Vascular Biology and Therapeutics Program VBT Annual Report... · Vascular Biology and Therapeutics...

YALE UNIVERSITY SCHOOL OF MEDICINE

Vascular Biology and Therapeutics Program

Annual Report 2013-2014

Sc siRNA Ucp2 siRNA

On the Cover: Autophagy levels in mouse lung endothelial cells, as assessed by immunofluorescent ptfLC3 plasmid after Scrambled, control siRNA (Sc siRNA) or Ucp2 siRNA. Original magnification of all photomicrographs: ×40. Submitted by Patty Lee, MD

TABLE OF CONTENTS

Message from the Director .....................................................................................................1 Program Operations ...............................................................................................................1

VBT Steering Committee .................................................................................................1 Administrative Operations ..............................................................................................1 Program Faculty Membership ........................................................................................2

Program Activities ..................................................................................................................4 Seminar Series .................................................................................................................5 Retreat ...............................................................................................................................5

Yale-Cambridge Program in Cardiovascular Disease .........................................................5 Tissue Engineering Group .....................................................................................................5 VBT Research in Progress (RIP) Talks .................................................................................5 Research Accomplishments ..................................................................................................6 Appendices

The Thirteenth Annual VBT Retreat ............................................................................. 1-1 The Twelfth Annual Meeting of the Joint Yale-Cambridge Program in .................... 2-1

Cardiovascular Research

Vascular biology & therapeutics

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Vascular Biology and Therapeutics Program (VBT)

Annual Report 2013 – 2014 Message from the Director The past academic year has been filled with exciting scientific developments with many VBT faculty publishing in top notch journals in 2013. Clearly this is a testimony to the excellence and high quality of work here at Yale within the VBT program. VBT membership is growing and now includes new members: Carlos Fernandez-Hernando, Ph.D. and Yajaira Suarez, Ph.D.

The interactions of VBT with the Cardiovascular Research Center (CVRC) growing with continued collaborations. Finally, I would like to applaud Themis Kyriakides who have done a wonderful job coordinating our joint seminar series and we appreciate their efforts. PROGRAM OPERATIONS VBT Steering Committee The Steering Committee serves as the principal advisory and leadership group for the program for the program. The current membership of the Steering Committee is listed in Table 1. Administrative Operations Ms. Carol Muzzey serves as the Program Manager and is assisted by Ms. Diane Strumpf. The program is served by the Central Administration Business Office.

VBT Steering Committee Jeffrey R. Bender, M.D., Robert I Levy Professor of Medicine (Cardiology) and Professor of Immunobiology; Associate Chief of Cardiovascular Medicine Alfred L.M. Bothwell, Ph.D., Professor of Immunobiology and Director of Graduate Studies Anne Eichmann, Ph.D., Professor Medicine (Cardiology) and Professor of Cellular and Molecular Physiology Frank Giordano, M.D., Associate Professor of Internal Medicine (Cardiology) Themis Kyriakides, Ph.D. Associate Professor of Pathology and of Biomedical Engineering Joseph A. Madri, M.D., Ph.D., Professor of Pathology and Director of Education Laura Niklason, M.D., Ph.D., Professor Anesthesia and Biomedical Engineering Jordan S. Pober, M.D., Ph.D., Bayer Professor of Translational Medicine; Director, Human and Translational Immunology Program; Vice-Chair Department of Immunobiology for the Section of Human and Translational Immunology Nancy H. Ruddle, Ph.D., Professor Emeritus of and Senior Research Scientist in Epidemiology W. Mark Saltzman, Ph.D., Goizueta Foundation Professor of Biomedical Engineering, Chemical & Environmental Engineering and Physiology William C. Sessa, Ph.D., Director Vascular Biology & Therapeutics, Professor and Vice Chair of Pharmacology and Professor of Medicine (Cardiology) Martin Schwartz, Ph.D., Robert W. Berliner Professor of Medicine (Cardiology) and Professor of Biomedical Engineering and of Cell Biology Michael Simons, M.D., RW Berliner Professor of Medicine and Cell Biology, Director of Yale Cardiovascular Research Center George Tellides, M.D., Ph.D., Professor of Surgery (Section of Cardiac Surgery) and of Investigative Medicine; Chief of Cardiothoracic Surgery, Veterans Affairs Medical Center

VASCULAR BIOLOGY AND THERAPEUTICS ANNUAL REPORT 2013 – 2014

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Program Faculty Membership All faculties at Yale with a significant interest in vascular biology and/or therapeutics are eligible to join VBT. VBT members in academic year 2013-2014 are:

Table 2 VBT Membership Jeffrey R. Bender, M.D., Robert I. Levy Professor of Medicine (Cardiology) and Professor of Immunobiology; Associate Chief, Cardiovascular Medicine Anton Bennett, Professor of Pharmacology and of Comparative Medicine; Co-Director, Program in Integrative Cell Signaling and Neurobiology of Metabolism; Director, BBS Minority Affairs Alfred L.M. Bothwell, Ph.D., Professor of Immunobiology and Director of Graduate Studies Demetrios Braddock, MD, PhD, Associate Professor of Pathology; Medical Director, Precipio Diagnostics David A. Calderwood, Ph.D., Associate Professor of Pharmacology and of Cell Biology Hyung J Chun, M.D., Assistant Professor of Medicine (Cardiology); Director, YPB Fellows' Cardiovascular Clinic Alan Dardik, M.D., Ph.D, Professor of Surgery (Vascular); Chief, Vascular Surgery, VA Connecticut Healthcare Systems, West Haven, CT Barbara E. Ehrlich, Ph.D., Professor of Pharmacology and of Cellular and Molecular Physiology Anne Eichmann, Ph.D., Professor of Medicine (Cardiology) and Professor of Cellular and Molecular Physiology Tarek Fahmy, Ph.D., Associate Professor of Biomedical Engineering and of Immunobiology Carlos Fernandez-Hernando, Ph.D., Associate Professor of Comparative Medicine Richard Flavell, Ph.D., FRS, Sterling Professor of Immunobiology; Investigator, Howard Hughes Medical Institute; Department Chair, Immunobiology Arnar Geirsson, M.D., Clinical Instructor in Surgery (Section of Cardiac Surgery); Director, Minimally Invasive Cardiac Surgery Frank J. Giordano, M.D., Associate Professor of Medicine (Cardiology) Daniel R. Goldstein, M.D., Professor of Medicine (Cardiology) and of Immunobiology Daniel Greif, M.D., Assistant Professor Medicine (Cardiology) Jaime Grutzendler, M.D., Associate Professor of Neurology and of Neurobiology; Director, Yale Center for Experimental Neuroimaging (YCEN) Murat Gunel, M.D., FACS, FAHA, Nixdorff-German Professor of Neurosurgery and Professor of Genetics and of Neurobiology; Co-Director, Yale Program on Neurogenetics; Director, Neurovascular Surgery Karen Hirschi, Ph.D., Professor of Medicine (Cardiology) Jay Humphrey, Ph.D., John C. Malone Professor of Biomedical Engineering John Hwa, M.D., Ph.D., Associate Professor of Medicine (Cardiology); Director of Cardiovascular Pharmacogenetics Yasuko Iwakiri, Ph.D., Assistant Professor of Medicine (Digestive Disease) Suk-Won Jin, Ph.D., Associate Professor (Adjunct) of Medicine (Cardiology) Martin S. Kluger, Ph.D., Research Scientist in Immunobiology Diane Krause, M.D., Ph.D., Professor of Laboratory Medicine, of Cell Biology and of Pathology; Assoc. Director, Yale Stem Cell Center; Assoc. Director, Transfusion Medicine Service; Medical Director, Clinical Cell Processing Laboratory; Medical Director, Advanced Cell Therapy Laboratory Sanjay Kulkarni, M.D., FACS, Associate Professor of Surgery (Transplant) and of Medicine (Nephrology) Themis Kyriakides, Ph.D., Associate Professor of Pathology and of Biomedical Engineering Patty J. Lee, M.D., Associate Professor of Medicine (Pulmonary) Joseph A. Madri, M.D., Ph.D., Professor of Pathology and Director of Education Arya Mani, M.D., Associate Professor of Medicine (Cardiology) and of Genetics; Director, Cardiovascular Genetics Program


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Kathleen A. Martin, Ph.D., Associate Professor of Medicine (Cardiology); Associate Professor of Pharmacology Laura R. Ment, M.D., Professor of Pediatrics (Neurology) and of Neurology; Associate Dean for Admissions and Financial Aid Wang Min, Ph.D., Professor of Pathology Stefania Nicoli, Ph.D., Assistant Professor of Medicine (Cardiology) Laura E. Niklason, M.D., Ph.D., Professor of Anesthesiology and Biomedical Engineering Jordan S. Pober, M.D., Ph.D., Bayer Professor of Translational Medicine; Director, Human and Translational Immunology Program; Vice-Chair, Dept. of Immunobiology for the Section of Human and Translational Immunology Yibing Qyang, Ph.D., Assistant Professor of Medicine (Cardiology) and of Pathology; Section of Cardiovascular Medicine Nancy Hartman Ruddle, Ph.D., Professor Emeritus of and Senior Research Scientist in Epidemiology (Microbial Diseases) Mehran M. Sadeghi, M.D., Associate Professor of Medicine (Cardiology) W. Mark Saltzman, Ph.D., Goizueta Foundation Professor of Biomedical Engineering, Chemical & Environmental Engineering & Physiology Martin Schwartz, Ph.D., Robert W. Berliner Professor of Medicine (Cardiology) and Professor of Biomedical Engineering and of Cell Biology William C. Sessa, Ph.D., Vice Chairman, Pharmacology; Director, Vascular Biology & Therapeutics Program; Alfred Gilman Professor of Pharmacology and Professor of Medicine (Cardiology) Michael Simons, M.D., Robert W. Berliner Professor of Medicine (Cardiology) and Professor of Cell Biology; Director, Yale Cardiovascular Research Center Albert J. Sinusas, M.D., Professor of Medicine (Cardiology) and of Diagnostic Radiology; Director, Translational Research Imaging Center; Director, Cardiovascular Imaging Jeffrey L. Sklar, M.D., Ph.D., Professor of Pathology and of Laboratory Medicine; Director, Molecular Diagnostics Program; Director, Molecular Genetics Pathology Fellowship; Director, Molecular Tumor Profiling Laboratory; Director of Molecular and Genomic Pathology Edward A. Snyder, M.D., Professor of Laboratory Medicine; Associate Chair, Clinical Affairs (Therapeutic); Director, Apheresis/Transfusion Service; Director, Blood Bank; Director of Membership, Yale Cancer Center; Editor, Lab News Bing Su, Ph.D., MPH, Associate Professor of Immunobiology Yajaira Suarez, Ph.D., Assistant Professor of Comparative Medicine and of Pathology George Tellides, M.D., Ph.D., Professor of Surgery (Section of Cardiac Surgery) and of Investigative Medicine; Chief of Cardiothoracic Surgery, Veterans Affairs Medical Center Jean-Leon Thomas, Ph.D., MSc, Associate Professor of Neurology Daniela Tirziu, Ph.D., Research Scientist in Medicine (Cardiology) Agnès M C Vignery DDS, PhD, Senior Research Scientist in Orthopaedics and Rehabilitation and in Cell Biology Dianqing (Dan) Wu, Ph.D., Professor of Pharmacology Lawrence H. Young, M.D., Professor of Medicine (Cardiology) and of Cellular and Molecular Physiology; Vice-Chairman, Department of Medicine Jun Yu, M.D., Research Scientist in Medicine (Cardiology)


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Table 3 – VBT 2012-2013 Seminar Series

SEPTEMBER 2013 Naftali Kaminski, M.D., Yale University, Boehringer-Ingelheim Endowed Professor of Internal Medicine, Chief of Pulmonary, Critical Care and Sleep Medicine "Coding and non-coding RNAs in lung fibrosis

OCTOBER 2013 Michael A. Choma, M.D.,Ph.D., "Optical Imaging of Microscale Cardiovascular Physiology, Assistant Professor, Departments of Diagnostic Radiology, Pediatrics, and Biomedical Engineering, Yale University

NOVEMBER 2013 Harry Ischiropoulos, Ph.D., Gisela and Dennis Alter Research Professor of Pediatrics and Pharmacology, Perelman School of Medicine, University of Pennsylvania, "Landscapes of endogenous cysteine proteomes: functional regulation of metabolism by S-nitrosylation" Valerie Horsley, Ph.D., Yale University, Maxine F. Singer Assistant Professor of Molecular, Cellular & Developmental Biology "Understanding the dermal macroenvironment in the skin”

DECEMBER 2013 Dianna Milewicz, M.D. Ph.D., President George Bush Chair in Cardiovascular Medicine, Professor, Division of Medical Genetics, Vice Chair, Department of Internal Medicine, The University of Texas-Houston Medical School, “Molecular Pathways to the Diverse and Diffuse Vascular Diseases Associated with Mutations in Smooth Muscle Alpha-Actin (ACTA2)” Jane E. Freedman, M.D. , Professor of Medicine, Director, Translational Research UMass Memorial Heart & Vascular Center University of Massachusetts Medical School, "Circulating RNA and Platelets in Vascular Homeostasis and Disease” Barry London, M.D., Ph.D., Professor of Medicine Potter-Lambert Chair, Director, Division of Cardiovascular Medicine, Co-Director, Cardiovascular Research Center, University of Iowa, "Sodium Channel Modifiers: From Brugada Syndrome to Heart Failure”

JANUARY 2014 Rong Fan, Ph.D., Assistant Professor, Biomedical Engineering, Yale University, "de novo bioengineering of large-scale, perfusable, functional endothelialized microvessel networks on a chip” Demetrios Braddock, M.D., Ph.D , Associate Professor of Pathology, Yale University, "Ectonucleotide Pyrophosphatases/Phosphodiesterases: A new class of vascular bound enzymes implicated in central hemostasis and thrombotic stroke.”

