. Two CLAMS units with 16 cages each (Columbus Instruments) for the real-time monitoring of whole animal metabolism. VO2 consumption, VCO2 production, locomotor activity, feeding and drinking are measured. The respiratory quotient (RER, RQ), which indicates substrate utilization (fats X carbs), is also obtained.

2. A Minispec LF-50/mq 7.5 NMR (Brucker Optics) analyzer for the non-invasive assessment of body composition of wake mice.

3. A bioDAQ system (Research Diets) for the accurate monitoring of food and liquid intake, as well as for automated pair-feeding studies.

4. Telemetry systems (Data Sciences International) for the in-vivo, real time and wireless monitoring of locomotor activity, temperature and sleep profiles.

5. A scil Vet abc (Scil) hematology analyzer yields erythrocyte, thrombocyte and total white blood cell counts, hemoglobin, and erythrocyte indices.

6. A MC4000 blood pressure and heart rate measuring system for mice (Tailcuff method, Hatteras Instruments).

7. A collar-based OxyMax Plus (Starr LifeSciences) to measure vital signs (breath rate, pulse rate, breath distension, oxygen saturation) from mice.

8. A 12 units metabolic cage system to collect urine and feces from mice (Tecniplast)

9. A Cobas c311 clinical chemistry analyzer (Roche Diagnostics) performs automated enzymatic reactions followed by the photometric reading of reaction products. There are many kits currently available. Metabolic (glucose, cholesterol, TAG, LDL, HDL), liver (transaminases, albumin, C-reactive protein, bilirubin), drugs of

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abuse (cocaine, amphetamines, alcohol and others), ammonia, creatinine are some examples.

10. A Luminex 200 (Luminex Coorporation). The Luminex allows the flow cytometric quantification of fluorescent bead-bound antibody antigen complexes. Multiplex assays for metabolic hormones, cytokines and more.

11. 24- and 96-wells Sea Horse platforms (SeaHorse Biosciences). The Seahorse instrument monitors aerobic respiration plus glycolysis in attached cells and mitochondrial preparations in real time. Additionally, it features automated delivery of substrates and compounds with activity specific to each respiratory step, enabling the characterization of mitochondrial function at the enzymatic complex level.

12. A GM7 analyzer (Analox instruments). By monitoring fluctuations in oxygen levels resulting from oxidase-based enzymatic reactions, the GM7 quantifies oxidase substrates such as glucose, glutamine, creatinine, urea, ketone bodies, glycerol, cholesterol, triglycerides, lactate, pyruvate and more in blood or plasma samples.

13. An AR-2000 radio-TLC Imaging Scanner (Eckert & Ziegler) for the detection of radiolabeled compounds in TLC plates.

The Proteomics Core The Proteomics Core at Scripps Florida offers a wide range of mass spectrometry-based proteomics services to assist with protein characterization, identification and quantification. It is essential to examine the expression and action of proteins and other gene products during normal conditions as well as disease state. The core provides support to Scripps faculty and staff who focus on such questions. In particular, the scientists concentrate on developing and applying the techniques of mass spectrometry for discovery and quantitative proteomic experiments. The core also supports the small molecule mass spectrometry needs of the institute and collaborators.

Listed below are the available services provided by the proteomics facility:

1. Proteomics: Preparation of protein samples (in-gel or in-solution proteolytic digestion) and nano-HPLC-MS/MS characterization of simple and complex protein mixtures including whole cell lysates. Mapping posttranslational modifications. Employing sample enrichment strategies for posttranslational modification analysis. Quantitative proteomics analysis of metabolic labeled (SILAC)

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samples. Characterization of chemical labeling using Tandem Mass Tags (TMT-6-plex) for quantitative proteomics analysis. Perform database searches with multiple search engines and present data with Scaffold (Proteome Software, Portland, OR).

2. Mass Spectrometry: High resolution mass spectrometry analysis of small molecules for confirmation of structure.

3. HPLC Purification: Peptides or small proteins can be purified using reverse phase separation. Detection systems include UV, multi wavelength and fluorescence.

Florida researchers can apply for access to Scripps expertise through the Scripps “Access to Technologies” program.

The Flow Cytometry Core Flow cytometry measures and analyzes the characteristics of single particles, normally cells, as they move in a stream and are passed through a laser. Thousands of cells can be analyzed by a flow cytometer in a single second. Among the measurements derived from flow cytometry are the size, relative fluorescence and complexity of the particle. Flow cytometry can be used for a variety of applications including complex cell analysis and cell sorting. The Scripps Florida Flow Cytometry Core has state of the art flow cytometry instrumentation inclusive of a BD FACS ARIA3 cell sorter, BD FACSAria Fusion cell sorter, BDLSRII analyzer, a Beckman Coulter Gallios analyzer, and a BD Canto analyzer. In addition to flow cytometry equipment, the core also offers laser capture microdissection on the Leica LMD 7000 microscope as well as hematologic analysis on the Hemavet 950FS. The BD FACSAria3 is a four-laser system configuration. The FACSAria3 uses a 488nm (blue), 561nm (yellow-green), 633nm (red), and a 407nm (violet) laser system. Sorting is available using up to sixteen fluorescence parameters as well as light scatter discrimination (cell size, organelle composition and density, and doublet discrimination). Applications include high speed, four-way cell sorting, cell purification, slide sorting, and cell cloning (by automated deposition into 6, 12, 24, 48, 96 well plates). The BD FACSAria Fusion is a four-laser system configuration. The FACSAria Fusion uses a 488nm (blue), 561nm (yellow-green), 640nm (red), and a 405nm (violet) laser system. Sorting is available using up to sixteen fluorescence parameters as well as light scatter discrimination (cell size, organelle composition and density, and doublet discrimination). Applications include high speed, four-way cell sorting, cell purification, slide sorting, and

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cell cloning (by automated deposition into 6, 12, 24, 48, 96 well plates). The FACSAria Fusion is enclosed in a ClassII Type A2 Baker biosafety cabinet consistent with BSL2 containment. The BD LSR2 is a five-laser system configuration. The LSRII uses a 488nm (blue), 561nm (yellow green), 640nm (red), 407 nm (violet), and a 355nm (UV) laser system. Analyses of up to eighteen fluorescence parameters as well as light scatter discrimination (cell size, organelle composition and density, and doublet discrimination). Applications include immuno-phenotyping, mitochondrial membrane potential, mitochondrial superoxide detection, cell proliferation, cell cycle analysis (DNA content), and calcium flux (ratiometric imaging). The Beckman Coulter Gallios is a four-laser system configuration. The Gallios uses a 488nm (blue), 561nm (yellow-green), 638nm (red), and a 405 nm (violet). Analyses of up to ten fluorescence parameters as well as light scatter discrimination (cell size, organelle composition and density, and doublet discrimination). Applications include immuno-phenotyping, mitochondrial membrane potential, mitochondrial superoxide detection, cell proliferation, and cell cycle analysis (DNA content). The Gallios features a 32-tube multi carousel loader that allows for faster tube sampling and analysis. The BD Canto is a two-laser system configuration. The Canto uses a 488nm (blue), and a 640nm (red) laser system. Analyses of up to six fluorescence parameters as well as light scatter discrimination (cell size, organelle composition and density, and doublet discrimination). Applications include immuno-phenotyping, cell proliferation and cell cycle analysis (DNA content). Other modalities (FRET, intracellular organelle staining, etc.) for analysis will be developed in response to need. The Leica LMD 7000 laser capture microdissection microscope has 4X, 6.3X, 10X, 20X, 40X, and 63x objectives, automated motorized scanning and positioning, and remote stylus operated input. Laser capture microdissection allows for the precise and contamination-free isolation of specific areas of tissue from single cells or cell groups according to morphological criteria. The microscope is also capable of cutting on any single fluorescence parameter representing commonly used fluorochromes and reporters (CFP, GFP, mCherry, etc). The dissected material is then directly accessible for further analysis. The Hemavet 950 FS is an animal blood cell counter/differential analyzer. The Hemavet allows for a fast and complete five-part WBC differential. It includes 20 parameters including platelets. The Hemavet is fully automated and simple

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to operate. Whole blood analysis is available for mouse and rat blood specimens with as little as 30ul. Tissue culture counting is also available. The Nuclear Magnetic Resonance Core Nuclear magnetic resonance, known as NMR, uses the magnetic properties of certain nuclei to identify new compounds and study molecular structure. Scripps Florida NMR core facility hosts three state-of-the-art Bruker NMR spectrometers, an Avance III 700 MHz instrument with a cryoprobe, and two AVANCE 400 MHz instruments with automation. By using those instruments, a wide variety of information can be gathered for small molecule identification and structure elucidation, protein/RNA structural and dynamic studies, and molecular interaction studies such as for compound binding screen and optimization. The three instruments run 24 hours a day, 365 days of the year. By connecting these highly sensitive instruments to the Internet via a proprietary Scripps Florida server, scientists can access the collected spectra from their office or laboratory. Florida researchers can apply for access to the NMR Core and Scripps expertise through the Scripps “Access to Technologies” program.