FEBRUARY 2014 Christopher C.W. Hughes, Ph.D., Professor and Chair, Department of Molecular Biology and Biochemistry, University of California Irvine, "The angiogenic microenvironment: probing the cellular and molecular interactions that regulate vascular sprouting.” Andre Terzic,M.D., Ph.D , Michael S. and Mary Sue Shannon Family Director, Center of Regenerative Medicine, Marriott Family Professor of Cardiovascular Research, Professor of Medicine and Pharmacology, Medical Genetics, "Cardiac Allostasis in Health and Disease”, Mayo Clinic, Rochester, Minnesota Amin S. Ghabrial, Ph.D., Assistant Professor of Cell and Developmental Biology, University of Pennsylvania School of

PROGRAM ACTIVITIES Seminar Series The VBT Monday afternoon seminars continue to serve as an intellectual focus of the vascular biology community at Yale. The series also serves as venue for assistance in the recruitment of faculty with research in vascular biology to various departments at Yale. The seminars are run by Dr. Themis Kyriakides. A list of seminar speakers and their titles are shown in Table 3.


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Medicine, "Tip cells and seamless tubes: morphogenesis and connectivity.”

MARCH 2014 Filip Swirski, Ph.D., Assistant Professor of Radiology, Director, Cell Core, Harvard University, "Leukocyte communication and the growth of atherosclerotic lesions” Mary Dickinson, Ph.D, Professor, Department of Molecular Physiology and Biophysics, Baylor College of Medicine, "Quantitative imaging reveals mechanisms underlying vessel remodeling: implications for animal development and tissue engineering” Beth A. Kozel, M.D.,Ph.D, Assistant Professor of Pediatrics in Genetics and Genomic Medicine, Washington University School of Medicine St. Louis, "Of mice and men; Modifiers of vascular disease in Williams syndrome”

APRIL 2014 Joseph Miano, Ph.D., Associate Professor of Medicine, Associate Director of the Cardiovascular Research Institute of Rochester, University of Rochester New York "Mining the Human Genome/Transcriptome for Functional Noncoding Sequences” Jean Philip Gratton, Ph.D., Professor and Chair Department of Pharmacology, Faculty of Medicine, University of Montreal, “Signaling at endothelial cell junctions: Implications in angiogenesis”

MAY 2014 Peter Hordijk, Ph.D., Head, Dept. Molecular Cell Biology Sanquin Laboratory, Academic Medical Center Swammerdam Institute for Life Sciences, University of Amsterdam, “Endothelial signaling in Leukocyte transmigration” Ken Poss, Ph.D., Professor, Cell Biology, Duke University, “Source and stimuli for heart regeneration”

Retreat The annual retreat continues to be an extremely popular activity, bringing together over one hundred ninety scientists from the laboratories of VBT faculty members. This past year, the retreat was held on October 18, 2013 at the Grace Murray Hopper Auditorium, West Campus. The retreat continued the poster session competition with prizes for the best posters by a graduate student and a post-doctoral fellow.

The Keynote Address at this years’ retreat was Ralf H. Adams, Ph.D., Professor, University of Muenster, Director Max Planck Institute for Molecular Biomedicine, “New Branches on the vascular tree: vessel heterogeneity and functional specialization”. The retreat Program is listed in Appendix 1.

Yale-Cambridge Program in Cardiovascular Disease

The research alliance with Cambridge has continued as an important activity, with 30 faculty from Cambridge visiting Yale in September 2013 for a two day scientific meeting. The program for this retreat is listed in Appendix 2. Tissue Engineering Group This biweekly forum, sponsored by VBT and organized by Dr. Themis Kyriakides, brings together investigators from Yale Medical School and Yale’s central campus to exchange updates in research in progress and to foster new research collaborations. VBT Research in Progress (RIP) Talks A monthly series organized by Marty Kluger, Ph.D. and Jun Yu, M.D. featuring presentations by graduate students and postdoctoral fellows working in VBT laboratories.


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Jeffrey R. Bender, M.D. Robert I. Levy Professor of Medicine (Cardiology) and Professor of Immunobiology; Associate Chief Cardiovascular Medicine 1. Overall Goal(s) of the Research Program of the Laboratory: My laboratory has had a longstanding interest in inflammation and immunity, as they relate to vascular physiology and pathology. The interactions between mononuclear leukocytes and endothelial cells play major roles in atherogenesis, acute and chronic manifestations of atherosclerosis, angiogenesis and allograft rejection. We have extended these studies to evaluating effects of ovarian steroid hormones on endothelial function. The work is performed at the cellular, molecular, and pre-clinical animal model levels. 2. Publications: (Publications July 1, 2013– June 30, 2014) Morrison AR, Yarovinsky TO, Young BD, Moraes F, Ross TD, Ceneri N, Zhang J, Zhuang ZW, Sinusas

AJ, Pardi R, Schwartz MA, Simons M and Bender JR: Chemokine-coupled β2 integrin-induced macrophage Rac2-Myosin IIA interaction regulates VEGF-A mRNA stability and arteriogenesis. J Exp Med. 2014 Sep 22;211 (10) :1957-68. Epub 2014 Sep 1. PMID: 25180062

Kim KH, Young BD and Bender JR: Endothelial estrogen receptor isoforms and cardiovascular disease. Mol Cell Endocrinol. 2014 May 25;389 (1-2) :65-70. Epub 2014 Feb 11. PMID: 24530925

Mehra VC, Jackson E, Zhang XM, Jiang XC, Dobrucki LW, Yu J, Bernatchez P, Sinusas AJ, Shulman GI, Sessa WC, Yarovinsky TO and Bender JR: Ceramide-activated phosphatase mediates fatty acid-induced endothelial VEGF resistance and impaired angiogenesis. Am J Pathol. 2014 May;184 (5) :1562-76. Epub 2014 Mar 5. PMID: 24606881

Chau KH, Bender JR and Elefteriades JA: Silver lining in the dark cloud of aneurysm disease. Cardiology. 2014;128 (4) :327-32. Epub 2014 Jun 17. PMID: 24942201


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Anton M. Bennett, Ph.D. Professor of Pharmacology

1. Overall Goal(s) of the Research Program of the Laboratory: The broad research interests of this laboratory are to define the molecular mechanisms, physiological and pathophysiological roles of the protein tyrosine phosphatase (PTP) family of enzymes. Using mouse genetic approaches we are studying how PTPs are involved in regulating metabolism. This work suggests new modes of control through which PTPs participate in metabolic signaling and potentially the progression of obesity and diabetes. We are also investigating the pathophysiological mechanisms of the PTPN11 gene, which encodes for the PTP, SHP-2. PTPN11/SHP-2 mutations are found in ~50% of Noonan syndrome cases, which represents the largest non-chromosomal cause of congenital heart disease. SHPThe broad goal of this project is to identify the signaling mechanisms induced by the mutant form of PTPN11SHP-2 that leads to congenital heart disease. 2. Specific Research Accomplishments in the last 12 months: We have identified novel targets involved in the pathogenesis of congenital heart disease. These targets are regulated by SHP-2 which is aberrantly regulated in Noonan syndrome. 3. Significance of Key Findings Relevant for the Mission of VBT: The identification of protein zero-related (PZR) as a potential signaling target of PTPN11/SHP-2 in the development of Noonan syndrome may offer new insight in to therapies for the treatment of heart disease. 4. Publications: (Publications July 1, 2013 – June 30, 2014) Lee, H. and Bennett, AM. (2013) Receptor Tyrosine Kinase-Receptor Tyrosine Phosphatase Substrate

Screen Identifies EphA2 as a LAR Substrate In Cell Migration, Mol. Cell. Biol., 33: 1430-1441. Shi, H., Verma, M., Zhang, L., Dong, C., Flavell, R. A. and Bennett, AM (2013), Improved regenerative

myogenesis and muscular dystrophy in mice lacking MKP-5, J. Clin. Invest., 123: 2064-2077. Amanda N. Kallen, Jie Xu, Chong Qiao, Clémence Martinet, Jing Ma, Xiao-Bo Zhou, Lei Yan, Lingeng Lu,

Chaochun Liu, Jae-Sung Yi, Haifeng Zhang, Min Wang, Richard I. Gregory, Anton M. Bennett, Ye Ding, Anne Gabory, Luisa Dandolo & Yingqun Huang (2013), Imprinted H19 lncRNA antagonizes let-7 miRNAs in muscle, Mol. Cell, 52: 101-12.

Overman, J. P., Lee, J-S., Bonetti, M., Soulsby, M., Preisinger, C., Stokes, M. P., Hui, L., Silva, J. C., Heck, A. J. R., Kontaridis, M. I., den Hertog, J., Bennett, AM (2014) PZR coordinates Shp2 phosphatase-independent Noonan and LEOPARD syndrome signaling in zebrafish and mice, Mol. Cell. Biol., 34: 2874-89.

Lawan, A. and Bennett, AM (2014) MAP Kinase phosphatases emerge as new players in metabolic regulation, In Protein Tyrosine Phosphatase Control of Metabolism,

K. Bence ed. (Springer), Ch. 12. Lee, H. and Bennett, AM (2014) Identification of Receptor protein tyrosine phosphatases (RPTPs) as

regulators of Receptor Tyrosine Kinases (RTK) using an RPTP siRNA-RTK substrate screen, Methods in Molecular Biology, S. Germano ed., in press.

Lee, H., Lawan, A., Yi, J-S, Min, K. and Bennett, AM (2014) Mining the functions of protein tyrosine phosphatases in Health and Disease, Seminars in Cell and Developmental Biology, in press.


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Alfred L.M. Bothwell, Ph.D. Professor, Dept. of Immunobiology

1. Overall Goal(s) of the Research Program of the Laboratory: The overall goals are to understand the mechanisms utilized by regulatory lymphocyte populations to affect the development of vascular disease, autoimmunity and cancer. 2. Specific Research Accomplishments in the last 12 months: The laboratory has now published an in vivo model of the earliest phase of type 1 diabetes utilizing islet autoantigen specific T cells that demonstrates specific homing and infiltration of islets. Current efforts are to extend this model to include autoantigen specific human CD8 T cells. Several humanized mouse models of cancer are under development. The hope is that such models can be of value in testing personalized therapeutic strategies with a patient tumor and lymphocytes in the same biologic model. 3. Significance of Key Findings Relevant for the Mission of VBT: Ever since we first cloned human CD59, the terminal membrane attack complex inhibitor, 25 years ago we have been seeking meaningful molecular connections between the coagulation system and the immune system. In the last few months we have finally succeeded in this goal by showing that the major source of wnt antagonist Dkk1 in peripheral blood is platelets. There are remarkable immunoregulatory consequences of Dkk1 on the differentiation and proliferation of T cell subsets. This suggests a very significant role for platelets in regulating immunity in general and raises the prospect of new therapeutic targets. 4. Publications: (Publications July 1, 2013– June 30, 2014) Kobsa, S., Kristofik, N.J., Sawyer, A.J., Bothwell, A.L.M., Kyriakides, T.R. and W. Mark Saltzman, W.M.

(2013). An electrospun scaffold integrating nucleic acid delivery for treatment of full thickness wounds. Biomaterials, (epub ahead of print) doi:10.1016/j.biomaterials.2013.02.016 PMCID: PMC3625647.

Senejani, A., Liu, Y., Kidane, D., Maher, S.E., Zeiss, C.J., Park, H.-J., Kashgarian, M., McNiff, J.M., Bothwell, A.L.M. and Sweasy, J.B. (2014). Mutation of DNA Polymerase Beta Results in Lupus, Cell Reports 6:1-8. PMCID: PMC3916967.

Kidane, D., Chae, W.J., Czochor, J., Eckert, K.A., Glazer, P.M., Bothwell, A.L.M., Sweasy, J.B. (2014). Links between DNA repair and inflammation. Crit. Rev. Biochem. Mol. Biol., 49:116-139.

Viehmann Milam, A.A., Maher, S.E., Gibson, J.A., Lebastchi, J., Wen, L., Ruddle, N.H., Herold, K.C. and Bothwell, A.L.M. (2014). A Humanized Mouse Model of Autoimmune Insulitis. Diabetes, 63:1712-1724.

Park, H.-J., Choi, J.-Y., Senejani, A.G., Tobiasova, Z., Kim, D.-H., * Bothwell, A.L.M. and * Choi, J.-M., (2014).The nuclear receptor PPARγ regulates T cell tolerance and germinal center formation via follicular helper T cells. PLoS ONE 9(6): e99127. *joint senior authors.

Ehrlich, A., Castilho, T., Goldsmith-Pestana, K., Chae, W.-J., Bothwell, A.L.M., Sparwasser, T., and Diane McMahon-Pratt. D. (2014). The immunotherapeutic role of regulatory T cells in Leishmania (Viannia) panamensis infection. J. Immunol., in press. PMID: 25098291.

Park, J.-J., Ko, J.S., Shin, Y, Cho, J.Y., Oh, H.-a., Bothwell, A.L.M., Lee, S.-K. (2014). Intranuclear interactomic inhibition of FoxP3 suppresses functions of Treg cells. Biochem. Biophys. Res. Commun., 451:1-7.

Park, T.-Y., Sung-Dong Park, S.-D., Cho, J.-Y., Moon, J.-S., Kim, N.-Y., Lee, S.-W., Morio, T., Bothwell, A.L.M. and Lee, S.-K. RORγt-specific transcriptional interactomic inhibition suppresses autoimmunity associated with TH17 cells. Submitted.

Chae, W.-J., Ehrlich, A., Chan, P., Henegariu, O., Hao, L., Goldsmith-Pestana, K., Tang, W.-H., Texeira, A.M., Kim, S.-T., Park, J.-H., Maher, S.E., Hwa J., Rothlin, C.V., McMahon-Pratt, D. and Bothwell, A.L.M. Wnt antagonist Dickkopf-1 from platelets promotes type 2 inflammatory diseases. Submitted.


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David A Calderwood, Ph.D. Associate Professor of Pharmacology and of Cell Biology

1. Overall Goal(s) of the Research Program of the Laboratory: The broad goal of research in the Calderwood lab is to understand the molecular basis of signaling to and from integrin adhesion receptors. We use a combination of structural, biochemical and cell biological approaches to investigate how protein-protein interactions regulate integrin activation state, provide links from integrins to the actin cytoskeleton, control cell migration and morphogenesis, and assemble into signaling complexes. More recently, in collaboration with the Boggon lab at Yale, we have also become interested in cerebral cavernous malformation (CCM) proteins and their interactions. 2. Specific Research Accomplishments in the last 12 months: Over the past year we have continued our longstanding interest in integrin activation with a particular focus on the roles of kindlins in integrin activation and signalling. Specifically we have revealed the importance of kindlin interactions with integrin-linked kinase both for targeting of kindlins to integrin-rich focal adhesions and for kindlin-mediated integrin activation. We have also reported initial characterization of kindlin-migfilin interactions and contributed to collaborative studies on the roles of talin in the kidney and assessed consequences of loss of dynamin loss on integrin activation in endothelial cells. In addition, together with crystallographers in the Boggon lab, we have extended our investigations of the functional consequences of disruption of specific protein-protein interactions within the cerebral cavernous malformation (CCM) protein network. 3. Significance of Key Findings Relevant for the Mission of VBT: Integrin activation and signaling is critical for platelet aggregation, angiogenesis and leukocyte trafficking and the links between integrins and actin play key mechano-signaling roles during cell- adhesion and tissue formation with clear relevance to the vascular system. Our recent studies into CCM proteins also establish the importance of CCM complex formation in endothelial cell function. 4. Publications: (Publications July 1, 2013– June 30, 2014) Ra zinia Z.*, Baldassarre M.*, Cantelli G., and Calderwood D.A. (2013) ASB2α an E3 ubiquitin ligase

specificity subunit, regulates cell spreading and triggers proteasomal degradation of filamins by targeting the filamin calponin homology 1 domain. J. Biol. Chem. 288:32093-105

Brahme N.N., Harburger D.S., Kemp-O’Brien K., Stewart R., Raghavan S., Parsons M., and Calderwood D.A. (2013) Kindlin binds migfilin tandem LIM domains and regulates migfilin focal adhesion localization and recruitment dynamics J. Biol. Chem. 288:35604-16.

Draheim K.M., Fisher O.S., Boggon T.J. and Calderwood D.A. (2014) Cerebral Cavernous Malformation (CCM) Proteins at a Glance. J. Cell Sci. 127:701-707

Morse E.M, Brahme N.N. and Calderwood D.A. (2014) Integrin cytoplasmic tail interactions. Biochemistry 53:810-820.

Lee M., Skoura A., Park E.J., Landskroner-Eiger, S., Jozsef L., Luciano A.K., Murata T., Pasula S., Dong

Y., Bouaouina M., Calderwood D.A., Ferguson S.M, De Camilli P., and Sessa W.C. (2014) Dynamin-2 regulation of integrin endocytosis, but not VEGF signaling, is critical for developmental angiogenesis. Development 141:1465-1472.

Tian X.*, Kim J.J.*, Monkley S.M., Gotoh N., Nandez R., Soda K., Inoue K., Balkin D.M., Hassan H., Son S.H., Lee Y., Moeckel G., Calderwood D.A., Holzman L.B., Critchley D.R., Zent R., Reiser J., and Ishibe S. (2014) Podocyte-associated talin1 is critical for glomerular filtration barrier maintenance. J. Clin. Invest. 124:1098-1113.


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Hyung J. Chun, M.D. Assistant Professor of Medicine (Cardiology); Director YPB Fellows’ Cardiovascular Clinic 1. Overall Goal(s) of the Research Program of the Laboratory: The Chun Laboratory studies a number of signaling processes that regulate vascular function in health and disease. Research is focused on identifying signaling pathways that play key roles in the context of cardiovascular disease and development, including pulmonary arterial hypertension, atherosclerosis and cardiovascular development. 2. Specific Research Accomplishments in the last 12 months: We continue to investigate the signaling mechanisms that are involved in pathogenesis of multiple cardiovascular disease processes, including pulmonary arterial hypertension, atherosclerosis, and diabetic endothelial dysfunction. Moreover, we are actively investigating the crosstalk of various G protein coupled receptor signaling pathways that are intricately involved in vascular development. Key highlights include:

a) Demonstration of novel transcriptional regulation of endothelial microRNAs that are downregulated in pulmonary arterial hypertension, which can be restored via administration of a selective histone deacetylase inhibitor

b) Demonstration of crosstalk between APLNR and CXCR4 GPCRs, two GPCRs that are selectively expressed in the vascular endothelium, via a novel microRNA mediated mechanism

c) Demonstration that endothelial apelin signaling is key to regulation of fatty acid uptake and can be targeted for treatment of diabetes mellitus.

3. Significance of Key Findings Relevant for the Mission of VBT: We continue to explore vascular signaling pathways that are important in both vascular disease states as well as maintenance of vascular homeostasis. Better understanding of these signaling mechanisms will provide greater insights into the mechanisms of disease pathogenesis, as well as identify novel venues for potential therapies aimed at treating vascular diseases such as atherosclerosis and pulmonary arterial hypertension. 4. Publications: (Publications July 1, 2013-June 30, 2014) Kim, JD, Kang, Y, Kim, J, Papangeli, I, Kang, H, Wu, J., Fischer, J, Park, H, Nadelmann, E, Rockson, SG,

Ober, EA, Chun, H.J.,* Jin, SW* (*co-corresponding authors) (2014). Essential Role of Apelin Signaling during Lymphatic Development in Zebrafish, ATVB, 34:338-345, featured in editorial in ATVB, Vol, 34; P 239-241

Mehrotra, D., Wu, J., Papangeli, I., Chun, H.J., Endothelium as a gatekeeper of fatty acid transport, Trends in Endocrinology and Metabolism, 25(2): 99-106, 2014


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Alan Dardik, M.D., Ph.D. Professor of Surgery (Vascular)

1. Overall Goal(s) of the Research Program of the Laboratory: The Dardik laboratory continues to study the healing and function of blood vessels as used in patients having vascular bypass surgery as well as arteriovenous fistulae (AVF). We are currently trying to understand the fundamental molecular mechanisms by which vein graft adaptation and AVF maturation result in positive remodeling and successful adaptation to the arterial environment, yet often proceed, in the long-term, to neointimal hyperplasia and graft and AVF failure. 2. Specific Research Accomplishments in the last 12 months: The laboratory continues to study the mechanisms by which Eph-B4 controls venous adaptation to the arterial circulation; we received a new VA MERIT award to fund research on AVF maturation. We have continue to characterize our model of the mouse AVF and have shown its similarity to human AVF maturation. We continue to collaborate closely with several other labs in the VBT programs, as well as with collaborators at UCL. The Dardik Lab website (http://medicine.yale.edu/lab/dardik/index.aspx) has finally gone live and highlights our place within VBT.

3. Significance of Key Findings Relevant for the Mission of VBT: Our laboratory continues to identify mechanisms of venous remodeling and adaptation to the arterial circulation.

4. Publications: (Publications July 1, 2013– June 30, 2014) Yamamoto K, Li X, Shu C, Miyata T, Dardik A. Technical aspects of the mouse aortocaval fistula. Journal

of Visualized Experiments Jul 11;77:e50449 (2013). Yamamoto K, Protack CD, Tsuneki M, Hall MR, Wong DJ, Lu DY, Assi R, Williams WT, Sadaghianloo N,

Bai H, Miyata T, Madri JA, Dardik A. The mouse aortocaval fistula recapitulates human arteriovenous fistula maturation. American Journal of Physiology – Heart and Circulatory Physiology 305(12):H1718-H1725 (2013).

Kondo Y, Jadlowiec CC, Muto A, Yi T, Protack C, Collins MJ, Tellides G, Sessa WC, Dardik A. The Nogo-B-PirB axis controls macrophage-mediated vascular remodeling. PLoS ONE 8(11):e81019 (2013).

Orozco–Sevilla V, Naftalovich R, Hoffmann T, London D, Czernizer E, Yang C, Dardik A, Dardik H. EGCG is a potent phytochemical inhibitor of intimal hyperplasia in the wire-injured carotid artery. Journal of Vascular Surgery 58(5):1360-1365 (2013).

Jadlowiec C, Dardik A. Shear stress and endothelial cell retention in critical lower limb ischemia. In: Gabriel EA, Gabriel SA. Inflammatory Response in Cardiovascular Surgery. Springer, London. p107-116 (2013).

Lu DY, Chen EY, Wong DJ, Yamamoto K, Protack CD, Williams WT, Assi R, Hall MR, Sadaghianloo N, Dardik A. Vein graft adaptation and fistula maturation in the arterial environment. Journal of Surgical Research 188:162-173 (2014).

Model LS, Hall MR, Wong DJ, Muto A, Kondo Y, Ziegler KR, Feigel A, Quint C, Niklason L, Dardik A. Arterial shear stress reduces Eph-B4 expression in adult human veins. Yale Journal of Biology and Medicine in press (2014).

Hall MR, Yamamoto K, Protack CD, Tsuneki M, Kuwahara G, Assi R, Brownson KE, Bai H, Madri JA, Dardik A. Temporal Regulation of Venous Extracellular Matrix Components during Arteriovenous Fistula Maturation. Journal of Vascular Access in press (2014).


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Anne Eichmann, Ph.D. Ensign Professor, Cardiovascular Research Department

1. Overall Goal(s) of the Research Program of the Laboratory: My laboratory studies vascular development, with particular interests in mechanisms that direct patterning and guidance. Specialized endothelial cells (EC) called tip cells located at the extremities of growing capillary sprouts mediate guided vascular patterning. Tip cells exhibit characteristic features, including extension of filopodia that explore the tip cell environment, lack of a lumen and a slow proliferation rate. Following behind tip cells, other EC termed stalk cells form the capillary lumen and proliferate. Our research is focused on molecular events guiding angiogenesis and arteriogenesis:

2. Specific Research Accomplishments in the last 12 months: We have identified various means of enhancing arteriogenesis in mice, including i) disruption of Dll4 Notch signaling, which leads to increased arterial branching and collateral artery formation, ii) expression of a constitutively active Raf1 construct that enhances ERK signaling, which results in formation of exuberant arterial vasculature, and iii) inactivation of PTP1B, a phosphatase targeting VEGFR2. These studies are consistent with a model whereby VEGF-driven ERK signaling controls arteriogenesis, and downstream Notch signaling serves to limit the extent of this process. Furthermore we have identified signals controlling innervation of arterial smooth muscle cells by sympathetic neurons. We have shown that arterial innervation by sympathetic nerve fibers is initiated during early postnatal development (postnatal day P2) in mice, after other visceral smooth muscle innervation is completed. The late temporal onset of arterial innervation compared to that of other smooth muscle targets suggests that postnatal arteries produce factors that initiate their innervation. We have identified signals controlling this process, including the axonal guidance molecule Netrin-1. Netrin-1 is expressed by arterial smooth muscle cells at the onset of innervation at P2. Function-blocking experimental approaches including cell type–specific deletion of the genes that encode Netrin-1 (Ntn-1 ) in smooth muscle cells and its receptor DCC (Deleted in Colorectal Cancer) in sympathetic neurons leads to a selective reduction in arterial innervation without affecting o t h e r s y m p a t h e t i c t a r g e t s . Reduced arterial innervation, in turn, leads to defective vasoconstriction in adult Ntn -1 deficient animals. Mice carrying temporally inducible, smooth muscle cell–specific deletions of Netrin-1–encod ing Ntn-1 gene and sympathetic neuron–specific deletions of the DCC-encoding Dcc gene provide experimental models in which to study the contribution of sympathetic arterial innervation to blood pressure regulation and to investigate the role of sympathetic nerves in arterial development and vascular disease. 3. Significance of Key Findings Relevant for the Mission of VBT: Our findings provide new insight into fundamental mechanisms directing angiogenesis and arteriogenesis. 4. Publications: (Publications July 1, 2013– June 30, 2014) Cristofaro B, Shi Y, Faria M, Suchting S, Leroyer AS, Trinidade A, Duarte A, Zovein AC, Iruela-Arispe ML,

Nih LR, Kubis N, Henrion D, Loufrani L, Todiras M, Schleifenbaum J, Gollasch M, Zhuang ZW, Simons M, Eichmann A, le Noble F. Dll4-Notch signaling modulates arteriogenesis and functional recovery in arterial occlusion models in mice. Development 2013; 140: 1720-1729.

Deng Y, Larrivée B, Zhuang ZW, Atri D, Moraes F, Prahst C, Eichmann A, Simons M. Endothelial RAF1/ERK activation regulates arterial morphogenesis. Blood 2013; 121: 3988-3996, S1-9.

Simons M, Eichmann A. Lymphatics are in my veins. Science 2013; 341: 622-624. Eichmann A, Simons M. Need glucose to sprout: local metabolic control of angiogenesis. Embo Mol. Med.

2013; 5: 1459-61. Atri D, Larrivee B, Eichmann A, Simons M. Endothelial Signaling and the Molecular Basis of

Arteriovenous Malformations. Cell Mol Life Sci. 2013; Sept. 28.


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Nie L, Guo X, Esmailzadeh L, Zhang J, Asadi A, Collinge M, Li X, Kim JD, Woolls M, Jin SW, Dubrac A,

Eichmann A, Simons M, Bender JR, Sadeghi MM. ESDN modulates angiogenesis and regulates VEGF signaling in endothelial cells. J. Clin. Invest. 2013; 123: 5082-5097.

Prahst C, Kasaai B, Moraes F, Jahnsen ED, Larrivee B, Villegas D, Pardanaud L, Pibouin-Fragner L,

Zhang F, Zaun HC, Eichmann A, Jones EAV. The homeobox transcription factor Hlx modulates yolk sac vascular remodeling in mouse embryos. ATVB 2014; 34: 1468-1476.

El Hallani S, Colin C, El Houfi Y, Idbaih A, Boisselier B, Marie Y, Ravassard P, Labussière M, Mokhtari K, Thomas JL, Delattre JY, Eichmann A, Sanson M. Tumor and endothelial cell hybrids participate in glioblastoma vasculature. Biomed Res Int. 2014; 827327.