 

High Throughput Screening Core Description Background High Throughput Screening (HTS) is a drug-discovery process widely used in the pharmaceutical industry. It leverages automation to quickly assay the biological or biochemical activity of a large number of drug-like compounds. It is a useful for discovering ligands for receptors, enzymes, ion-channels or other pharmacological targets, or pharmacologically profiling a cellular or biochemical pathway of interest. Typically, HTS assays are performed in “automation-friendly” microtiter plates with a 384 or 1536 well format.

Capabilities The Lead Identification group at Scripps Florida has set-up a state-of-the art HTS operation to support Scripps’ intramural HTS efforts. This Core has both HTS and compound management automation, and expertise in adapting biological and biochemical bench-top assays into high-throughput screens. Listed below are the available services provided by Lead Identification Core:

1. Assay Implementation: If an assay is accepted into the Access to

Technologies Program, Scripps will use its expertise to execute an HTS assay to 384-well or 1536-well plate format as necessary.

2. Access to Technologies Compound Library: Scripps has designated

compound sub-libraries to be used for Access to Technologies

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screening efforts. These collections contain known inhibitors/activators of the most common HTS target classes (GPCRs, ion channels, etc.).

3. HTS Screening & Follow-up: Once the submitted assay has been

optimized for HTS, the "Access to Technologies" Compound Library will be screened at a single concentration. [Compounds will be assayed in triplicate to confirm activity.] Upon completion of the follow-up assay, a table of data listing the compound id#, screening concentration, and %activity (or %inhibition) for each hit compound will be provided to the researcher.

Although there are a variety of biochemical and biological assays, only a subset is amenable to HTS. The Scripps selection process includes a review of various criteria, such as assays that are amenable to automation and research programs that may have significant impact on the scientific community. Florida researchers can apply for access to Scripps expertise through the Scripps “Access to Technologies” program.

Florida HTS users:

Minond, Dmitiry Torrey Pines Institute for Molecular

Studies Liao, Daiqing University of Florida Lasmezas, Corinne The Scripps Research Institute-FL Carroll, Kate The Scripps Research Institute-FL Duckett, Derek The Scripps Research Institute-FL Kissil, Joseph The Scripps Research Institute-FL Puthanveettil, Sathyanarayanan The Scripps Research Institute-FL Rumbaugh, Gavin The Scripps Research Institute-FL Davis, Ronald The Scripps Research Institute-FL Guo, Min The Scripps Research Institute-FL Jin, Shougang University of Florida Martemyanov, Kirill The Scripps Research Institute-FL Solt, Laura The Scripps Research Institute-FL Griffin, Patrick The Scripps Research Institute-FL Kamenecka, Ted The Scripps Research Institute-FL McDonald, Patricia The Scripps Research Institute-FL

Behavior Core The Behavior Core at Scripps Florida provides state-of-the-art equipment and software for measuring rodent behavior.

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The Mouse subdivision of this Core can support most phenotyping/drug discovery test batteries as well as more focused behavioral projects. It consists of a 10-room suite that includes a dedicated procedure room (surgical and microinfusion stations), a mouse waiting room, and the following behavioral paradigms: (sensory & motor function) Grip Strength, Hot Plate, Accelerating Rota-Rod, Acoustic Startle/Pre-pulse Inhibition; (activity & emotional behavior) Open Field, Elevated Plus Maze, Tail Suspension Test, Forced Swim Test, Light-Dark Exploration; (learning, memory, & cognition) Watermaze, T-maze, Cued and Contextual Fear Conditioning, Conditioned Place Preference, Novel Object Recognition; (social behavior) Sociability and Social Novelty, and Social Interaction. The suite also has a long-term mouse holding room that contains 40 wheel running cages for Circadian Rhythm studies as well as eight home cage monitoring systems for 24/7 home cage video monitoring. The room is temperature, humidity, and light controlled and can support any required light-dark or constant dark conditions. The Rat subdivision of the Core provides equipment and software for the following behavioral paradigms: Open Field, Cued and Contextual Fear Conditioning, and Conditioned Place Preference. All behavioral rooms are fully equipped and supplied. Many of the behavioral tasks are completely automated, with software providing control over hardware and trial protocols. Standard protocols for the behavioral tasks have been developed by the Director; expertise for the development of custom protocols is available. The behavioral experiments can be fully conducted by Behavior Core staff, or individual labs can utilize the rooms and equipment for their own experiments. Training and consultation are provided free of charge. Assistance is also available for preparing the behavioral portions of IACUC protocol submissions. Behavior Core resources are also available to non-Scripps Florida scientists through collaboration with the Director. The Behavior Core officially opened for business in June of 2011. In the past year, the Behavior Core resources and/or personnel have been included on multiple Scripps Florida Faculty grant applications (many of which have been awarded funding), and data collected in the Behavior Core has been included in multiple publications. The Mouse Behavior Core has also collaborated with other institutions on research projects.

The Behavior Core added an additional staff member (research Technician) to assist with the daily upkeep and maintenance of the Behavior Core and with experiments performed by the Behavior Core.

Informatics Core

The Informatics Core provides data management and analysis services for the Scripps research community as well as for external collaborators. The

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Core offers scientific and technical support to assist in the collection, analysis, integration and dissemination of biomedical data and knowledge. Projects will often vary in time and cost depending on the scientific goals of the work, and the desired level of detail. The Core’s goal is to use existing software, tools developed by the group, along with open source software to support and advance the science of TSRI’s faculty in a cost-effective manner.

The Informatics Core has expertise analyzing data and building tools across many different scientific areas – however, the Core has identified four specific focus areas: genomics data analysis, proteomics data analysis, cheminformatics data analysis, and custom software development.

As Informatics is a data-driven field, the Informatics Core doesn’t have any physical instrumentation. However, the Informatics Core has access to and expertise in several commercial and open-source applications, including:

Ingenuity Pathway Analysis (IPA) Next Generation Sequencing tools (Tophat/Bowtie/Cufflinks, etc.) GeneSpring (Omics data analysis) R and R-Bioconductor (Data Analysis and Biostatistics ) Pipeline Pilot (Workflow Management and Automation) Schrodinger Suite (Docking, Molecular Modeling, and Computational

Chemistry) Mascot (mass spectrometry search) Sieve (label-free proteomics quantitation) HD Workbench (HD Exchange analysis)

In addition to this software, the Informatics Core has extensive experience with running applications in a high-throughput and parallel fashion on supercomputing clusters. The High-Performance Computing group, which works closely with the Informatics Core, operates two Linux clusters, Garibaldi and Sepa:

The Garibaldi Supercomputer is rated at 23.5 TFlops peak performance, and consists of:

64 Dell Poweredge M610 blades with two 2.40 GHz Intel quad core E5530 XEON-EMT processors, 48GB of ECC DDR3 memory.

96 Dell Poweredge M610 blades with two 2.27 GHz Intel quad core E5520 XEON-EMT processors, 48GB of ECC DDR3 memory.

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96 Dell Poweredge M600 blades with two 2.66 GHz Intel quad core E5430 XEON-EMT processors, 32GB of ECC DDR2 memory.

200 Dell Poweredge 1955 servers with two 2.33 GHz Intel dual core 5140 XEON-EMT processors, 8GB of DDR2 memory.

The cluster has a total of 12 TB of memory.

At the heart of the storage infrastructure is a high performance SFA10K unit from Data Direct Networks (DDN) which delivers 250 TB of distributed and persistent storage via the IBM GPFS file system. In addition each node has a 72 GB or a 146 GB local scratch space to store temporary data from computations. A large pool of object storage (using the “Ceph” open-source platform) is in the process of being brought online.

Florida researchers can apply for access to Scripps expertise through the Scripps “Access to Technologies” program.

Histology Core

The Histology Core at Scripps Florida was established in 2014 to provide full histological services as well as technical support for the investigators inside and outside of the Scripps FL campus. The Core facility is equipped with a Sakura VIP 5 tissue processor, Biocare Decloaking Chamber, Shandon rotary microtome and embedding center, routine/special stain center, a Leica CM1950 Cryostat, and a Leica BOND-MAX Automated Immunostainer.

The cryostat is available for use (at an hourly rate) by anyone who has had proper training by the Core staff. The Histology Core is capable of processing and staining investigator’s specimens for routine analysis as well as many special stains or Immunohistochemical/Immunofluorescence techniques.

Listed below are available services provided by the Histology Core:

1. Fixation Techniques

2. Tissue Processing

3. Embedding

4. Paraffin Sectioning

5. H&E’s

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6. Special Stains

7. Immunohistochemistry/Immunofluorescence

8. Cryostat Embedding and Sectioning

9. Decalcification of Bone Specimens                                                 

The Histology Core provides a broad range of special stains for paraffin or frozen sections of fixed tissues. Some of the special stains that we offer are Congo Red, Crystal Violet, Gram Stain, Luxol Fast Blue, Masson Trichrome, Oil Red O, Periodic Acid Schiff (PAS) and Safronin O. Additional stains are available upon request. The Core also offers optimization of antibodies and protocols for double and triple staining Immunohistochemistry and Immunofluorescence techniques. In addition, the Core offers consultation in staining methodology to customize the protocols in order to obtain “publication quality” results.

Florida researchers can apply for access to Scripps expertise through the Scripps “Access to Technologies” program.

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Section 4.4(c)10 Collaboration with Florida Colleges and Universities. Beginning June 2004, Scripps shall commence collaborative efforts with Florida public and private colleges and universities, and shall continue cooperative collaboration through the term of the Agreement.

On-going and new scientific collaborations between Scripps Florida scientists and colleagues from Florida colleges, universities, and local companies are described in the table below.

Scripps Florida Institutional Collaborator Collaborator(s) Institution Description of Collaboration William Roush Gregg Fields FAU Design and synthesis of inhibitors of metallomatrix proteinases William Roush Dimitriy Minond TPIMS Design and synthesis of inhibitors of

metallomatrix proteinases (MMP’s and ADAM’s)

William Roush Nagi Ayad U Miami Design, synthesis and biological characterization of inhibitors of Wee1degradation William Roush Daiqing Liao U of FL Design, synthesis and biological

characterization of inhibitors of class I HDACs and lysine acetyl transferase P300

Katrin Karbstein Elizabeth Stroupe FSU Electron microscopic analysis of pre-

ribosomal complexes and ribosome assembly factors.

Katrin Karbstein John Cleveland Moffit Cancer

Center The role of CK1delta in ribosome maturation and tumorigenesis

Sathya Puthanveettil Tom Capo U Miami Aplysia Aging Sathya Puthanveettil Lynne Fieber U Miami Aplysia Aging

Sathya Puthanveettil Leonid Moroz U of FL Aplysia Genome

Sathya Puthanveettil Robert Stackman FAU Long-term memory storage Sathya Puthanveettil Long Yan MPFI Super Resolution Microscopy

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Mark Sundrud Maria T Abreu U Miami Immunophenotypic analysis of inflammatory immune cells in Crohn’s disease patient tissues

Laura Niedernhofer William Hauswirth U of FL Aging-related loss of vision Laura Niedernhofer Janet Blanks FAU Aging-related loss of vision Laura Niedernhofer Michal Masternak UCF Role of the somatotroph axis in aging Laura Niedernhofer James Kirkland Mayo Clinic Senolytics to treat aging Laura Niedernhofer

Darren Baker Mayo Clinic

Comparison of murine models of progeria

Laura Niedernhofer Sundeep Khosla &

Joshua Farr Mayo Clinic

Bone aging

Christoph Rader Eduardo Sotomajor H. Lee Moffitt

Cancer Center & Research Institute (Tampa, FL)

Antibody-drug conjugates targeting mantle cell lymphoma

Christoph Rader Lori A. Hazlehurst H. Lee Moffitt

Cancer Center & Research Institute (Tampa, FL)

Chemically programmed antibodies targeting multiple myeloma

Christoph Rader Ronan T. Swords University of

Miami Miller School of Medicine (Miami, FL)

Biomarker discovery in acute myeloid leukemia

Paul Robbins Ghivizanni, Steve U of F Arthritis gene therapy Paul Robbins Ricordi, Camillo U of Miami Diabetes therapies, exosomes Paul Robbins Michal Masternak UCF Mouse models of aging Paul Robbins Bret Goodpaster TRI Mitochondria, aging and metabolism Paul Robbins Michal Masternak University of

Central Florida Mesenchymal stem cells from fat vs. bone marrow to treat aging

Michael Farzan Dr. Ron Desrosiers U of Miami Therapeutic effector functions of AAV-

expressed transgenes

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Dr. Deshrain Ashana

Brock Grill Ken Dawson-Scully FAU Circuit and molecular mechanisms of electroconvulsive seizure in the nematode C. elegans

Kirill Martemyanov Yuquin Li U of FL Use of genetic mouse models Kirill Martemyanov Ryohei Yasuda Max Planck

Florida Institute Imaging neuronal signaling

Kirill Martemyanov Samuel Young Max Planck

Florida Institute Electrophysiological characterization of mouse models

Kirill Martemyanov Kevin Wickmang University of

Minnesota Role of RGS proteins

Kirill Martemyanov Michelle Ehrlich Mt. Sinai Dopamine D1 Receptor/Role of RGS7 Kirill Martemyanov Laurie J. Ozelius Mt. Sinai Dopamine D1 Receptor Kirill Martemyanov Nevin Lambert Georgia Regents Matthew Disney Leonard Petrucelli Mayo Clinic,

Jacksonville, FL Inhibitors of c9RAN Translated Peptides and Toxicity in c9FTD/ALS

Thomas Bannister Claes Wahlestedt U of Miami Nociceptin Receptor Agonists for Cocaine Abuse and PTSD

Thomas Bannister John L. Cleveland Moffitt Cancer Center- Tampa

Targeting Slc16a/Mct Lactate Transporters in Cancer Therapeutics

Thomas Bannister Shouguang Jin University of Florida College of Pharmacy

Countering beta-lactam resistance in Pseudomonas aeruginosa

Susana Valente Jay McLaughlin University of

Florida dCA inhibition of Tat neurological activity

William Ja Ken Dawson-Scully FAU Drosophila aging and nutrition Shuji Kishi Matthew Gill,

William Ja, and Anutosh

Chakraborty TSRI; Kailiang Jia, FAU; and Jun-Yong Cho, Rosalind Franklin

University of Medicine and

Science

FAU Cross-species genetics of Spinster genes in development and senescence/aging

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Section 4.4(c)11 Seminar Series. Beginning 18 months after Scripps occupies the permanent facility, Scripps shall establish an annual seminar series featuring a review of the science work done by Scripps and its collaborators.

Collaborative seminars feature prominent Florida-based speakers from the academic, biotechnology or pharmaceutical communities and focus on topics within the broad fields of biomedical science, advanced technologies applied to biomedical research, drug discovery, and energy. External seminars are part of the institute series, inviting prominent researchers from national and international institutions. Both serve as a major foundation for creating knowledge- and technology-sharing opportunities, team building, and collaborations among biomedical researchers between Scripps Florida, Florida, and other research and academic institutions and companies. The sessions are open to interested professionals within the Scripps Florida and Florida scientific communities. The weekly summer intern series, an adjunct to summer intern day-to-day responsibilities, features faculty members and research associates from Scripps Florida. High school and college undergraduate interns attend specially-designed seminars throughout the course of the summer. Each seminar highlights basic science principles and the research focus/application efforts of the Scripps Florida biology, chemistry, and translational research laboratories.

External Seminars

September 18, 2014 Sidney Hecht Professor of Chemistry Arizona State University Title: Protein Synthesis with Non-alpha Amino Acids

September 25, 2014 Ryan Shenvi Assistant Professor of Chemistry The Scripps Research Institute, California Title: Chemical Synthesis of Secondary Metabolites

October 23, 2014 David Schneider Associate Professor of Microbiology and Immunology Stanford University Title: Warping disease space to improve recovery from infections

October 30, 2014 Matthew Goldberg Assistant Professor of Neurology & Neurotherapeutics, Psychiatry University of Texas, South West Medical Center Title: Analysis of Knockout Rat Models of Parkinson's Disease and LRRK2 Oligomerization

November 6, 2014 Wilfred van der Donk Professor of Chemistry University of Illinoi, Urbana-Champagne Title: Biosynthesis of Cyclic Peptide Antibiotics

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November 13, 2014 Frank Schroeder Research Group Leader, Boyce Thompson Institute Adjunct Assistant Professor, Department of Chemistry and Chemical Biology Cornell University Title: The Chemical Language of Worms: A Modular Library of Small Molecule Signals

November 20, 2014 Julien Sage Associate Professor of Pediatrics (Cancer Biology) and of Genetics Stanford University Title: Novel Therapeutic Approaches in Lung and Pancreatic Cancer

December 4, 2014 Michael VanNieuwenhze Associate Professor, Chemistry Indiana University, Bloomington Title: Novel Chemical Probes for Use in the Study of Bacterial Peptidoglycan Biosynthesis

December 11, 2014 Anna Mapp Professor of Chemistry and Director of the Program in Chemical Biology University of Michigan Title: Allosteric modulators of protein-protein interactions

January 8, 2015 Angelica Gonzalez Assistant Professor of Biomedical Engineering Yale University Title: Engineered Human Microvasculature: Composite Cellular and Matrix Structures Regulate Leukocyte Recruitment

January 15, 2015 Charles Chavkin Professor of Pharmacology University of Washington Title: Therapeutic Potential of Kappa Opioids in Pain and Addiction

January 22, 2015 Gregg Fields Professor and Chair, Dept. of Chemistry & Biochemistry Florida Atlantic University, Jupiter Adjunct Professor of Chemistry, Scripps Florida Title: Using the Mechanism of Collagenolysis to Develop Novel Matrix Metalloproteinase Probes

January 29, 2015 Paul Hanson Professor of Chemistry University of Kansas Title: Developing a Discovery Platform for Novel Electrophilic Probes: Emerging Chemotypes in Chemical Biology

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February 5, 2015 Ian Wilson Hansen Professor of Structural Biology Chair, Dept. of Integrative Structural and Computational Biology Skaggs Institute for Chemical Biology The Scripps Research Institute, California Title: Broad Neutralization of Viral Pathogens and Implications for Vaccine Design

February 12, 2015 Thomas Schwarz Professor of Neurology and Neurobiology Department of Neurology Harvard University Title: Moving and Removing Mitochondria in Axons

February 19, 2015 Jonathan Javitch Lieber Professor of Experimental Therapeutics in Psychiatry and Professor of Pharmacology (in the Center for Molecular Recognition and in Physiology & Cellular Biophysics) Columbia University Title: Single-Molecule Imaging of GPCR Organization in Living Cells

February 26, 2015 Charles Gersbach Assistant Professor of Biomedical Engineering Duke University Title: Genome and Epigenome Editing for Gene Therapy, Regenerative Medicine & Disease Modeling

March 5, 2015 Timothy Jamison Professor of Chemistry Massachusetts Institute of Technology Title: Continous Flow Multistep Synthesis

March 12, 2015 Yasmin Hurd Professor of Psychiatry/Neuroscience Mount Sinai School of Medicine Title: Cannabis, Neurodevelopment and Psychiatric Vulnerability

March 19, 2015 Eric T. Wang Principal Investigator Medical Engineering/Medical Physics, Bioinformatics & Integrative Genomics Harvard-MIT Division of Health Sciences & Technology Title: Genomic approaches to understand RNA regulation in neuromuscular disease.