Greif D, Eichmann A. Vascular Biology: Brain vessels squeezed to death. Nature 2014; 508: 50-1. Brunet I, Gordon E, Han J, Cristofaro B, Broqueres-You D, Liu C, Bouvrée K, Zhang J, del Toro R,

Mathivet T, Larrivée B, Jagu J, Pibouin-Fragner L, Pardanaud L, Machado MJC, Kennedy TE, Zhuang ZW, Simons M, Levy BI, Tessier-Lavigne M, Grenz A, Eltzschig H, Eichmann A. Netrin-1 controls sympathetic arterial innervation. J. Clin. Invest. 2014; 24: 3230-3240.

Lanahan AA, Lech D, Dubrac A, Zhang A, Zhuang ZW, Eichmann A, Simons M. PTP1b is a physiologic regulator of VEGF signaling in endothelial cells. Circulation 2014; June 30.


14

Carlos Fernández-Hernando, PhD Associate Professor of Comparative Medicine and Pathology

1. Overall Goal(s) of the Research Program of the Laboratory: The Fernández-Hernando laboratory studies the molecular regulation of lipoprotein metabolism and how its deregulation contributes to cardiometabolic diseases such as atherosclerosis. Our specific goals are to understand how microRNAs and other non-coding RNAs regulate cellular cholesterol metabolism and the identification of novel genes that controls cellular cholesterol metabolism using high-throughput screening approaches. . 2. Specific Research Accomplishments in the last 12 months: Discoveries over the past 12 months include: (1) Identification of miR-30c and miR-145 as posttranscriptional regulators of ABCA1 expression, a key transporter that regulates cellular cholesterol efflux and HDL biogenesis; (2) Identification of the RNA-binding protein HuR as a regulator of ABCA1 expression in macrophages and human hepatic cells;(3) Identification the role of miR-33 in controlling glucose metabolism and that long-term therapeutic silencing of miR-33 increases circulating triglyceride levels and hepatic lipid accumulation in mice; and (4) Demonstration that therapeutic silencing of miR-33 attenuates the progression of atherosclerosis in mice. 3. Significance of Key Findings Relevant for the Mission of VBT: Our research can form the basis of new therapies for treating dyslipidemia and cardiovascular related disorders including atherosclerosis. 4. Publications: (Primary Research Publications July 1, 2013– June 30, 2014) miR-30c lowers lipid synthesis and lipoprotein secretion to reduce hyperlipidemia and atherosclerosis. Soh

J, Iqpal J, Queiroz J, Fernández-Hernando C and Hussain MM. Nat Med. 2013. Jul;19(7):892-900. Therapeutic silencing of microRNA-33 in mice inhibits the progression of atherosclerosis. Rotllán N,

Ramirez C, Esau C and Fernández-Hernando C. Arterioscler. Thromb. Vasc. Biol. 2013. Aug;33(8): 1973-77.

MiR-33 regulates glucose production. Ramírez CM, Goedeke L, Rotllan N, Yoon JH, Mattison JA, Suarez Y, de Cabo R, Gorospe M, and Fernández-Hernando C. Mol Cell Biol. 2013. Aug;33(15):28911-202.

Impaired liver regeneration in Ldlr knockout mice is associated with an altered hepatic profile of cytokines, growth factors and lipids. Pauta M, Rotllan N, Vales-Lara F, Allen RM, Ford DA, Marí M, Jiménez W, Baldán A, Morales-Ruiz M, Fernández-Hernando C. J Hepatol. 2013. Oct;59(4):731-737.

MiR-155-/- mice are susceptible to dietary non-alcoholic hepatosteatosis. Miller AM, Nijjar J, Araldi E, Gilchrist DS, Asquith DL, Lavery CA, Ruzicska E, Fernández-Hernando C, McInnes IB and Kurowska-Stolarska M. 2013. PLoS One. 2013. Aug 21;8(8):e72324.

Curcumin promotes exosomes/microvesicles secretion, which attenuates lysosomal cholesterol traffic impairment. Canfran-Duque A, Pastor O, Quintana-Portillo R, Lerma M, De la Pena G, Martin-Hidalgo A, Fernandez-Hernando C, Lasuncion MA, Busto R. Mol Nutr Food Res. 2014. Apr;5894:687-697.

Improved repair of dermal wounds in mice lacking microRNA-155. van Solingen C, Araldi E, Chamorro-Jorganes A, Fernández-Hernando C and Suárez Y. J Cell Mol Med. 2014. March 17.

HuR regulates cholesterol efflux through post-transcriptional regulation of ABCA1. Ramírez CM, Lin CS, Abdelmohsen K, Goedeke L, Yoon JH, Madrigal-Matute J, Martin-Ventura JL, Penalva LO, Gorospe M, Fernández-Hernando C. J Lipid Res. 2014. Apr 11;55(6):1066-1076.

miR-143/145 deficiency protects against the progression of atherosclerosis in Ldlr-/- mice. Sala F, Aranda

JF, Rotllan N, Ramírez CM, Elia L, Condorelli G, Catapano AL, Fernández-Hernando C*, Norata DG*. (corresponding authors). Thromb Haemost. 2014. Jul 10;112(4). [Epub ahead of print].

Long-term therapeutic silencing of miR-33 increases circulating triglyceride levels and hepatic lipid accumulation in mice. Goedeke L, Salerno A, Ramírez CM, Guo L, Allen RM, Yin X, Langley SR, Esau C, Wanschel AC, Fisher EA, Suárez Y, Baldán A, Mayr M and Fernández-Hernando C. EMBO Mol Med. 2014. Jul 18. [Epub ahead of print].


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Richard Flavell, Ph.D. FRS, Sterling Professor of Immunobiology, Howard Hughes Medical Institute; Department Chair Immunobiology 1. Overall Goal(s) of the Research Program of the Laboratory: We have used knockout mice to investigate the role of an innate inflammatory triggering mechanism in regulation of Goblet cell biology and intestinal homeostasis. The inflammasome is an intercellular protein apparatus that can response to stress signals and activate an inflammatory response by mediating the maturation and secretion of inflammatory cytokines. Goblet cells are specialized mucus producing cells differentiated from intestinal epithelial stem cells and required for establishment of a protective mucus barrier between intestinal epithelial cells and commensal or pathogenic bacteria residing in the gut lumen. Due to the robust contribution of both the inflammasome apparatus and Goblet cells to the maintenance of intestinal homeostasis, we have set to investigate whether mucus production in Goblet cells is regulated by the inflammasome. 2. Specific Research Accomplishments in the last 12 months: Mucus production by goblet cells of the large intestine serves as a crucial antimicrobial protective mechanism at the interface between the eukaryotic and prokaryotic cells of the mammalian intestinal ecosystem. However, the regulatory pathways involved in goblet cell-induced mucus secretion remain largely unknown. Here, we demonstrate that the NLRP6 inflammasome, a recently described regulator of colonic microbiota composition and biogeographical distribution, is a critical orchestrator of goblet cell mucin granule exocytosis. NLRP6 deficiency leads to defective autophagy in goblet cells and abrogated mucus secretion into the large intestinal lumen. Consequently, NLRP6 inflammasome-deficient mice are unable to clear enteric pathogens from the mucosal surface, rendering them highly susceptible to persistent infection. This study identifies an innate immune regulatory pathway governing goblet cell mucus secretion, linking nonhematopoietic inflammasome signaling to autophagy and highlighting the goblet cell as a critical innate immune player in the control of intestinal host-microbial mutualism. 3. Significance of Key Findings Relevant for the Mission of VBT: The above findings led to the discovery of Nlrp6 as a key and non-redundant regulator of autophagy- mediated mucus granule exocytosis in goblet cells. Autophagy, granule secretion and protein secretion are, however, also important in regulating endothelial cell functions. Moreover, vascular activation is key in mediating immune cell infiltration into compromised tissues at the initial stages of inflammation. Thus, inflammasome components may directly or indirectly regulate vascular physiology, highlighting the relevance of these findings to the mission of VBT. 4. Publications: (Publications July 1, 2013– June 30, 2014) Wlodarska, M., C. A. Thaiss, R. Nowarski, J. Henao-Mejia, J. P. Zhang, E. M. Brown, G. Frankel, M.

Levy, M. N. Katz, W. M. Philbrick, E. Elinav, B. B. Finlay, and R. A. Flavell. 2014. NLRP6 Inflammasome Orchestrates the Colonic Host-Microbial Interface by Regulating Goblet Cell Mucus Secretion. Cell 156:1045-1059.

Gagliani N, Palm NW, de Zoete MR, Flavell RA. Inflammasomes and intestinal homeostasis: regulating and connecting infection, inflammation and the microbiota. Int Immunol. Jun 19. pii: dxu066 (2014) [Epub ahead of print] PubMed PMID: 24948595.

Henao-Mejia J, Elinav E, Thaiss CA, Flavell RA. Inflammasomes and Metabolic Disease. AnnuRev Physiol. 10;76:57-78. Feb. (2014) PMID: 24274736.


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Daniel R. Goldstein, M.D. Professor of Medicine (Cardiology) and of Immunobiology 1. Overall Goal(s) of the Research Program of the Laboratory: Understand how aging modifies inflammation and how inflammation alters organ Transplantation 2. Specific Research Accomplishments in the last 12 months:

Determine a novel pathway by which ER stress induces apoptosis in macrophages Discern novel mechanisms by which aging enhances atherosclerosis Disease novel pathway of inflammation after cardiac transplantation

3. Publications: (Publications July 1, 2013– June 30, 2014) Song Y, Shen H, Du W and Goldstein DR. Inhibition of x-box binding protein 1 expression reduces

tunicamycin-induced apoptosis in aged murine macrophages. Aging Cell 2013 (12): 794-801 Elpek, KG Cremasco V, Shen H, Goldstein DR, Monach PA. Turley SJ. The tumor microenvironment

shapes lineage, transcriptional, and functional diversity of infiltrating myeloid cells Cancer Immunology Research 2014 (7):655-67

Wong C and Goldstein DR. Impact of Aging on Antigen Presentation Function of Dendritic Cells Current Opinion in Immunology 2013 25(4):535-41. Review

Li W, Goldstein DR and Kreisel D Intravital 2 photon Imaging, leukocyte trafficking and the beating heart Trends in Cardiovascular Medicine 2013 S1050-1738(13)00062-5. Review

Shen H, Kreisel D and Goldstein DR. Processes of Sterile Inflammation. The Journal of Immunology 2013 191(6):2857-63. Review

Age-dependent dysregulation of innate immunity. Shaw AC, Goldstein DR and Montgomery RR. Nature Reviews Immunology 2013 (12):875-87. Review

Mori D, Kreisel D, Fullerton JN, Gilroy DW and Goldstein DR Inflammatory triggers of acute organ allografts Immunological Reviews 2014 258:132-144, Review


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Daniel M. Greif, M.D. Assistant Professor, Cardiovascular Section, Department of Medicine

1. Overall Goal(s) of the Research Program of the Laboratory: My laboratory investigates blood vessel morphogenesis, the maintenance of the adult blood vessel and diseases of the vasculature. We initially determined the patterns of smooth muscle and adventitial cell differentiation, proliferation and migration in the developing pulmonary arterial wall in the mouse and the role of the platelet derived growth factor pathway in these processes. Currently, we are studying the morphogenesis of the walls of the aorta and the intracerebral vasculature, and comparing and contrasting their development with that of the pulmonary artery. In addition, little is known about the maintenance of blood vessels, and we aim to determine the patterns and underlying mechanisms of cell turnover, proliferation and migration in the adult vessel wall. Moreover, diseases of the vasculature are thought to largely involve a recapitulation of developmental programs, and we are investigating animal models of vascular diseases that involve defective vascular walls, such as atherosclerosis, arterial stenosis, pulmonary artery hypertension and intracerebral hemorrhage. Finally, we are studying clinical samples obtained from patients with vascular diseases and relating them to our findings in animal models. 2. Specific Research Accomplishments in the last 12 months: A key pathological feature of pulmonary hypertension is the distal extension of smooth muscle cells to normally non-muscularized pulmonary arterioles, but underlying mechanisms are not defined. We recently identified that distal arteriole SMCs in hypoxia-induced pulmonary hypertension derive from pre-existing proximal pulmonary artery smooth muscle and undergo stereotyped stages of dedifferentiation, distal migration down the uncoated endothelial cell tube, proliferation and finally redifferentiation. We have also made seminal discoveries regarding blood-brain barrier formation and aortic wall development and disease (e.g. supravalvular aortic stenosis and atherosclerosis). 3. Significance of Key Findings Relevant for the Mission of VBT: Abnormalities of the blood vessel wall is fundamental to many devastating vascular pathologies. Insights from our research will help design novel therapeutic strategies for these diseases. 4. Publications: (Publications July 1, 2013– June 30, 2014) Kim, J, Kang, Y, Kojima, Y, Lighthouse, JK, Hu, X, Aldred, MA, McLean, DL, Park, H, Comhair, SA, Greif,

DM, Erzurum, SC, Chun, HJ. (2013). A novel endothelial apelin-FGF link mediated by microRNAs 424 and 503 is disrupted in pulmonary arterial hypertension. Nature Medicine, 19:74.

Seidelmann, SB, Lighthouse, JK, Greif, DM*. (2014). Development and pathologies of the arterial wall. Cellular and Molecular Life Sciences, 71:1977. (*Corresponding author).

Sheikh, AQ, Lighthouse, JK, Greif, DM*. (2014). Recapitulation of developing artery muscularization in pulmonary hypertension. Cell Reports, 6:809. (*Corresponding author).

Greif, DM*, Eichmann, A*. (2014). Brain vessels squeezed to death. Nature, 508:50. (*Corresponding authors).

Misra, A, Sheikh, AQ, Ding, L, Kumar, A, Hinton, RB, Smoot, L, Kaplan, P, Urban, Z, Tellides G, Greif, DM*. Aortic smooth muscle in development and disease: role of integrin β3 in hypermuscularization. Manuscript submitted. (*Corresponding author).