April 9, 2015 X. Z. Shawn Xu Bernard W. Agranoff Collegiate Professor of the Life Sciences Life Sciences Institute, University of Michigan Professor of Molecular & Integrative

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Physiology at U-M Medical School Title: Sensory signaling in C. elegans: what can’t a worm sense?

April 16, 2015 Kent Gates Professor of Chemistry and Biochemistry University of Missouri, Columbia Title: Interstrand cross-links derived from abasic sites in duplex DNA: candidates for endogenous DNA lesions that drive aging and neurodegeneration?

April 23, 2015 Roy Parker Howard Hughes Medical Institute Professor of chemistry and biochemistry and the Cech-Leinwand Endowed Chair of Biochemistry University of Colorado Boulder Title: Assembly and properties of stress granules and P-bodies in eukaryotic cells.

April 30, 2015 George Georgiou Professor, Section of Molecular Genetics and Microbiology University of Texas, Austin Title: What’s in your Blood? System Level Analysis of Human Humoral Immunity Following Vaccination or Infection

May 7, 2015 Randy Blakely Allan D. Bass Professor of Pharmacology and Psychiatry Director Silvio O. Conte Center for Neuroscience Research & Postdoctoral Training Program in Functional Neurogenomics Vanderbilt School of Medicine Title: Synaptic Serotonin and Autism: Insights into Novel Therapies from SERT Regulatory Networks

May 8, 2015 Dale Boger Richard and Alice Cramer Professor of Chemistry Department of Chemistry Skaggs Institute for Chemical Biology Chairman, Department of Chemistry, The Scripps Research Institute Title: Redesign of Vancomycin for Resistant Bacteria

May 14, 2015 Warren Hirst Associate Research Fellow and Group Leader Neurodegeneration & Neurologic Diseases Department Pfizer Neuroscience Research Unit Title: Progress and challenges in developing novel therapeutics for the most common known causes of Parkinson’s disease: GBA and LRRK2

May 15, 2015 Joel Barrish Vice President, Head of Discovery Chemistry Bristol-Myers Squibb R&D

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Title: Innovation in Kinase Inhibitor Drug Discovery: Evolution of a Drug Target Class

May 15, 2015 Erick Carreira Professor of Organic Chemistry ETH Zürich Nobel Laureate Signature Award Title: Discovery and Surprises with Small Molecules

May 21, 2015 William Robinson Associate Professor of Medicine Immunology and Rheumatology Stanford School of Medicine Title: Sequencing Antibody Repertoires to Decipher Pathogenic Mechanisms in Rhuematoid Arthritis

Collaborative Seminars

October 16, 2014

Ann Nicole Imber, M.D., Ph.D. Florida Atlantic University “The role of Ca2+ in central respiratory control neurons of the locus coeruleus: development of chemosensitive regulation:

November 7, 2014

Ken Dawson-Scully, Ph.D. Florida Atlantic University “Invertebrate Models of Epilepsy: Uncovering Drugs and Targets for Febrile and Electronoconvulsive Seizure”

November 7, 2014

The Florida Biomedical Career Symposium Keynote Speaker: Sir Harold W. Kroto Florida State University “The Global Educational Outreach for Science, Engineering and Technology (GEOSET) Project Pioneered from Florida State University”

January 22, 2015

Gregg Fields, Ph.D. FAU, Jupiter & Scripps Florida “Using the Mechanism of Collagenolysis to Develop Novel Matrix Metalloproteinase Probes”

February 6, 2015

Dr. Samuel Young Max Planck Florida Institute “Mechanisms of Synaptic Vesicle Release Dynamics that Support the Early Stages of Auditory Processing”

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Summer Intern Seminars June 10, 2015 William R. Roush, Ph.D.

Associate Dean of the TSRI Graduate Program Topic: “Ethics in Science”

June 17, 2015 Rosie G. Albarran-Zeckler, Ph.D. Coordinator, Education Outreach Topic: “Time Management”

June 24, 2015  Matthew Pipkin, Ph.D. Associate Professor, TSRI Department of Cancer Biology Topic: “Unraveling How Chromatin Structure Regulates Cytotoxic T Lymphocyte Differentiation and Immunity”

June 9, 2015 Peter Norris and Galina Judge Environmental Health and Safety Topic: “Environmental Health & Safety Training”

June 16, 2015 Silvia Licciulli, Ph.D. Department of Cancer Biology Topic: “Trying to Fool Cancer”, The New York Times, March 2015

June 23, 2015 Erica Young, Ph.D. Department of Metabolism and Aging Topic: “Understanding Addiction”

June 30, 2015 Jenny Morgenweck, Ph.D. Department of Molecular Therapeutics Topic: “Understanding Itch”

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Section 4.4(c)12 Collaboration with OTTED. Beginning June 2004, Scripps shall commence collaboration efforts with the Office of Tourism, Trade and Economic Development by complying with reasonable requests for cooperation in economic development efforts in the biomed/biotech industry, and no later than July 2004, Scripps shall designate a person who shall be charged with assisting in these collaborative efforts.

Business outreach efforts include participation in meetings facilitated by local business and government agencies such as the Office of Tourism, Trade and Economic Development, Palm Beach County Business Development Board, and the technology Entrepreneurship & Capital Committee meeting. Similarly, community efforts involve presentations to local residential groups, various cultural organizations, and specialty groups. Numerous educational programs such as the Summer Research Internship, Science Saturday, and Introduction to Science series have been ongoing including presentation to elementary, secondary, and high schools, selecting high school students as interns, and hands-on workshops. Scientific outreach spans a variety of regional, state and international interactions from conferences, seminars and workshops; to peer-to-peer discussions. A list of such outreach programs is shown below.

Business Outreach Date Participants Recipients/Event Business Outreach 14-Jul-14 Laura Bohn, Edward

Stahl Eli Lilly Quarterly Meeting, Indianapolis, IN

Business Outreach 17-Jul-14 Ron Davis St. Mary's Medical Center - Executive Committee Meeting, West Palm Beach, FL

Business Outreach 19-Aug-14 Ron Davis St. Mary's Medical Center - Governing Board Meeting, West Palm Beach, FL

Business Outreach 27-Aug-14 Damon Page Renaissance Learning Center Board Meeting, West Palm Beach, FL

Business Outreach 16-Sep-14 Ron Davis St. Mary's Medical Center - Governing Board Meeting, West Palm Beach, FL

Business Outreach 8-Oct-14 Christoph Rader Meeting with abontek, Inc. and iBio, Inc., Seoul, Korea

Business Outreach 10-Oct-14 Christoph Rader Presentation at Celltrion, Inc, Incheon, Korea Business Outreach 10-Oct-14 Christoph Rader Presentation at Dong-A Socio Holdings, Inc.,

Yongin, Korea

Business Outreach 16-Oct-14 Chakraborty, Gill BMO Business Outreach 16-Oct-14 Paul Robbins BMO Cocktail Reception Business Outreach 21-Oct-14 Ron Davis St. Mary's Medical Center - Governing Board

Meeting, West Palm Beach, FL

Business Outreach 22-Oct-14 Niedernhofer Skype with CEO of Peter Thiel's enterprise Business Outreach 23-Oct-14 Ron Davis NAMI Advisory Council Meeting, West Palm

Beach, FL

Business Outreach 17-Nov-14 Christoph Rader Scientific Advisory Board of NBE Therapeutics in Basel, Switzerland

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Business Outreach 18-Nov-14 Christoph Rader Talk "Research Funding and Option Agreement by and between TSRI and NBE-Therapeutics: Progress by Nov 2014"

Business Outreach 18-Nov-14 Ron Davis St. Mary's Medical Center - Governing Board Meeting, West Palm Beach, FL

Business Outreach 21-Nov-14 Paul Robbins Aldabra Biosciences Business Outreach 03-Dec-14

Niedernhofer Conference call with the CEO of Alliance for Aging Research

Business Outreach 16-Dec-14 Ron Davis St. Mary's Medical Center - Governing Board Meeting, West Palm Beach, FL

Business Outreach 7-Jan-15 Thomas Kodadek Presentation at Future Energy FPL Business Outreach 14-Jan-15 Courtney Miller James Dunning and Chip Block Business Outreach 20-Jan-15 Ron Davis Mayor Muoio State of the City Address, PB