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Jaime Grutzendler, MD Associate professor of Neurology and of Neurobiology; Director Yale Center for Experimental Neuroimaging (YCEN) 1. Overall Goal(s) of the Research Program of the Laboratory: The overall interest of our laboratory centers around cells in the so called "neuro-glio-vascular unit" (neurons, endothelium, astrocytes, pericytes, microglia, oligodendrocytes and NG2 cells). Our goal is to learn about the dynamic properties of these cells in vivo and how cell-cell interactions are disrupted in brain injury, vascular and neurodegenerative pathologies. 2. Specific Research Accomplishments in the last 12 months: We continue to make progress on the understanding and potential translation of our recent discovery of a novel mechanism of microvascular recanalization independent of the fibrinolytic system that we have named angiophagy. We have also developed novel methodologies for imaging myelinated axons in the central and peripheral nervous systems using a label-free method that we call SCoRe ( spectral confocal reflectance microscopy). We are currently incorporating this method to our in vivo imaging studies of brain development, pathology and aging. 3. Significance of Key Findings Relevant for the Mission of VBT: We recently conducted the First longitudinal in vivo imaging study of the postnatal cerebral microvasculature demonstrating that the final patterning occurs by a strategy of refinement involving sprouting and pruning (Harb et al, J Cereb Blood Flow Metab, 2013). Discovered the existence of a critical period for postnatal cerebral microvascular remodeling during which disruption of neural activity leads to permanent reductions in microvascular density (Whiteus et al, Nature, 2014). This finding has important implications potentially linking perinatal homeostatic disruptions with adult life microvascular deficits that could explain brain dysfunction and age-related pathologies. We also published a follow up paper (Grutzendler et al; Sci Transl Med. 2014) focusing on various novel aspects of the angiophagy mechanisms of microvascular recanalization that we discovered. We demonstrate that this mechanism is ubiquitous of all vascular beds, but in specific places like the lung and kidney it has unique features as emboli are able to cross the endothelium as well as the adjacent epithelium, constituting potential routes of circulating debris elimination. We also show evidence suggesting that angiophagy occurs in the human retina in vivo. 4. Publications (Publications July 1, 2013– June 30, 2014) J. Grutzendler. Angiophagy. Mechanism of Microvascular Recanalization Independent of the Fibrinolytic

System. Stroke. 2013: 44:S84-S86 C. Whiteus, C. Freitas, J. Grutzendler. Perturbed neural activity disrupts cerebral angiogenesis during a

postnatal critical period. Nature. 2014 Jan 16. Epub 2013 Dec 4. J. Grutzendler, S. Murikinati, B. Hiner, C. Lam , T. Yoo, L. Ji,B, Hafler, R. Adelman, P. Yuan, S. Gupta, G.

Rodriguez. Angiophagy prevents early embolus washout but leads to microvascular recanalization through embolus extravasation. Science Translational Medicine. 2014 Mar 5;6 (226).

A. Schain, R. Hill, J. Grutzendler. Label-free in vivo imaging of myelinated axons in health and disease with spectral confocal reflectance microscopy. Nature Medicine. 2014 Apr;20(4):443-9.

Zhang X, Tian Y, Yuan P, Li Y, Yaseen MA, Grutzendler J, Moore A, Ran C. A bifunctional curcumin analogue for two-photon imaging and inhibiting crosslinking of amyloid beta in Alzheimer's disease. Chem Commun (Camb). 2014 Aug 19.

R. Hill and J. Grutzendler. In vivo imaging of oligodendrocytes using sulforhodamine 101. Nature Methods (In Press).

R. Hill , K Patel, J. Grutzendler, A. Nishiyama. Oligodendrocyte fate is modulated by the neural microenvironment during a precise temporal window after progenitor division. Nature Neuroscience (In Press)


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Karen Hirschi, Ph.D. Professor of Medicine (Cardiology) 1. Overall Goal(s) of the Research Program of the Laboratory: A primary interest of our laboratory is to understand, at the cellular and molecular level, the events leading to blood vessel formation. We are specifically interested in the regulation of vascular cell commitment, differentiation and cell cycle progression, and defining signaling pathways that modulate these processes. We also aim to understand how some vascular cells acquire specialized functions, such as the generation and/or maintenance of multi-lineage stem/progenitor cells. 2. Publications: (Publications July 1, 2013– June 30, 2014) Hirschi KK, Li S and Roy K: Induced pluripotent stem cells for regenerative medicine. Annu Rev Biomed

Eng. 2014 Jul 11;16 :277-94. Epub 2014 May 29. PMID: 24905879 Marcelo KL, Sills TM, Coskun S, Vasavada H, Sanglikar S, Goldie LC and Hirschi KK: Hemogenic

endothelial cell specification requires c-Kit, Notch signaling, and p27-mediated cell-cycle control. Dev Cell. 2013 Dec 9;27 (5) :504-15. PMID: 24331925


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Jay D. Humphrey, Ph.D. John C. Malone Professor, Department of Biomedical Engineering

1. Overall Goal(s) of the Research Program of the Laboratory: The primary goal of my laboratory is to understand better and mathematically model the roles of mechanobiological mechanisms in tissue-level homeostasis, adaptation, and pathogenesis. Specific interests include arterial changes in response to hypertension, altered flow, and aging, the design and assessment of tissue engineered vascular grafts, venous changes when used as arterial grafts, and the progression of large artery diseases such as aortic and cerebral aneurysms. We are also interested in exploiting genetically modified mice to elucidate individual contributions of extracellular matrix constituents (e.g., fibrillin-1, fibulin-5, collagens III and V, and GAGs) to tissue level arterial structure and function. 2. Specific Research Accomplishments in the last 12 months: One of the main findings is the potential adverse role of accumulated GAGs within the thoracic aorta, which may be causative in aortic dissection. Another is the development of a computational model that describes well the in vivo development of a neovessel from an implanted polymeric tissue engineered vascular graft. 3. Significance of Key Findings Relevant for the Mission of VBT: Our ability to assess biomechanically the multiaxial mechanical properties of mouse arteries and veins supported multiple projects by investigators within the VBT and has enabled the initiation of new collaborative projects. 4. Publications: (Publications July 1, 2013– June 30, 2014) Valentin A, Humphrey JD, Holzapfel GA (2013) A 3D finite element based constrained mixture framework

for aortic growth and remodeling. Int J Num Meth Biomed Engr 29: 822-849. Lee YU, Naito Y, Kurobe H, Breuer CK, Humphrey JD (2013) Biaxial mechanical properties of the inferior

vena cava in C57BL/6 and C-17 SCID/bg mice. J Biomech 46: 2277-2282. Roccabianca S, Figueroa CA, Tellides G, Humphrey JD (2014) Quantification of regional differences in

aortic stiffness in the aging human aorta. J Biomech Behavior Biomed Matl 29: 618-634. Roccabianca S, Ateshian G, Humphrey JD (2014) Biomechanical roles of medial pooling of

glycosaminoglycans in thoracic aortic dissection. Biomech Model Mechanobiol (ePub ahead of press). Naito Y, Lee YU, Yi T, Church SN, Solomon D, Humphrey JD, Shinoka T, Breuer CK (2014) Beyond burst

pressure: Initial evaluation of the natural history of the biaxial mechanical properties of tissue engineered vascular grafts in the venous circulation using a murine model. Tiss Engr Part A 20: 346-355.

Bellini C, Ferruzzi J, Roccabianca S, DiMartino E, Humphrey JD (2014) A microstructurally-motivated model of arterial wall mechanics with mechanobiological implications. Annl Biomed Engr 42: 488-502.

Miller KS, Lee YU, Naito Y, Breuer CK, Humphrey JD (2014) Computational model of the in vivo development of a tissue engineered vein from an implanted polymeric construct. J Biomech 47: 2080-2087.

Li W, Li Q, Jiao Y, Qin L, Ali R, Zhou J, Ferruzzi J, Kim RW, Geirsson A, Dietz HC, Offermanns S, Humphrey JD, Tellides G (2014) Tgfbr2 disruption in postnatal smooth muscle impairs aortic wall homeostasis. J Clin Invest 124: 755-767.

Bersi MR, Ferruzzi J, Eberth JF, Gleason RL, Humphrey JD (2014) Consistent biomechanical phenotyping of common carotid arteries from seven different genetic, pharmacological, and surgical mouse models. Annl Biomed Engr 42: 1207-1223.

Genovese K, Casaletto L, Humphrey JD, Lu J (2014) Digital image correlation based point-wise characterization of heterogeneous material properties of gallbladder in vitro. Proceed R Soc Lond 470: 20140152


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Udelsman BV, Khosravi R, Miller KS, Dean EW, Bersi MR, Rocco K, Yi T, Humphrey JD, Breuer CK

(2014) Characterization of evolving biomechanical properties of tissue engineered vascular grafts in the arterial circulation. J Biomech 47: 2070-2079.

Humphrey JD, Milewicz DM, Tellides G, Schwartz MA (2014) Dysfunctional mechanosensing in aneurysms. Science 344: 477-479.

Bai Y, Lee P-F, Humphrey JD, Yeh AT (2014) Intravital characterization of engineered tissues by multimodal optical imaging and biaxial mechanical testing. Annl Biomed Engr 42: 1791-1805.

Roccabianca S, Bellini C, Humphrey JD (2014) Computational modeling suggests good, bad, and ugly roles of glycosaminoglycans in arterial mechanics and mechanobiology. J Roy Soc Interface (ePub ahead of print)


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Name John Hwa, M.D., Ph.D. Title & Department Associate Professor of Medicine (Cardiology)

1. Overall Goal(s) of the Research Program of the Laboratory: The major focus of our laboratory is to decipher the mechanisms leading to diabetic platelet dysfunction. Our overall hypothesis is; “aldose reductase is a major transducer of the hyperglycemic response, resulting in mitochondrial dysfunction and damage and a prothrombotic state”. To test our hypotheses we study human patients and human tissues (clinical, pathophysiology, pharmacology, cell biology, molecular biology, and bioinformatics) in combination with animal studies. Our ultimate goals are to identify patients at increased risk for atherothrombosis, and to develop novel therapies targeting atherothrombosis.

2. Specific Research Accomplishments in the last 12 months: Platelet abnormalities are well-recognized complications of diabetes mellitus (DM). Mitochondria play a central role in platelet metabolism and activation. Mitochondrial dysfunction is evident in DM. The molecular pathway for hyperglycemia-induced platelet mitochondrial dysfunction in DM platelets is unknown. Using both human and humanized mouse models, we have demonstrated that hyperglycemia-induced aldose reductase (AR) activation, and subsequent reactive oxygen species (ROS) production, leads to increased p53 phosphorylation (Ser15), which promotes mitochondrial dysfunction, damage and rupture by sequestration of the anti-apoptotic protein Bcl-xL. In a glucose dose dependent manner, severe mitochondrial damage leads to loss of mitochondrial membrane potential and platelet apoptosis (cytochrome c release, caspase 3 activation and phosphatidylserine exposure). Although platelet hyperactivation, mitochondrial dysfunction, AR activation, ROS production and p53 phosphorylation are all induced by hyperglycemia, we demonstrate that platelet apoptosis and hyperactivation are two distinct states, dependent upon the severity of the hyperglycemia and mitochondrial damage. AR contributes to diabetes-mediated mitochondrial dysfunction and damage through the activation of p53. The degree of mitochondrial dysfunction and damage determines whether hyperactivity (mild damage) or apoptosis (severe damage) will ensue. Further studies are underway assessing how the hyperglycemia induced mitochondrial damage affects platelet metabolism, how autophagy may serve a protective role, how the subsequent production of ROS may regulate VWF and the pharmacogenetics of these pathways. These signaling components provide novel therapeutic targets for DM thrombotic complications.

3. Significance of Key Findings Relevant for the Mission of VBT: Identification of the role played by aldose reductase in platelet mitochondrial damage Identification of potential novel therapeutic targets

4. Publications: (Publications July 1, 2013– June 30, 2014) Chakraborty R., Pydi SP., Gleim S., Bhullar RP., Hwa J., Dakshinamurti D., Chelikani P. New insights into

structural determinants for prostanoid receptor-G-protein specificity. Mol Cell Biol 2013 33(2):184-193 Lemaire M., Fremeaux-Bacchi V., Schaeffer F., Choi M., Tang W.H., Taque S., Nobili F., Martinez F., Ji W.,

Overton J.D., Mane S.M., Fakhouri F., Adra A.L., Deschenes G., Baudoin V., Llanas B., Collard L., Majid

M.A., Simkova E., Nuernberg P., Rioux-Leclerc N., Moeckel G.W., Gubler M.C., Hwa J., Loirat C., & Lifton R.P. Recessive mutations in diacylglycerol kinase epsilon cause atypical hemolytic-uremic syndrome. Nature Genetics 2013 45(5):531-6

Kang Y., Anderson J., Kim J., Gleim S., Kundu R., McLean D., Park H., Hwa J., Quertermous T., Chun H.J. Apelin-APJ signaling is a critical regulator of endothelial MEF2 activation in murine cardiovascular development. Circ Research 2013 113(1):22-31

Gleim S., Stitham J., Tang W.H., Li H., Douville K., Chelikani P., Rade J.J., Martin K., Hwa J. Human thromboxane A2 receptor Genetic Variants: In silico, in vitro and “in platelet” analysis. PLoS One 2013 8(6):e67314

Liu R, Yu J, Jin Y, Tang W, Qin L, Zhang X, Tellides G, Hwa J, Martin KA. TET2 is an mTORC1-dependent master regulator of vascular smooth muscle plasticity. Circulation 2013, 128(18):2047-57


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Frey A., Ibrahim S., Gleim S., Hwa J., Smyth E.M.. Biased Suppression of Thromboxane A2 Receptor

Homodimerization and Signaling Through Disruption of a Transmembrane GxxxGxxxL Helical Interaction Motif. J Lipid Res 2013 54(6):1678-90

Partovian C., Li SX., Xu X., Lin H., Lin H., Strait K.M., Hwa J., Krumholz HM. Distinct patterns of hospital trend in niseritide use for patients with heart failure. J Am Coll Cardiol - HF 2013 1(4):318-324

Chakraborty R., Bhuller R.P., Dakshinamurti S., Hwa J., Chelikani P. Inverse agonism of SQ 29,548 and ramatroban on the thromboxane A2 receptor PLoS One 2014 9(1):e85937

Tang W.H., Stitham J., Liu R., Gleim S., Spollett G., Martin K., Hwa J. Hyperglycemia promotes platelet apoptosis through a unique pathway involving aldose reductase, ROS, p53 and Bcl-xl. Circulation 2014 129(15):1598-609

Keramati A.R., Fathzadeh M., Singh R., Choi M., Faramarzi S., Mane S., Kasaei M., Sarajzadeh-Fard K., Hwa J., Kidd K.K., Babaee Bigi M.A., Malekzadeh R., Hosseinian A., Babaie M, Lifton R.P., Mani A.. Altered function of Dyrk1B is Associated with a Highly Penetrant Form of Obesity, Atherosclerosis and their Metabolic Risk Factors. N Engl J Med 2014 370(20):1909-19

Moore J., Hwa J.. (Editors) Pharmacogenomics and pharmacogenetics: “So close and yet so far”. Thematic issue for Curr Mol Med (in press)

Stitham J., Vanichakarn P., Lee Y., Hwa J. Cardiovascular pharmcogenetics of antithrombotic agents and NSAIDs. Curr Mol Med (in press)

Vanichakarn P., Hwa J., Stitham J. Cardiovascular pharmacogenetics of antihypertensive and lipid lowering agents. Curr Mol Med (in press)


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Yasuko Iwakiri, Ph.D Associate Professor of Medicine (Digestive Diseases) 1. Overall Goals of the Research Program of the Laboratory: The main areas of my research include vascular remodeling in liver diseases, liver fibrosis and liver regeneration. We are especially interested in how hemodynamic changes are associated with liver fibrosis and liver regeneration and how endothelial cell dysfunction leads to liver fibrosis. The goals are to understand the basic mechanisms of flow-induced changes in the liver and to apply this knowledge to develop therapeutic strategies for liver fibrosis/cirrhosis and portal hypertension. 2. Specific Research Accomplishments in the last 12 months: We have demonstrated Nogo-B promotes liver fibrosis and portal hypertension. We have also found the mechanisms of arterial thinning in portal hypertension with liver cirrhosis. 3. Significance of Key Findings Relevant for the Mission of VBT: Research conducted by our laboratory focuses on development of therapeutic strategies that modify vasculature/endothelial cell function in a manner to prevent liver fibrosis in portal hypertension. Publications (from July 1, 2013 to June 30, 2014): Tashiro, K., Satoh, A., Utsumi, T., Chung, C., and Iwakiri, Y. Absence of Nogo-B (Reticulon 4B) facilitates

hepatic stellate cell apoptosis and diminishes hepatic fibrosis in mice. Am J Pathology. 2013;182(3):786-95.

Gao, L., Utsumi, U., Tashiro, K., Liu. B., Zhang, D., Swenson, E.S., and Iwakiri, Y. Reticulon 4B (Nogo-B) facilitates hepatocyte proliferation and liver regeneration. Hepatology. 2013; 57(5):1992-2003.

Bryniarski, K., Ptak, W., Jayakumar, A., Tuschl, T., Hafner, M., Püllmann, K., Caplan, M., Chairoungdua, A., Lu, J., Adams, B., Sikora, E., Nazimek, K., Marques, S., Kleinstein, S.H., Sangwung, P., Iwakiri, Y., Delgato, E., Redegeld, F., Wojcikowski, J., Daniel, A.W., Kormelink, T.G., and Askenase, P.W. Antibody light chain coated antigen specific exosomes deliver suppressor T cell-derived miRNA-150 to inhibit effector T cells. J Allergy Clinical Immunology. 2013; 132(1):170-181. PMID: 23727037.

Gattu, A., Iwakiri, Y., Chung, C. et al. Determination of Mesenchymal Stem Cell Fate by Pigment Epithelium-Derived Factor (PEDF) Results in Increased Adiposity & Reduced Bone Mineral Content. FASEB J. 2013;27(11):4384-94.

DiLorenzo, A., Lin, M., Murata, T., Landskroner-Eiger, S., Schleicher, M., Kothiya, M., Iwakiri, Y., Yu, J., Huang, P., and Sessa, W. eNOS derived nitric oxide regulates endothelial barrier function via VE cadherin and Rho GTPases. Journal of Cell Science. 2013;126(Pt 24):5541-52.

Chung, C. and Iwakiri, Y. The lymphatic vascular system in liver diseases: its role in ascites formation. Clinical and Molecular Hepatology. 2013; 19(2):99-104.

Gattu, A.K., Birkenfeld, A.L., Protiva, P., Church, C., Jay, S., Saltzman, M., Doll, J., Cornwell, M., Crawford, S.E., Iwakiri, Y., Samuel, V.T., and Chung, C. Pigment Epithelium-Derived Factor (PEDF) Blunts IL-1β-Mediated Inflammation in Hepatocytes to Maintain Metabolic Homeostasis. Endocrinology. 2014; 155(4): 1373-85.

Jozsef, L., Tashiro, K., Kuo, A., Park, E., Skoura, A., Albinsson, S., Rivera-Molina, F., Harrison, K.D., Iwakiri, Y., Toomre, D., and Sessa, W.C. Reticulon 4 is necessary for ER tabulation, STIM1-Orai1 coupling and store-operated calcium entry. J Biol. Chem. 2014; 289(13): 9380-95.

Iwakiri, Y. Pathophysiology of portal hypertension. Clinics Digestive Diseases. 2014; 19(2):99-104. Iwakiri, Y., Shah, V., and Rockey, D. Vascular Pathology in Chronic Liver Disease and Cirrhosis – Current

Status and Future Directions. J of Hepatology. 2014 (in press)


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Suk-Won Jin, Ph.D. Associate Professor (Adjunct) of Medicine (Cardiology) 1. Overall Goal(s) of the Research Program of the Laboratory: My laboratory is interested in understanding molecular and cellular mechanisms that regulate developmental and pathological angiogenesis, using zebrafish as a model system. Currently our major efforts are directed at analyzing the function of Bone Morphogenetic Protein (BMP) signaling during vascular patterning, investigating interaction between endothelial and hematopoietic lineages during development, and analyzing molecular and cellular defects of vascular specific mutations. 2. Specific Research Accomplishments in the last 12 months: We have reported that BMP2 signaling negatively modulates the differentiation of lymphatic endothelial cells while promoting venous angiogenesis (Dunworth et al, Circ. Res, 2014). In addition, we have identified Apelin-APJ signaling as one of the major inputs that modulate lymphatic development. 3. Significance of Key Findings Relevant for the Mission of VBT: Kim JD, Kang Y, Kim J, Papangeli I, Kang H, Wu J, Park H, Nadelmann E, Rockson SG, Chun HJ and Jin

SW: Essential role of Apelin signaling during lymphatic development in zebrafish. Arterioscler Thromb Vasc Biol. 2014 Feb;34 (2) :338-45. Epub 2013 Dec 5. PMID: 24311379

Dunworth WP, Cardona-Costa J, Bozkulak EC, Kim JD, Meadows S, Fischer JC, Wang Y, Cleaver O, Qyang Y, Ober EA and Jin SW: Bone morphogenetic protein 2 signaling negatively modulates lymphatic development in vertebrate embryos. Circ Res. 2014 Jan 3;114 (1) :56-66. Epub 2013 Oct 11. PMID: 24122719

4. Publications: (Publications July 1, 2013– June 30, 2014) Nie L, Guo X, Esmailzadeh L, Zhang J, Asadi A, Collinge M, Li X, Kim JD, Woolls M, Jin SW, Dubrac A,

Eichmann A, Simons M, Bender JR and Sadeghi MM: Transmembrane protein ESDN promotes endothelial VEGF signaling and regulates angiogenesis. J Clin Invest. 2013 Dec 2;123 (12) :5082-97. Epub 2013 Nov 1. PMID: 24177422

Kim JD, Lee HW and Jin SW: Diversity is in my veins: role of bone morphogenetic protein signaling during venous morphogenesis in zebrafish illustrates the heterogeneity within endothelial cells. Arterioscler Thromb Vasc Biol. 2014 Sep;34 (9) :1838-45. Epub 2014 Jul 24. PMID: 25060789

Kim JD and Jin SW: A tale of two models: mouse and zebrafish as complementary models for lymphatic studies. Mol Cells. 2014 Jul 31;37 (7) :503-10. Epub 2014 May 23. PMID: 24854860

Sakurai T, Woolls MJ, Jin SW, Murakami M and Simons M: Inter-cellular exchange of cellular components via VE-cadherin-dependent trans-endocytosis. PLoS One. 2014;9 (6) :e90736. Epub 2014 Mar 6. PMID: 24603875


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Martin S. Kluger, Ph.D. Research Scientist, Department of Immunobiology 1. Overall Goals of the Research Program of the Laboratory: To understand how inflammatory cytokines produce capillary leak by targeting endothelial cell (EC) intercellular junctions. In particular, we study tumor necrosis factor (TNF) signaling pathways that affect the tight junctional molecule claudin-5 and claudin-5-associated proteins. These studies relate to the uncontrolled paracellular transit of fluid and proteins in capillary beds, which is critical to the pathogenesis of blood sepsis and systemic inflammatory response syndrome (SIRS). 2. Specific Research Accomplishments in the Last 12 Months:

We made novel observations regarding NF-B and ROCK-dependent mechanisms triggered by cytokines (TNF and IL-1that alter tight junction-dependent permeability barriers formed by human microvascular endothelial cells. These studies were possible using the model we developed for this purpose (published as “Claudin-5 Controls Intercellular Barriers of Human Dermal Microvascular but Not Human Umbilical Vein Endothelial Cells. Arterioscler Thromb Vasc Biol. 2013. 33:489-500).

New lab recruit Richard Pierce, MD is being trained by Dr. Kluger to perform translational research aimed at understanding the biological basis for microvascular leak affecting pediatric patients at Yale/New Haven Hospital. Dr. Pierce’s research effort is funded by a position in the Yale Pediatric Basic Science Training Program and fostered by his clinical mentor John S. Giuliano Jr. MD, Assistant Professor of Pediatrics (Critical Care). Collaborative studies on this and other directions continue with Dr. Jordan Pober and in projects performed by lab member Paul Clark, PhD.

Dr. Kluger completed a 3rd successful year co-directing (with Dr. Jun Yu) the well-attended VBT Research-In-Progress Series, a seminar series that promotes research interactions among different VBT laboratories housed in the Amistad Research building. This year graduate students and post-docs presented research accomplishments from the Fernandez, Giordano, Kyriakides, Min, Niklason, Pober, Saltzman, Sessa, Suarez, Tellides, Wu and Yu labs.

3. Significance of Key Findings Relevant for the Mission of VBT: We report for the first time that cytokine-induced hyperpermeability takes place in two discrete phases, and that prevention of capillary leak may require targeting more than one pathway. The conclusions drawn from these studies suggest strategies for preventing disruption of the TJs specifically found in the capillary (as distinguished from the post-capillary) segments of failing organs in SIRS or septic patients. 4. Publications: Clark, P.R., Pober, J.S., and Kluger, MS TNF induces three sequential changes in human dermal

microvascular endothelial cell barriers: an in vitro model of inflammatory capillary leak. Angiogenesis, 2014. 17:311.

Clark, P. R., Kim, R.K., Pober, J.S. and Kluger, MS TNF Disrupts Claudin-5 Endothelial Tight Junction Barriers in Two Distinct NF-B-Dependent Phases. Submittted for publication.


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Diane Krause MD, PhD

Professor of Laboratory Medicine

1. Overall Goal(s) of the Research Program of the Laboratory: The overall goals of my research are to characterize bone marrow derived stem and progenitor cells, and to define the molecular mechanisms (signal transduction, biomechanical, and epigenetic) that regulate the self-renewal and differentiation of these cells. Our recent emphasis has been on megakaryocyte development, megakaryoblastic leukemia, as well as platelet formation. We have a renewed interest now on platelet function. We are studying the roles of G-proteins, the SRF signal transduction pathway, and RNA binding proteins in order to better understand and treat hematopoietic diseases including myelodysplasia, myeloproliferative disease and leukemia, as well as vascular diseases related to thrombus formation and acute infarction. Projects include work with embryonic stem cells as well as hematopoietic stem cells from mice and humans. Our work provides insights not only into normal blood cell development, but also to the pathogenesis of benign and malignant hematological diseases.

2. Specific Research Accomplishments in the last 12 months Over the past year, we have published 4 papers, and acquired a he amount of exciting new data that will be published over the coming year. Dr. Yeun-hee Kim published data from her in depth study on the molecular evaluation of primary acute megakaryocytic leukemia (AMKL)> After performing genome capture - deep seq of disease and remission bone marrow cells, she discovered several mutated genes, and confirmed each one by DOP-PCR. The most promising candidate somatic mutation to play a role in cellular transformation was MMP8, a secreted matric metalloproteinase that is critical for cell interactions with the microenvironment. This was published in the excellent peer review journal Leukemia. Another very exciting paper resulted from collaboration between my laboratory and that of Dr. Shangqin Guo at Yale. We published in Cell that then hematopoietic progenitors cells are induced to undergo reprogramming to become induced pluripotent stem cells, the process I not stochastic. Rather, there is a small population of rapidly proliferating cells that are the source of the subsequent iPS cells. How a raid cell cycle causes a cell to be more prone to reprogramming is currently under investigation.

3. Significance of Key Findings Relevant for the Mission of VBT Studies on megakaryocytes and platelets are highly relevant to the mission of VBT because platelets play a key role in atherosclerosis and inflammation and perhaps in development of lymphatic endothelial lining cells as well. The means by which they are produced, and the transcriptional regulation of the genes expressed by platelets are highly relevant to vascular biology. In addition, the SRF/MKL1 pathways on which we focus is highly relevant to the dysfunction of smooth muscle cells in vascular diseases. 4. Publications:

Krause DS, Crispino J. Molecular Pathways: Induction of polyploidy as a novel differentiation therapy for leukemia. Clinical Cancer Research. 22:6084-8, 2013.

Zhang PX, Murray TS, Villella VR, Ferrari E, Esposito S, D'Souza A, Raia V, Maiuri L, Krause DS, Egan ME, Bruscia EM. Reduced caveolin-1 promotes hyperinflammation due to abnormal heme oxygenase-1 localization in lipopolysaccharide-challenged macrophages with dysfunctional cystic fibrosis transmembrane conductance regulator. J Immunol. 190:5196-206, 2013.