Convention Center, West Palm Beach, FL

Business Outreach 29-Jan-15 Thomas Kodadek Seminar presented at Merck, West Point, PA Business Outreach 10-Feb-15 Roy Smith PNC Event Palm Beach Business Outreach 12-Mar-15 Paul Robbins Breakfast - Paul Walczak and Joey Fago Business Outreach 17-Mar-15 Ron Davis St. Mary's Medical Center - Governing Board

Meeting, West Palm Beach, FL

Business Outreach 20-Mar-15 Niedernhofer Skype with CEO of Peter Thiel's enterprise Business Outreach 26-Mar-15 Donald Phinney Participate in BDB Life Sciences & Healthcare

Luncheon, West Palm Beach, FL

Business Outreach 27-Mar-15 Paul Robbins

Palm Beach Business Development Breakfast - Jupiter Beach Resort

Business Outreach 7-Apr-15 Paul Robbins Aldabra Biosciences - SAB teleconference Business Outreach 20-Apr-15 Damon Page Palm Beach Civic Association's Annual Meeting,

West Palm Beach, FL

Business Outreach 21-Apr-15 Roy Periana Presentation to the Board of Hyconix, Atlanta, GA

Business Outreach 21-Apr-15 Ron Davis St. Mary's Medical Center - Governing Board Meeting, West Palm Beach, FL

Business Outreach 22-Apr-15 Thomas Kodadek Meeting with Takeda Business Outreach 24-Apr-15 Niedernhofer Chamber University; NPBC Chamber Business Outreach 27-Apr-15 Ron Davis NAMI Advisory Council Meeting, West Palm

Beach, FL

Business Outreach 14-May-15 Paul Robbins Aldabra Biosciences - SAB teleconference Business Outreach 26-May-15 Ron Davis St. Mary's Medical Center - Governing Board

Meeting, West Palm Beach, FL

Business Outreach 28-May-15 Niedernhofer

Attended BioFlorida "Raising Capital for Biotech Ventures in Florida", Scripps-Florida, Jupiter, FL

Business Outreach 29-May-15 Christoph Rader Scientific Advisory Board of NBE Therapeutics in Basel, Switzerland

Business Outreach 16-Jun-15 Ron Davis St. Mary's Medical Center - Governing Board Meeting, West Palm Beach, FL

Business Outreach 17-Jun-15 Niedernhofer

Business Before Hours - State of the Chamber: Game Changer Edition

Business Outreach 16-Oct-14 to 23-Oct-14

Ben Shen ISCN28 & ICOB8, China

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Business Outreach 17-Nov-14 to 19-Nov-14

Patrick Griffin, Bruce Pascal

Thermo Fisher Meeting, Wasington, DC

Business Outreach 26-Aug-14 to 3-Sept-14

Ben Shen SIMM site visit, China

Business Outreach 27-Jul-14 to 31-Jul-14

Ben Shen ITbM 2014 site visit, Japan

Business Outreach 28-Oct-14 to 4-Nov-14

Ben Shen Visit IBC & GRC at Taipei

Business Outreach April 09-10, 2015 Niedernhofer

Host to Dr. Peter Wehling, Orthopedist, Dusseldorf, Germany

Business Outreach Monthly Niedernhofer

Board of Directors of FASEB, monthly conference call

Business Outreach Monthly Niedernhofer

Conference call with American Society for Clinical Investigation

Business Outreach Monthly Niedernhofer

Community Relations Group Scripps FL, Meetings

Business Outreach Monthly Niedernhofer

Scripps Florida representative to the Northern Palm Beach County Chamber, Trustee

Business Outreach Weekly Niedernhofer Founder and SRA with Aldabra Biosciences

*Science Outreach below is outside FL. See Section 9.5(f) for Florida Science Outreach* Science Outreach Date Participants Recipients/Event Science Outreach 1-Jul-14 Baoji Xu Gordon Conference, Hong Kong, China Science Outreach 07/2014 Niedernhofer Attended/Presented "43rd Annual Meeting of the

European Environmental Mutagen Society", Lancaster, UK

Science Outreach 07/2014 Niedernhofer Attended/Presented "The Thirteenth International Symposium on Neurobiology and Neuroendocrinology of Aging", Bregenz, Austria

Science Outreach 07/2014 Ja Attended "A Food Forum Workshop on Relationships Between the Brain, Digestive System, and Eating Behavior", Washington, DC

Science Outreach 3-Jul-14 Courtney Miller Interview with Chemistry World Magazine

Science Outreach 6-Jul-14 Paul Robbins Participant - 43rd Annual Meeting of the EEMS - Lancaster UK

Science Outreach 7-Jul-14 Ron Davis Lecturer: Okinawa Institute for Science and Technology (OIST) Conference and Workshop, Japan

Science Outreach 8-Jul-14 Tina Izard Meeting with collaborator, Dr. David Brown, University of Mississippi

Science Outreach 9-Jul-14 Courtney Miller NIH/NIDA Cutting Edge Seminar, Guest Speaker

Science Outreach 10-Jul-14 Michael Farzan KAST - Lorean Academy of Science and Technology - Los Angeles, CA

Science Outreach 10-Jul-14 Damon Page Laboratory of Molecular Biology (LMB) Alulmni Symposium, Cambridge, United Kingdom

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Science Outreach 11-Jul-14 Roy Smith Conf Call with Bill Zollers at Aratana Pharmaceuticals

Science Outreach 12-Jul-14 Matt Disney Tau Consortium meeting 9, Fort Worth, Texas

Science Outreach 13-Jul-14 Matt Disney Gordon Research Conference on posttranscriptional gene regulation

Science Outreach 13-Jul-14 Laura Bohn, Cullen Schmidt

International Narcotics Research Conference (INRC), Montreal, Quebec, Canada

Science Outreach 15-Jul-14 Paul Robbins Participant - Celegene - Warren NJ

Science Outreach 15-Jul-14 Paul Robbins Participant - Extracellular vesicles as therapeutics for autoimmune - Teleconference

Science Outreach 20-Jul-14 Ben Shen SIMB Annual Meeting, St Louis, MO

Science Outreach 24-Jul-14 Michael Farzan P01 collaborators and sub-contractors, Boston, MA

Science Outreach 27-Jul-14 Paul Robbins Participant - Neurobiology and Neuroendo – Austria

Science Outreach 28-Jul-14 Courtney Miller BYU Radio Interview (Brigham Young University)

Science Outreach 29-Jul-14 Gill Geroscience Network Meeting Report with Mayo Clinic

Science Outreach 1-Aug-14 Matt Disney American Chemical Society National Meeting

Science Outreach 08/2014 Ja Attended "2014 Colloquium on the Biology of Aging", Marine Biological Laboratory, Woods Hole, MA

Science Outreach 2-Aug-14 Paul Robbins Participant - GRC-Musculoskeletal Biology and Bioengineering - Andover NH

Science Outreach 4-Aug-14 Patrick Griffin ASMS Board Meeting, St. Louis, MO Science Outreach 8-Aug-14 Courtney Miller, Donna

Blackmond - TSRI Videoconference with Lynn Schenk, TSRI Board Member

Science Outreach 9-Aug-14 Tina Izard IUC 23rd Annual Congress Presentation - Montreal Canada

Science Outreach 11-Aug-14 Katrin Karbstein Speaker - Fassberg Semimar Series @ Max Planck Institute for Biophysical Chemistry, Germany

Science Outreach 13-Aug-14 Michael Farzan Presentation at University of Florida - Gainsville, FL

Science Outreach 13-Aug-14 Tina Izard Meeting with collaborator, Dr. Sygush with University of Montreal - Montreal, Canada

Science Outreach 14-Aug-14 Roy Smith Meeting with Fei Xu, iHuman Institute of Shanghai Tech University

Science Outreach 15-Aug-14 Christoph Rader Presentation at Scripps, La Jolla - Carlos F. Barbas III Memorial Symposium

Science Outreach 17-Aug-14 Mark Sundrud Speaker - Division of Immunobiology & The Center for Systems Immunology, Cincinnati, OH, USA

Science Outreach 23-Aug-14 Laura Bohn Pratical Information Workshop: Exploring the Biology of GPCRs, Leiden Amsterdam, Netherlands

Science Outreach 24-Aug-14 Doug Kojetin ICMRBS XXVI Science Outreach 25-Aug-14 Matt Disney Accessible Media, Inc talk about science in

Disney lab Science Outreach 26-Aug-14 Roy Smith Kimberly Cameron and Marie-Laure Rives from

Pfizer

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Science Outreach 28-Aug-14 Doug Kojetin Seminar, NC State University, Department of Biochemistry

Science Outreach 1-Sep-14 Kirill Martemyanov Max Planck Institute, Jupiter, FL Science Outreach 1-Sep-14 Kirill Martemyanov National Physics Laboratory, England Science Outreach 1-Sep-14 Kirill Martemyanov University Pierre et Marie Curie, CNRS, France Science Outreach 09/2014 Niedernhofer Attended "26th Annual Fanconi Anemia

Research Fund Scientific Symposium", Bethesda, MD

Science Outreach 09/2014 Niedernhofer Attended/Session Chair "Cold Spring Harbor Laboratory 2014 Meeting - Molecular Genetics of Aging", Cold Spring Harbor, NY