Kim Y, Schulz VP, Satake N, Gruber TA, Teixeira A, Halene S, Gallagher PG, Krause DS. Whole exome sequencing identifies a novel somatic mutation in MMP8 associated with a t(1;22)-acute megakaryoblastic leukemia. Leukemia. 28:945-8, 2014.


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Themis R. Kyriakides Ph.D. Associate Professor, Pathology and Biomedical Engineering 1. Overall Goal (s) of the Research Program of the Laboratory The main area of my research is the elucidation of the molecular events that dictate the course of healing and especially inflammation and angiogenesis following ischemia, injury and the implantation of biomaterials and scaffolds for tissue engineering applications. Our primary research focus is on two molecules, MCP-1 and TSP-2 that we have shown to be critical to various aspects of these processes. In a new research effort we are exploring the inter-relationship between TSP2 and the Akt/eNOS signaling axis. In addition, through the process of molecular dissection of cell-matrix interactions, we aim to incorporate rational design in the development of bioengineering applications such as tissue-engineered vascular grafts. Finally, we aim to define role of TSP2 in the maintenance and repair of the blood brain barrier in the context of brain-biomaterial interactions. 2. Specific Research Accomplishments in the last 12 months (7/1/13-6/30/14) We have continued our investigation of the participation of TSP-2 in angiogenesis and arteriogenesis. Furthermore, we have explored the participation of TSP2/AKt/eNOS signaling axis in endothelial and fibroblast function. We have generated double eNOS/TSP2-null mice and Akt1/TSP2-null mice and discovered that the absence of TSP2 ameliorates the phenotypes of the eNOS-null and Akt1-null mice. More recently, we have shown in in vivo and vitro studies that nitric oxide and hypoxia suppress TSP2 expression at the transcriptional level. In our biomedical engineering-related research we have discovered that MCP-1 contributes to biomaterial-induced inflammation and subsequent fibrosis by inducing TNF-α and the canonical NFyb pathway. Within the VBT program we have continued our collaborations with the following investigators: Sessa, Simons, Schwartz, Giordano, Saltzman, Niklason, and Tellides. 3. Significance of Key Findings Relevant for the Mission of VBT Studies in angiogenesis, arteriogenesis, and engineering of vascular grafts are central to the mission of the VBT. In addition, our studies investigating the link between TSP2, Akt1, and eNOS are of importance to many processes in vascular biology. 4. Publications (2013-2014) Sawyer A.J., Tian W., Saucier-Sawyer J.K., Rizk P., Saltzman W.M., Bellamkonda R., Kyriakides T.R.

(2014) The effect of inflammatory cell-derived MCP-1 loss on neuronal survival during chronic

neuroinflammation. Biomaterials 35: 6698-706. Padmanabhan J., Kinser E.R., Stalter M.A., Duncan-Lewis C., Balestrini J.L., Sawyer A.J., Schroers J,

Kyriakides T.R. (2014) Engineering Cellular Response Using Nanopatterned Bulk Metallic Glass. ACS Nano 8:4366-75.

Andrew J. Sawyer, Themis R. Kyriakides. (2013) Nanoparticle-based evaluation of blood brain-barrier leakage during the foreign body response. J. Neural Eng. 10: 016013.

Calabro N, Kristofik N, Kyriakides TR. (2014) Thrombospondin-2 and extracellular matrix assembly. Biochem. Biophys. Acta 1840:2396-402

Morris A, Kyriakides TR. Matricellular proteins and biomaterials. Matrix Biology [Epub ahead of print] Kobsa S, Kristofik NJ, Sawyer AJ, Bothwell AL, Kyriakides TR*, Saltzman WM* (2013) An electrospun

scaffold integrating nucleic acid delivery for treatment of full-thickness wounds. Biomaterials 34:3891-901.

A. J. Sawyer, D. Wesolowski, N. Gandotra, A. Stojadinovic, M. Izadjoo, S. Altman, Kyriakides TR (2013) A peptide-morpholino oligomer conjugate targeting Staphylococcus aureus gyrA mRNA improves healing in an infected mouse cutaneous wound model. Int. J. Pharmaceutics 453(2):651-5.

Ju R, Zhuang ZW, Zhang J, Lanahan AA, Kyriakides TR, Sessa WC, Simons M. Angiopoietin-2 Secretion by Endothelial Cell Exosomes: regulation by the phosphatidylinositol 3-kinase (PI3K)/Akt/ endothelial nitric oxide synthase (eNOS) and syndecan4/syntetin pathways. (2014) J. Biol. Chem. 289:510-9


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Swartzlander MD, Lynn AD, Blakney AK, Kyriakides TR, and Bryant SJ. (2013) Understanding the Host

Response to Cell-Laden Poly(ethylene glycol)-based Hydrogels. Biomaterials 34:952-64. Lee MY, Luciano AK, Ackah D, Rodriguez-Vita K, Bancroft TA, Eichmann A, Simons M, Kyriakides TR,

Morales-Ruiz M, Sessa WC. Endothelial Akt1 mediates angiogenesis by phosphorylating multiple angiogenic substrates. PNAS 111:12865-70.

Moore LB, Sawyer AJ, Charokopos A, Skokos Ea, Kyriakides TR. Loss of MCP-1 alters macrophage

polarization and reduces NFκB activation in the foreign body response. Acta Biomaterialia (accepted).


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Patty J. Lee, M.D. Associate Professor of Medicine (Pulmonary) 1. Overall Goal(s) of the Research Program of the Laboratory: My laboratory investigates mechanisms of lung injury and cytoprotection during oxidant stress. Specifically, we have focused on the lung endothelium as a central mediator of lung injury and repair responses. We identified the importance of the stress-response protein heme oxygenase-1 (HO-1) and its gaseous reaction product, carbon monoxide (CO), in resisting oxidant-induced endothelial cell death via mitochondrial pathways. We found that a family of signaling molecules, mitogen-activated protein kinases (MAPKs), mediates HO-1's and CO’s protective effects as well as optimal IL-13-induced lung inflammation / remodeling and, more recently, critical innate immune responses. The innate immune system consists of pattern-recognition receptors called toll-like receptors (TLRs), of which TLR4 is the LPS-responsive receptor. We discovered that TLR4 is required for lung structural cell survival in aging and oxidant challenges. These studies represent important paradigm shifts in our understanding of TLRs and lung biology and are now the basis of translational studies in people with acute lung injury and age-related chronic lung disease, such as chronic obstructive pulmonary disease (COPD). In the process of our investigations, we were the first to demonstrate the utility of intranasal, lung-targeted and endothelial-targeted silencing RNA (siRNA) constructs in vivo. In parallel, we have also generated endothelial-targeted transgenic and knockout mouse models to specifically interrogate the role of the endothelium in lung disease. Our coordinated use of siRNA technology and genetic approaches in both cell and mouse models offer immense insight into disease pathogenesis and may identify novel therapeutic targets for a range of lung diseases. 2. Specific Research Accomplishments in the last 12 months: Her key accomplishments for the year include creating lung-endothelial targeting vectors that can be used in vivo, identifying novel molecules downstream of TLR signaling and showing for the first that mitochondrial turnover and fission are critical determinants of COPD as well as susceptibility to sepsis. Two of these publications received American Physiologic Society Awards for “Best New Publications” and one of the publications was named “Editor’s Pick for top new research publications, Am J Physiol Lung Cell Mol Biol. She also assembled a group of multi-disciplinary investigators to study COPD in the context of immunologic aging. The investigators are from Yale Pathology, Comparative Medicine, Program on Aging, Rheumatology and Infectious Diseases. Their collaborations in the past year alone have resulted in a revised P01 submission grant, which scored 24, and that will be resubmitted in Jan. 2015. Based on productive collaborations with the Aging Lung group, Drs. Bucala and Lee are co-editing a monograph, “Aging Lung: Mechanisms and Clinical Sequela” and Drs. Fragoso and Lee were invited to co-edit a COPD chapter in Hazzard’s textbook of Gerontology, 7th Edition. 3. Significance of Key Findings Relevant for the Mission of VBT: Dr. Lee’s work aligns well with the mission of VBT by introducing new vascular biology reagents, specifically in lung vasculature, and forming multi-disciplinary research groups to study novel aspects of lung biology. 4. Publications: (Publications July 1, 2014–current) Sauler M, Leng L, Trentalange M, Haslip M, Shan P, Piecychna M, Andrews N, Allore H, Fried T, Bucala R

and Lee PJ. Macrophage migration inhibitory factor deficiency in chronic obstructive pulmonary disease, Am J Physiol Lung Cell Mol Physiol, ePub Jan 17, 2014.

Mannam P, Shinn A, Srivastava A, Neamu R, Walker W, Bohanon, M, Merkel J, Kang M-J, Dela Cruz C, Ahasic A, Pisani M, Trentalange M, West A.P, Shadel G, Shen J, Elias J and Lee PJ. MKK3

regulates mitochondrial biogenesis and mitophagy in sepsis-induced lung injury, Am J Physiol Lung Cell Mol Physiol, 2014, ePub Jan 31, 2014.


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Zhang Y, Sauler M, Shinn, AS, Gong, H, Haslip M, Shan P, Mannam P and Lee PJ. Endothelial PINK1

mediates the protective effects of NLRP3 deficiency during lethal oxidant injury, J Immunol, 2014, ePub April 28, 2014.

Zhang X, Shan P, Homer R, Zhang Y, Petrache I, Mannam P and Lee PJ. Cathepsin E causes pulmonary emphysema via mitochondrial fission, Am J Pathol, 2014, In Press.

Attia, EF, Akgun, KM, Wongtrakool C, Goetz MB, Rodriguez-Barradas MC, Rimland D, Brown, ST, Hoo GS, Kim J, Lee, PJ, Schnapp LM, Sharafkhaneh A, Justice A and Crothers K. Increased risk of radiographic emphysema in HIV is associated with elevated soluble CD14 and nadir CD14, Chest, 2014, ePub Jul 31, 2014.

Wegiel, B, Chin BY, Harris C, Mannam P, Kaczmarek E, Lee PJ, Zuckerbraun B, Flavell RA, Soares M and Otterbein, LE. Bacterial sensing by macrophages through carbon-monoxide-dependent inflammasome activation, J Clin Invest, In Press.


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Joseph A. Madri, M.D., Ph.D. Professor, Dept. of Pathology and Molecular, Cellular and Developmental Biology 1. Overall Goal(s) of the Research Program of the Laboratory: This past year we have been investigating specific mouse strains, which mimic the wide range of responses to hypoxia observed in the human very low birth weight premature infant population. We have found these strains to exhibit significant differences in selected signaling nodes (GSK-3β and HIF-1α), growth factors and neurotrophins that have been shown to be involved in the responses to hypoxia. In a recent study we have found that a decreased expression of a particular transcription factor (Sox10) correlates with a poor response to chronic hypoxia in mouse pups, resulting in multiple neurodevelopmental deficits. Sox10 regulates oligodendrocytogenesis, differentiation and myelination, known modulators of synaptic transmission and neuronal impulse. Currently, we have identified a small molecule that induces Sox10 and improves cognitive behavior in mice following hypoxic insult. These studies should lead to a more complete understanding of the proteins and pathways involved and provide us with needed insights for rational drug design. 2. Specific Accomplishments in the last 12 months: We have also been investigating role of Hippo pathway signaling components (YAP/TAZ) via endothelial cell adhesion molecules (CD44, CD31 and VE-cadherin) in brain microvascular endothelial cell proliferation and apoptosis. We found that endothelial cells lacking CD44 and CD31 escaped from contact inhibition and exhibited abnormal proliferation and apoptosis and exhibited increased expression of Survivin. 3. Publications: Li, Q, Canosa S, Michaud, M., Flynn, K., Krauthammer, M., J.A. Madri. Modeling the Neurovascular

Niche: Unbiased Transcriptome Analysis of the Murine Subventricular Zone in Response to Hypoxic Insult, PLOS One, 8(10): e76265. doi:10.1371/journal.pone.0076265,2013. http://dx.plos.org/10.1371/journal.pone.0076265

Tsuneki M, Madri JA., CD44 regulation of endothelial cell proliferation and apoptosis via modulation of CD31 and VE-cadherin expression. J Biol Chem. 2014 Feb 28;289(9):5357-70. doi: 10.1074/jbc.M113.529313. Epub 2014 Jan 14.


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Arya Mani, M.D. Associate Professor of Medicine (Cardiology) and of Genetics; Director Cardiovascular Genetics Program 1. Overall Goal(s) of the Research Program of the Laboratory: My laboratory’s major focus is the identification of genetic causes of major cardiovascular disorders and the elucidation of their pathophysiology. To achieve this, we have built strong ties at national and international levels with major cardiovascular centers. The goal is to identify and recruit patients and families with diverse cardiovascular disorders that have strong genetic components. Following identification and characterization of theses mutation we carry out physiological studies in mutation carriers, in vitro and in mouse models of the disease. 2. Specific Research Accomplishments in the last 12 months: My laboratory identified a nonconservative mutation in highly conserved kinase-like domain DYRK1B gene in subjects with early onset coronary artery disease and metabolic syndrome. Functional characterization of the disease gene revealed that non-mutant protein encoded by DYRK1B inhibits the SHH (sonic hedgehog) and Wnt signaling pathways and consequently enhances adipogenesis. Furthermore, DYRK1B promoted the expression of the key gluconeogenic enzyme glucose-6-phosphatase. The R102C allele showed gain-of- function activities by potentiating these effects. Further examination showed that the mutation causes insulin resistance. Investigation of the skeletal muscles led to identification of novel targets that can be targeted pharmaceutically to improve insulin sensitivity.