Science Outreach 09/2014 Ja Attended/Presented "Food Matters in Drosophila Lifespan", University of Alabama at Birmingham; Birmingham, AL

Science Outreach 5-Sep-14 Roy Smith Baylor College of Medicine Science Outreach 8-Sep-14 Courtney Miller NIH/NIDA Cocaine Targets Meeting, Invited

Guest Participant

Science Outreach 8-Sep-14 Patrick Griffin Invited speaker, DiaSorin, Inc., Stillwater, Minnesota

Science Outreach 9-Sep-14 Courtney Miller Meeting with Rebecca Farkas, NIH/NINDS Science Outreach 9-Sep-14 Paul Robbins Participant - Frontiers in Regenerative Medicine

Meeting - Teleconference

Science Outreach 12-Sep-14 Matt Disney Myotonic Dystrophy conference Science Outreach 13-Sep-14 Paul Robbins Participant - EMGS Annual Meeting - Orlando

FL Science Outreach 14-Sep-14 Gavin Rumbaugh Biomedica San Pau, Spain Seminar Presentation Science Outreach 14-Sep-14 Patrick Griffin JMB Editorial Board Meeting, San Francisco, CA Science Outreach 15-Sep-14 Gavin Rumbaugh University of Edinburgh, Scotland Seminar

Presentation

Science Outreach 15-Sep-14 Thomas Kodadek DARPA Kick Off Meeting-Telluride, CO Science Outreach 16-Sep-14 Brock Grill Cold Spring Harbor: Axon Guidance, Synapse

Formation and Regeneration.

Science Outreach 16-Sep-14 Brock Grill NSF Neural Systems Cluster study section, Arlington VA

Science Outreach 19-Sep-14 Roy Smtih Conf Call with Claudio Pietra, Helsinn Therapeutics

Science Outreach 19-Sep-14 Roy Smith Conf Call with Pfizer Science Outreach 20-Sep-14 Patrick Griffin Invited speaker, SIMM/CAS, Shanghai, China Science Outreach 21-Sep-14 Ron Davis Janelia Farms: A Synthesis of Bees and Flies,

Virginia

Science Outreach 22-Sep-14 Seth Tomchik Janelia Flies and Honeybee Learning and Memory, Washington, DC

Science Outreach 28-Sep-14 Paul Robbins Seminar - Molecular Genetics of Aging - Cold Springs Harbor NY

Science Outreach 29-Sept-14 Gill Attend Cold Spring Harbor Laboratory Meeting, NY

Science Outreach 1-Oct-14 Doug Kojetin Seminar, Emory University, Department of Biochemistry

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Science Outreach 10/2014 Niedernhofer Attended "The 5th Annual Mayo Clinic Robert and Arlene Kogod Center on Aging Conference", Rochester, MN

Science Outreach 10/2014 Ja Attended "Annual Student Symposium", Lake Arrowhead, CA

Science Outreach 2-Oct-14 Courtney Miller NIH/BRLE Center for Scientific Review Meeting, Invited Guest Participant

Science Outreach 3-Oct-14 Roy Smith Conf call with Bill Zollers, Aratana Pharmaceuticals

Science Outreach 10/3/2014 Chakraborty EGG Grant review and decision Science Outreach 4-Oct-14 Ron Davis and E. Nick

Petersen Neurofly 2014 - European Fly Neurobiology, Conference, Crete Greece

Science Outreach 6-Oct-14 Paul Robbins Participant - Korean Society of Gene and Cell Therapy - Seoul Korea

Science Outreach 7-Oct-14 Christoph Rader Invited Speaker at 2014 Osong BioExcellence Conference at Osong New Drug Development Center in Osong, Korea

Science Outreach 8-Oct-14 Christoph Rader Invited Speaker at Seoul National University, Seoul, Korea

Science Outreach 12-Oct-14 Matt Disney ALS TDI Leadership Summit Discussion Science Outreach 14-Oct-14 Michael Farzan 2014 Strategies for an HIV Cure at NIAID,

Bethesda, MD

Science Outreach 14-Oct-14 Kirill Martemyanov External Seminar Series, TSRI, La Jolla Science Outreach 14-Oct-14 Kendall Nettles Invited Speaker: Predicting Phenotypes for

Estrogen Receptor Ligands, UT Southwestern Medical Center at Dallas, TX

Science Outreach 16-Oct-14 Patrick Griffin DMP Study Section, New Orleans, LA Science Outreach 16-Oct-14 Laura Bohn Functional Selectivity at NIDA, Seminar,

Washington, DC

Science Outreach 20-Oct-14 Katrin Karbstein Reviewer - NSF Biology REU Panel 150222, Arlington, VA, USA

Science Outreach 21-Oct-14 Doug Kojetin AHA Lipids Basic Science # Committee Spring 2015

Science Outreach 23-Oct-14 Matt Disney Keynote speaker SUNY Albany RNA Center Science Outreach 23-Oct-14 Paul Robbins Participant - Mayo-Groningen - Rochester MN Science Outreach 24-Oct-14 Ben Shen The 8th Sino - US CBDD, Changsha Science Outreach 25-Oct-14 Roy Periana University of Tokyo, and Keio University, Murai

Symposium

Science Outreach 27-Oct-14 Kate Carroll Mike Radtke, NIH SBCA Review Meeting, New Orleans, LA

Science Outreach 28-Oct-14 Brock Grill NIH/NDPR study section, Arlington VA Science Outreach 29-Oct-14 Christoph Rader Invited Speaker at TSRI La Jolla, California on

10/29/14

Science Outreach 30-Oct-14 Hyeryun Choe Dr. Jonathan Abraham, Harvard Medical School, Boston, MA

Science Outreach 30-Oct-14 Katrin Karbstein Speaker - M-LSA Chemistry, University of Michigan, Ann Arbor, MI, USA

Science Outreach 30-Oct-14 Patrick Griffin CDRD Board of Directors Meeting, Vancouver, BC, Canada

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Science Outreach 31-Oct-14 Courtney Miller NPR Interview Science Outreach 11/2014 Niedernhofer Attended/Presented "R24 Geroscience Network

Coordinating Group Meeting and GSA Meeting", Washington, DC

Science Outreach 11/2014 Niedernhofer Attended/Presented/Meeting Co-organizer "5th US-EU Conference on Repair of Endogenous DNA Damage", Santa Fe, NM

Science Outreach 11/2014 Niedernhofer Attended/Presented at "The Xeroderma Pigmentosum Family Support Group", Kansas City, MO

Science Outreach 4-Nov-14 Roy Smith Prader Willi Syndrome Annual Meeting, Invited Guest

Science Outreach 4-Nov-14 Chakraborty Attend Obesity Week, Boston, MA Science Outreach 5-Nov-14 Jun-Li Luo NIH Review Board Science Outreach 5-Nov-14 Paul Robbins Participant - R24 Geroscience Network

Coordinating Group Meeting - Washington DC

Science Outreach 6-Nov-14 Ron Davis Molecular Psychiatry Association - San Francisco Conference, California

Science Outreach 6-Nov-14 Paul Robbins Participant - Xeroderma Pigmentosum Family Support Group - Kansas City MO

Science Outreach 7-Nov-14 Matt Disney ALS TDI Leadership Summit Talk Science Outreach 9-Nov-14 Christoph Rader Invited Speaker at 2014 Cold Spring Harbor

Engineering & Phage Display Course on 11/9/14 in Cold Spring Harbor, New York

Science Outreach 10-Nov-14 Laura Bohn Anesthesiology Research Seminar, St. Louis, MO Science Outreach 11-Nov-14 Paul Robbins Participant - US EU DNA Repair Meeting - Sante

Fe NM

Science Outreach 14-Nov-14 Courtney Miller NIH/NIDA Frontiers in Addiction Research Mini Convention, Invited Guest Participant

Science Outreach 14-Nov-14 Kirill Martemyanov Mt. Sinai Medical School, New York Science Outreach 14-Nov-14 Matt Disney C9 orf72 discussion with Target ALS group Science Outreach 14-Nov-14 Ron Davis, Jacob Berry,

Germain Busto, Yunchao Gai, Ze Liu, E. Nick Petersen, Courtney MacMullen, Courtney Miller, Seth Tomchik

SFN Annual Meeting, Washington DC

Science Outreach 15-Nov-14 Gavin Rumbaugh NIH Study Section Science Outreach 15-Nov-14 Gavin Rumbaugh SFN Meeting Science Outreach 17-Nov-14 Paul Robbins Participant - Baxter Meeting - Chicago IL Science Outreach 19-Nov-14 Christoph Rader Meeting with Institute of Biochemistry,

University of Zurich in Zurich, Switzerland re: collaboration

Science Outreach 19-Nov-14 Damon Page Rhett Syndrome Review Board, Chicago, ILL Science Outreach 20-Nov-14 Christoph Rader Meeting with Department of Medicine II-

Hem/Onc, University of Würzburg in Würzburg, Germany re: collaboration

Science Outreach 20-Nov-14 Thomas Kodadek Lecture at UTSW Dept. of Pharmacology Seminar Series, Dallas, TX

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Science Outreach 21-Nov-14 Thomas Kodadek Lecture at UTD Chemistry Seminar Series, Dallas, TX

Science Outreach 22-Nov-14 Tina Izard Meeting with collaborator, Dr. Michael Rossman with Perdue University - West Lafayette, Indiana

Science Outreach 12/2014 Niedernhofer Attended FASEB Board of Directors Meeting, Arlington, VA

Science Outreach 7-Dec-14 Courtney Miller ACNP Annual Meeting (American College of Neuropsychopharmacology)

Science Outreach 8-Dec-14 Seth Tomchik Vanderbilt University, Nashville, TN “Mechanisms of memory encoding in Drosophila”

Science Outreach 9-Dec-14 Roy Smith The Conference on Bioactive Peptides for Cell-Cell Communication, Invited Guest, Kyoto, Japan

Science Outreach 10-Dec-14 Matt Disney Discussion with NIH Science Outreach 11-Dec-14 Christoph Rader Invited Speaker - Professor Shabat Graduate

Class, Tel Aviv University, Tel Aviv, Israel

Science Outreach 14-Dec-14 Christoph Rader Invited Speaker - TAU Research Group, Tel Aviv University, Tel Aviv, Israel

Science Outreach 15-Dec-14 Christoph Rader Meeting with Dr. Ronit Satchi-Fainaro, TAU Research Group, Tel Aviv University, Tel Aviv, Israel re: Collaboration and previous days talk

Science Outreach 15-Dec-14 Matt Disney Talk Brainstorming DM Science Outreach 15-Dec-14 Scott Hansen NIH Common Fund High Risk-High Reward

Research Symposium (Bethesda, MD)

Science Outreach 17-Dec-14 Roy Smith Conf Call with Bill Zollers at Aratana Pharmaceuticals

Science Outreach 22-Dec-14 Matt Disney Visit from Patrick Brannely from the TAU Consortium

Science Outreach 11-Jan-15 Matthew Gardner Presentation and Talk at Harvard Medical School, Boston, MA

Science Outreach 11-Jan-15 Ben Shen NP Discovery & Development, SIMB, San Diego, CA

Science Outreach 13-Jan-15 Thomas Kodadek TSRI Faculty Lecture, La Jolla, CA Science Outreach 14-Jan-15 Patrick Griffin Invited speaker CFI (Canada Foundation for

Innovation) Multidisciplinary Assessment Committee, Ottawa, Canada

Science Outreach 19-Jan-15 Courtney Miller Sunovion Pharmaceuticals, Invited Guest Speaker Science Outreach 21-Jan-15 Matt Disney Tau Consortium meeting 10, San Francisco, CA Science Outreach 27-Jan-15 Katrin Karbstein Attended Peer Review Committee for RNA

Mechanisms in Cancer, ACS, Atlanta, GA, USA

Science Outreach 31-Jan-15 Laura Bohn Gordon Research Conferences, Ventura, CA Science Outreach 1-Feb-15 Kirill Martemyanov University of Texas, Houston Science Outreach 7-Feb-15 Doug Kojetin Biophysical Society 59th Annual Meeting Science Outreach 8-Feb-15 Kate Carroll Mike Radtke, NIH SBCA Review Meeting,

Washington, DC

Science Outreach 8-Feb-15 Paul Robbins Seminar - GRC Mammalian DNA Repair - Ventura CA

Science Outreach 10-Feb-15 Courtney Miller Interview with Ozy.com Science Outreach 12-Feb-15 Matt Disney Seminar Department of Cehmistry University of

Alabama Birmingham

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Science Outreach 12-Feb-15 Doug Kojetin NIH MSFB study section Science Outreach 12-Feb-15 Patrick Griffin DMP Study Section, Washington, DC Science Outreach 15-Feb-15 Gavin Rumbaugh UCI Seminar Presentation Science Outreach 15-Feb-15 Ben Shen Frontiers in Biomedical Research Symposium,

Indian Wells, CA

Science Outreach 15-Feb-15 Ron Davis 2015 Frontiers in Biomedical Research Symposium, Indian Wells, CA

Science Outreach 15-Feb-15 Roy Smith Frontiers Biomecial Research Symposium Science Outreach 15-Feb-15 Laura Bohn 2015 Frontiers in Biomedical Research

Symposium, Indian Wells, CA

Science Outreach 17-Feb-15 Matt Disney Seminar Novartis in San Francisco, CA Science Outreach 18-Feb-15 Matthew Gardner Present at AIDS Vacinne Research Lab in

Madison, WI

Science Outreach 18-Feb-15 Matt Disney Seminary Chemistry and Chemical Engineering and Biochemistry Stanford University

Science Outreach 18-Feb-15 Courtney Miller Neurotherapeutics Discovery and Development Course, Invited Participant

Science Outreach 18-Feb-15 Paul Robbins Participant - Exosome Meeting - Italy Science Outreach 19-Feb-15 Baoji Xu NIH Neurological Sciences and Disorders B

study section

Science Outreach 21-Feb-15 Michael Farzan Present at Conference on Retrovirology and Opportunistic Infections (CROI) in Seattle, WA

Science Outreach 22-Feb-15 Laura Bohn NIH Grant R01 DA031927, University of Kansas KAPPA Meeting, Jupiter, FL

Science Outreach 26-Feb-15 Patrick Griffin Invited speaker at University of Missouri (Host Stefanos Sarafinos, PhD)

Science Outreach 03/2015 Niedernhofer Host Dr. Janine Kruit (expert in endocrinology and metabolism), Assistant Professor, University Medical Center of Groningen, to help with experiments

Science Outreach 03/2015 Ja, Deshpande Attended "56th Annual Drosophila Research Conference", Chicago, IL

Science Outreach 2-Mar-15 Matt Disney AstraZeneca Seminar Cambridge, MA Science Outreach 3-Mar-15 Mark Sundrud Reviewer - NIH/NIAID Review Special

Emphasis Panel, Bethesda, MD, USA

Science Outreach 4-Mar-15 Matthew Gardner Present at Palm Springs Symposium on HIV/AIDS in Palm Springs, CA

Science Outreach 4-Mar-15 Baoji Xu Seminar presenter at University of Alabama at Birmingham

Science Outreach 8-Mar-15 Paul Robbins Participant - MIRM Retreat - Nemacolin Woodlands

Science Outreach 9-Mar-15 Hyeryun Choe Reviewer, ISF (The Israel Science Foundation) Research Grants

Science Outreach 10-Mar-15 Ron Davis NIH/NINDS Review Panel Meeting, Arlington VA

Science Outreach 12-Mar-15 Thomas Kodadek Lecture at Sigma-Aldrich Science Outreach 12-Mar-15 Matt Disney Myotonic Dystrophy Foundation Science

Advising Science Outreach 13-Mar-15 Kimberly Lovell Behavior, Biology and Chemistry: Translational

Research in Addition, San Antonio, TX

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Science Outreach 15-Mar-15 Kirill Martemyanov ASPET Conference Science Outreach 15-Mar-15 Gavin Rumbaugh NIH Study Section Science Outreach 15-Mar-15 Patrick Griffin Invited speaker - Univeristy of Phoenix, Phoenix,

Arizona

Science Outreach 20-Mar-15 Matt Disney Seminar at Johns Hopkins Medical Center Science Outreach 21-Mar-15 Michael Farzan Presentation at Keystone Symposia in Banff,

Canada Science Outreach 21-Mar-15 Laura Bohn American Chemical Society, Division of

Medicinal Chemistry, Sumposium, Denver, CO

Science Outreach 22-Mar-15 Matt Disney American Chemical Society Meeting Denver, CO Science Outreach 23-Mar-15 Jun-Li Luo NIH Review Board Science Outreach 23-Mar-15 Patrick Griffin Invited speaker at Keystone Symposia, Whistler,

British Columbia, Canada

Science Outreach 24-Mar-15 Wen-Ching Huang Wiring the Brain Meeting at Cold Spring Harbor, New York

Science Outreach 24-Mar-15 Thomas Kodadek DARPA Review Meeting, Arlington, VA Science Outreach 25-Mar-15 Matt Disney Discussion with Rutgers University regarding

collaborative project

Science Outreach 26-Mar-15 Matt Disney Discussion with NIH Science Outreach 27-Mar-15 Laura Bohn, Edward

Stahl Experimental Biology 2015 Annual Meeting, Boston, MA

Science Outreach 28-Mar-15 Brock Grill Co-Chair: ASPET Annual Meeting in Boston, MA

Science Outreach 29-Mar-15 Tina Izard 2015 ASBMB Annual Meeting Science Outreach 29-Mar-15 Patrick Griffin Invited speaker at ASPET (American Society for

Pharmacology and Experimental Therapeutics) Symposium 2015, Boston, MA

Science Outreach 29-Mar-15 Mi Ra Chang Attended ASPET Conference - presented poster, Boston, MA

Science Outreach 30-Mar-15 Patrick Griffin Molecular Pharmacology Editorial Board meeting

Science Outreach 30-Mar-15 Tina Izard Meeting with collaborator, Dr. Robert Stroud, University of California, San Franscisco

Science Outreach 31-Mar-15 Matt Disney Margaux's Miracle Foundation about Childhood Ewing's Sarcoma Research

Science Outreach 1-Apr-15 Courtney Miller NIH/NIDA Special Emphasis Review Panel Science Outreach 2-Apr-15 Katrin Karbstein Invited Speaker & Editorial Board member -

2015 ASBMB Annual Meeting, Boston, MA, USA

Science Outreach 9-Apr-15 Patrick Griffin Invited speaker - University of Texas, Arlington, Texas

Science Outreach 13-Apr-15 Baoji Xu Keystone Symposium, Snowbird, Utah Science Outreach 13-Apr-15 Matt Disney Seminar Chemistry and Biosciences Departments

University of Chicago

Science Outreach 14-Apr-15 Matt Disney Target ALS Meeting NYC Science Outreach 15-Apr-15 Sathya Puthanveettil Columbia University Presentation, New York Science Outreach 20-Apr-15 Jenny Morgenweck Kappa Therapeutic Conference 2015 Third

Conference on the Therapeutic Potential of Kappa Opioids in Pain and Additiona, Chapel Hill, CA

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Science Outreach 23-Apr-15 Thomas Kodadek NIH Director’s New Innovator Award Review meeting

Science Outreach 23-Apr-15 Patrick Griffin CDRD Board of Directors Meeting, Vancouver, BC, Canada

Science Outreach 24-Apr-15 Corinne Lasmezas Presentation for the Scripps Florida Luncheon series "Food for thoughts"

Science Outreach 24-Apr-15 Courtney Miller Society on NeuroImmune Pharmacology (SNIP), Invited Participant

Science Outreach 30-Apr-15 Roy Smith Conf Call with Claudio Pietra, Helsinn Therapeutics

Science Outreach 05/2015 Niedernhofer Attended/Presented/organizer "Geropathology Research Network Symposium", University of Washington South Lake Union Campus, Seattle, WA

Science Outreach 05/2015 Niedernhofer Attended "Frontiers in Aging and Regeneration Research", Marine Biological Laboratory, Woods Hole, MA

Science Outreach 4-May-15 Kate Carroll Drs. Derek Tan and Alex Kentsis, Memorial Sloan Kettering Cancer Center, New York

Science Outreach 5-May-15 Kate Carroll Drs. Tom Muir and Dr. Van Zandt Williams, Princeton University, Princeton, NJ

Science Outreach 6-May-15 Doug Kojetin AHA Greater Southeast Affiliate Research Committee

Science Outreach 6-May-15 Paul Robbins Participant - R24 Pathology of Aging Network Symposium - Seattle

Education Outreach 8-May-15 Howard Petrie Attend American Association of Immunologists "Immunology 2015" Annual Meeting in New Orleans, LA

Science Outreach 8-May-15 Matt Disney NIH Pioneer Appicant Stage 2 Review Science Outreach 13-May-15 Michael Farzan Guest speaker - Merck Cure Roundtable,

Philadelphia, PA

Science Outreach 13-May-15 Damon Page, Amy Clipperton Allen,

Youjun Chen, Wen-Chin Huang

International Meeting for Autism Research in Salt Lake City, UT

Science Outreach 15-May-15 Sathya Puthanveettil Washington State University Seminar Presentation

Science Outreach 16-May-15 Laura Bohn ASCEPT-BPS Joint Scientific Meeting, Hong Kong

Science Outreach 19-May-15 Corinne Lasmezas Continuous Medical Education Presentation at VA Medical Center, West Palm Beach, FL

Science Outreach 26-May-15 Corinne Lasmezas Prion 2015 Working Conference, Ft. Collins, CO Science Outreach 26-May-15 Katrin Karbstein Attended 20th Annual Meeting of RNA Society,

Madison, WI, USA

Science Outreach 26-May-15 Donald Phinney ISCT 2015 Annual Meeting, Las Vegas, Nevada Science Outreach 29-May-15 Roy Smith iHuman Institute at Shanghai Tech, Invited Guest

Speaker

Science Outreach 30-May-15 Matt Disney Tau Consortium Deep Dive, Cambridge MA Science Outreach 31-May-15 Ben Shen ASM Meeting, New Orleans

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Science Outreach 31-May-15 Devrishi Goswami, Venkat Dharmarajan

ASMS Meeting - presented poster, St. Louis, MO

Science Outreach 31-May-15 Patrick Griffin ASMS Board of Directors Meeting, St. Louis, MO

Science Outreach 1-Jun-15 Kendall Nettles 2015 ASCO Annual Meeting, IL Science Outreach 06/2015 Ja Attended "3rd Biennial Conference of the North

American Society for Comparative Endocrinology", Ottawa, Ontario, Canada

Science Outreach 2-Jun-15 Michael Farzan Present at NIAID Development of Monoclonal Antibodies for HIV in Rockville, MD

Science Outreach 2-Jun-15 Roy Periana ADHOC2015 International Symposium Speaker, Madison, WI

Science Outreach 3-Jun-15 Laura Solt NIH Study Section Participant Science Outreach 4-Jun-15 Paul Robbins Participant - Mayo - C-Sig Minisymposium -

Rochester MN

Science Outreach 10-Jun-15 Kate Carroll Mike Radtke, NIH SBCA Review Meeting, Seattle, WA

Science Outreach 10-Jun-15 Doug Kojetin NIH MSFB study section Science Outreach 14-Jun-15 Matt Disney Gordon Research Conference Invited Speaker

High throughput screening and chemistry

Science Outreach 15-Jun-15 Gavin Rumbaugh Korean KAVLI Conference Science Outreach 19-Jun-15 Michael Farzan Present at Gordon Research Conference - Viruses

& Cells in Girona, Spain

Science Outreach 24-Jun-15 Donald Phinney Participant in Science Webinar - Sourcing Niche Cell Populations: Techniques for Isolating and Characterizing Progenitor Cells

Science Outreach 24-Jun-15 Baoji Xu 15th International Symposium by the Society of Chinese Bioscientists in America, Taipei, Taiwan

Science Outreach 24-Jun-15 Patrick Griffin CDRD Board of Directors Meeting, Vancouver, BC, Canada

Science Outreach 29-Jun-15 Mark Sundrud Speaker - Pathology Seminar Series, NYU School of Medicine, New York, NY, USA

Science Outreach 30-Jun-15 Matt Disney Gordon Research Conference Invited Speaker Nucleotides and Nucleosides

Science Outreach 21-Jul-15 Tina Izard Meeting with collaborator, Dr. Gerard Bricogne, with Global Phasing Limited

Science Outreach 12/30/2014-01/03/2015

Niedernhofer Hosted to Dr. Robert Sobol, University of Pittsburgh

Science Outreach 12-Nov-14 to 14-Nov-14

Ben Shen Iowa State University, Seminar

Science Outreach 16-Nov-2014 to

19-Nov-2014

Mark Sundrud Speaker - Concordia/NDSU, Moorshead, MN

Science Outreach 2014 2015

Chakraborty The Obesity Society Scientific Review Committee Conference Call

Science Outreach 21-Mar-15 to 24-Mar-15

Roy Periana, Michael Konnick, Brian

Hashiguchi

249th ACS National Meeting & Exposition, Denver, CO

Science Outreach 28-Jun-15 to 3-July-15

Scott Hansen FASEB - Ion Channel Regulation

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Science Outreach 9-Mar-15 to 10-Mar-15

Michael Farzan Meet with Gates Foundation, Seattle, WA

Science Outreach August 04-05, 2014

Niedernhofer NIH study section: 10 ZRG1 CB-P (02) M Member Conflict: Cell Biology, Development and Aging

Science Outreach February 08-13, 2015

Niedernhofer Attended/speaker/session chair Gordon Research Conference "Mammalian DNA Repair"

Science Outreach February 19-20, 2015

Niedernhofer Reviewer, NIA/NIH Study Section Cellular Mechanisms of Aging and Development

Science Outreach January 29-31, 2015

Niedernhofer Attended/presented at PPG "Cell autonomous and nonautonomous mechanisms of aging", University of Pittsburgh

Science Outreach June 02-03, 2015

Niedernhofer Reviewer NIA/NIH K99 Biology of Aging Review Committee B study section

Science Outreach May 01-04, 2015

Niedernhofer Host Dr. Simon Watkins Chair of Cell Biology, University of Pittsburgh

Science Outreach May 31-June 01, 2015

Niedernhofer Attended "FASEB Animals in Research and Education Subcommittee Meeting; Science Policy Symposium; and Board of Directors Meeting", Arlington, VA

Science Outreach Monthly Niedernhofer Conference call with Geropathology Network Science Outreach Sept. 11-14,

2014 Niedernhofer Hosted Dr. Wim Vermeulen from Erasmus

Medical Center, Rotterdam, NL

Science Outreach Sept.15-16, 2014

Niedernhofer Attended FASEB Board of Directors Meeting, Arlington, VA

Science Outreach yearlong Patrick Griffin Elected Treasurer, ASMS (American Society of Mass Spectrometry)

Science Outreach yearlong Patrick Griffin Molecular Pharmacology Editorial Board member

Science Outreach yearlong Patrick Griffin JMB Editorial Board Member Science Outreach yearlong Patrick Griffin DMP Study Section Member Science Outreach yearlong Patrick Griffin CDRD (Centre for Drug Research and

Development) Board Member

Science Outreach Yearlong Donald Phinney Editor - Cytotherapy Science Outreach Yearlong Donald Phinney Member - ISCT MSCT Committee