3. Significance of Key Findings Relevant for the Mission of VBT:

-Identified the key role of Wnt signaling in regulation of de novo lipogenesis and cholesterol synthesis -Established the role of Wnt/LRP6 in vascular smooth muscle cell differentiation and integrity of the vascular wall -Recognized the role of Dyrk1B in adipogenesis, gluconeogenesis and its altered function in development of coronary artery disease and diabetes

4. Publications: (Publications July 1, 2013– June 30, 2014) Song K, Wang S, Huang B, Luciano A, Srivastava R and Mani A: Plasma Cardiotrophin-1 Levels are

Associated With Hypertensive Heart Disease: A Meta-Analysis. J Clin Hypertens (Greenwich). 2014 Sep;16 (9) :686-92. Epub 2014 Jul 23. PMID: 25052897

Keramati AR, Fathzadeh M and Mani A: The metabolic syndrome and DYRK1B. N Engl J Med. 2014 Aug 21;371 (8) :785-6. PMID: 25140972

Keramati AR, Fathzadeh M, Go GW, Singh R, Choi M, Faramarzi S, Mane S, Kasaei M, Sarajzadeh-Fard K, Hwa J, Kidd KK, Babaee Bigi MA, Malekzadeh R, Hosseinian A, Babaei M, Lifton RP and Mani A: A form of the metabolic syndrome associated with mutations in DYRK1B. N Engl J Med. 2014 May 15;370 (20) :1909-19. PMID: 24827035

Go GW, Srivastava R, Hernandez-Ono A, Gang G, Smith SB, Booth CJ, Ginsberg HN and Mani A: The combined hyperlipidemia caused by impaired Wnt-LRP6 signaling is reversed by Wnt3a rescue. Cell Metab. 2014 Feb 4;19 (2) :209-20. PMID: 24506864


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Kathleen A. Martin, Ph.D. Associate Professor of Medicine (Cardiology); Associate Professor of Pharmacology

1. Overall Goal(s) of the Research Program of the Laboratory: The primary goals of the Martin lab are to understand the signaling mechanisms that regulate VSMC phenotype in order to suggest therapeutic strategies for intimal hyperplasia, graft arteriosclerosis, atherosclerosis, and hypertension. 2. Specific Research Accomplishments in the last 12 months: A major interest in our lab has been signaling through the mTOR pathway in vascular smooth muscle cells, since this is the pathway targeted by the highly effective stent therapeutic rapamycin. We have recently discovered that the mTOR pathway promotes VSMC differentiation through regulation of the DNA modifying enzyme TET2. We have identified TET2 as a novel master epigenetic regulator of VSMC phenotype. Notably, TET2 promotes changes in chromatin that lead to expression of prodifferentiation genes including SRF and myocardin and contractile genes such as SM-MHC and SM-alpha actin, while concomitantly inhibiting expression of de-differentiation-associated genes including KFL4. We report that TET2 promotes 5hmC accumulation in the promoters of contractile genes in VSMC, and that 5hmC is stably expressed in mature, differentiated VSMC in vivo, but drastically downregulated in cardiovascular pathologies such as intimal hyperplasia and atherosclerosis. Importantly, we found that TET2 overexpression can prevent intimal hyperplasia, while TET2 loss of function greatly exacerbates injury response. We have published this work, along with a review of epigenetic regulation of smooth muscle phenotype. In our ongoing close collaboration with Dr. John Hwa in the Department of Cardiology, we have continue to study how hyperglycemia and diabetes affects platelet function in atherothrombotic disease. Our most recent study identified aldose reductase regulation of p53 in mitochondrial damage in platelets. These studies have suggested novel mechanisms underlying platelet hyperactivity and dysfunction in diabetes. We also collaborated with Dr. Eva Rzucidlo, an academic vascular surgeon at Dartmouth Medical School, to define signaling pathways by which resveratrol, a cardioprotective compound found in red wine, promotes VSMC differentiation. 3. Significance of Key Findings Relevant for the Mission of VBT: The work in our own laboratory has identified TET2 as an exciting new epigenetic master regulator of VSMC phenotype. In contrast to myocardin, the master transcriptional coactivator in VSMC differentiation, we find that TET2 coordinately regulates differentiation and de-differentiation genes to promote a healthy smooth muscle phenotype. In collaboration with Dr. George Tellides, we have found that TET2 is repressed in human atherosclerotic samples. With Dr. Jun Yu, we have found that TET2 is repressed following vascular injury, and, most importantly, that TET2 overexpression can rescue intimal hyperplasia in vivo. These findings have therapeutic implications for multiple cardiovascular diseases involving smooth muscle. As VBT is a multi-disciplinary and highly collaborative program, we are enthusiastic about our additional VBT collaborations with the Hwa lab, in which our expertise in signal transduction has helped them in making exciting new discoveries about the effects of glucose on platelets in diabetics and normal subjects. 4. Publications: (Publications July 1, 2013– June 30, 2014) Original reports: Gleim, S, Stitham J, Tang W, Li H, Douville KL, Chelikani P, Rade J, Martin KA, Hwa J. Human

Thromboxane A2 Receptor Genetic Variants: In Silico, In Vitro and "In Platelet" Analysis. PLoS One, 2013 Jun 28;8(6):e67314. doi: 10.1371/journal.pone.0067314.

Liu R, Qin L, Jin Y, Tang W, Tellides G, Hwa J, Yu J, Martin KA. TET2 is a master regulator of smooth muscle cell plasticity, Circulation, 2013 128:2047-2057.


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Thompson AM, Martin KA, Rzucidlo EM. Resveratrol Induces Vascular Smooth Muscle Cell Differentiation

Through Stimulation of SirT1 and AMPK, PLoS One. 2014 Jan 8;9(1):e85495. doi: 10.1371/journal.pone.0085495.

Tang WH, Stitham J, Jin Y, Liu R, Lee SH, Du J, Atteya G, Gleim S, Spollet G, Martin K, Hwa J. Aldose reductase-mediated phosphorylation of p53 leads to mitochondrial dysfunction and damage in diabetic platelets. Circulation. 2014 Jan 28. [Epub ahead of print].

Review articles: Liu R., Leslie KL, Martin KA. Epigenetic regulation of smooth muscle cell plasticity. Biochim Biophys Acta - Gene Regulatory Mechanisms, Special Issue: Stress as a fundamental theme in

cell plasticity. 2014 Jun 15. pii: S1874-9399(14)00156-4. doi: 10.1016/j.bbagrm.2014.06.004. Guzman AK, Ding M, Xie Y, and Martin KA. Pharmacogenetics of Obesity. Curr Mol Med, 2014 Aug 11,

[Epub ahead of print]. Liu R, Jin Y, Tang W, Qin L, Zhang X, Tellides G, Hwa J, Yu J, Martin KA. Response to Letter Regarding

Article, "Ten-Eleven Translocation-2 (TET2) Is a Master Regulator of Smooth Muscle Cell Plasticity". Circulation. 2014 Aug 19;130(8):e72. doi: 10.1161/CIRCULATIONAHA.114.010272


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Wang Min, Ph.D. Professor with Tenure, Pathology 1. Overall Goal(s) of the Research Program of the Laboratory: Understanding of the fundamental molecular mechanisms for vasculogenesis, arteriogenesis and angiogenesis may lead to improved therapeutic strategies for treatment of vascular diseases. The goal in my lab is to dissect the signaling pathways in vasculature involved in vascular development, remodeling and repair related to human diseases such as vascular malformation, atherosclerosis, stroke, graft transplant rejection and tumor metastasis. 2. Specific Accomplishments in the 12 months: a) Awarded for Connecticut Innovations on Stem Cell Research: After four years’ efforts, I finally obtained CT Stem Cell award for Established Investigator Grant to define the in vivo function of TNFR2-Bmx signaling in cardiac progenitor and cardiac repair. This grant was based on our recent joint work with Dr. John Bradley at Cambridge, Dr. Jordan Pober at Yale (Publication #1) and more recent with Dr. Tim Kamp at University of Wisconsin.

Agency: CT Stem Cell Innovation Award (Established Investigator Grant) ID#: 14-SCB-YALE-17 (PI: Min) Title: TNFR2-Bmx signaling in cardiac stem cells and cardiac repair Direct costs per year: $150,000 Total costs for project period: $750,000. Project period: 9/1/14 – 8/30/18 b) SOCS1 in Graft arteriosclerosis models: Graft arteriosclerosis (GA) is the major cause of late allograft failure. Pathologic features include arterial intimal hyperplasia due to recruitment and proliferation of VSMC, which eventually causes luminal obstruction and allograft ischemia. We have established and characterized two mouse artery transplant models - a single minor histocompatibility antigen (male to female)-dependent aorta transplantation model and IFN-y-induced syngeneic graft transplantation model. SOCS1, a negative regulator of cytokine signaling, is highly expressed in endothelial cells (ECs). We observed dramatic but specific reduction of endothelial SOCS1 in human GA and atherosclerosis specimens, suggesting the importance of SOCS1 in maintaining normal endothelial function. SOCS1 deletion in mice resulted in basal EC dysfunction. After transplantation, SOCS1-deficient aortic grafts augmented leukocyte recruitment and neointima formation, whereas endothelial overexpression of SOCS1 diminished arterial rejection. Induction of endothelial adhesion molecules in early stages of GA was suppressed by the VESOCS1 transgene and this effect was confirmed in cultured aortic ECs. Moreover, VESOCS1 maintained better vascular function during GA progression. Mechanistically, endothelial SOCS1, by modulating both basal and cytokine-induced expression of the adhesion molecules PECAM-1, ICAM-1 and VCAM-1, restrained leukocyte adhesion and trans-endothelial migration during inflammatory cell infiltration. Our study concludes that SOCS1 prevents GA progression by preserving endothelial function and attenuating cytokine-induced adhesion molecule expression in vascular endothelium. This work was recently published in J Amer Coll Cardiol (the impact factor of this journal has been raised to 15.3). Importantly, Dr. Qin as the first author and Dr. Min as mentor won the Parmley Award in 2014 for the Young Author Achievement Award (Publication #2).

c) AIP1 in lymphangiogenesis: We have investigated the novel function of AIP1 in VEGFR-3 signaling, and VEGFR-3-dependent angiogenesis and lymphangiogenesis. AIP1, a signaling scaffold protein, is highly expressed in the vascular endothelium. We have previously reported that AIP1 functions as an endogenous inhibitor in pathological angiogenesis by blocking VEGFR-2 activity. Surprisingly, here we observe that mice with a global deletion of AIP1 (AIP1-KO) exhibit reduced retinal angiogenesis with less sprouting and fewer branches. Vascular endothelial cell (but not neuronal)-specific deletion of AIP1 causes similar defects in retinal angiogenesis. The reduced retinal angiogenesis correlates with reduced


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expression in VEGFR-3 despite increased VEGFR-2 levels in AIP1-KO retinas. Consistent with the reduced expression of VEGFR-3, AIP1-KO mice show delayed developmental lymphangiogenesis in neonatal skin and mesentery, and mount weaker VEGF-C-induced cornea lymphangiogenesis. In vitro, human lymphatic EC with AIP1 siRNA knockdown, retinal EC and lymphatic EC isolated from AIP1-KO all show attenuated VEGF-C-induced VEGFR-3 signaling. Mechanistically, we demonstrate that AIP1 increases VEGFR-3 protein expression, and by directly binding to VEGFR-3 enhances VEGFR-3 endocytosis and stability. Our in vivo and in vitro results provide the first insight into the mechanism by which AIP1 mediates VEGFR-3-dependent angiogenic and lymphangiogenic signaling (Publication #3). d). Lab members: There has been changes in lab members. 4 of them have taken professor positions in China. New personnel include Drs. Jenny Zhou, Mingzhu Yin, Lan Shao and Yeqi Wang (see photo). e). Collaborative research: We had very fruitful collaborative research in this year with VBT members or outside VBT. This is evident by 5 joint papers (Publications 4-8). f) On April 25, 2014, Dr. Min was invited to host the Nobel Laureate Forum with opening remarks in CMCB-2014, Zhe-Ao Bio Valley, Dalian, China (Speakers: Drs. Thomas C. Sudhof, Aaron Clechanover and Luc Montagnier). See attached photo with Dr. Sudhof, Nobel Laureate in Physiology/Medicine in 2013 3. Significance of Key Findings Relevant for the Mission of VBT: These studies provide insights into the mechanisms of vascular remodeling and tissue repair, providing potential new therapeutic targets for the treatment of vascular diseases. 4. Publications (Publications July 1, 2013– June 30, 2014): Al-Lamki RS, Lu W, Wang J, Yang J, Sargeant TJ, Wells R, Suo C, Wright P, Goddard M, Huang Q,

Lebastchi AH, Tellides G, Huang Y, Min W, Pober JS, Bradley JR (2013). TNF, Acting through Inducibly Expressed TNFR2, Drives Activation and Cell Cycle Entry of c-kit+ Cardiac Stem Cells in Ischemic Heart Disease. Stem Cells. (Epub ahead of print May 27, 2013).

Qin, L., Huang, Q., Zhang, H., Liu, R., Tellides, G., Min, W*, Yu, L*. (2014). SOCS1 prevents graft arteriosclerosis by preventing endothelial cell function. J Amer Coll of Cardiol. 63(1):21-9 (*Min is contacting senior author). Also see commentary in this issue by Weger, RA Immune regulators regulated to prevent transplant reactions. (Dr. Qin as the first author and Dr. Min as mentor won the Parmley Award in 2014 for the Young Author Achievement Award).

Zhou, HJ, Chen, X, Huang, Q., Zhang, H, Wang, Y, Yu, J, Liu, R, Li, Y, Xu, Z, and Min, W.* (2014) AIP1 mediates VEGFR3-dependent angiogenic and lymphangiogenic responses. Arterioscler Thromb Vasc Biol 34(3):603-15.

Kallen, AN., Xu, J., Qiao, C., Martinet, C., Lu, L., Ma, J., Zhou, XB., Yan, L., Liu, C., Yi, JS., Zhang, H., Min, W., Gregory, RI., Bennett, AM., Gabory, A., Dandolo, L., Huang, Y. (2013). H19 IncRNA acts as a natural sponge for let-7 miRNAs. Mol Cell. 52(1):101-12.

Zhang, Y., Tang, W., Zhang, H., Niu, X., Xu, Y., Zhang, J., Gao, K., Boggon, TJ., Toomre, D., Min, W.*, Wu, D.* (2013) A network of interactions enables CCM3 and STK24 coordinate unc13D-driven vesicle exocytosis in neutrophils. Dev Cell. 27(2):215-26. (*Min is co-corresponding author). PMCID: PMC3834565.

Wu, K., Liu, J., Tseng, SF, Gore, C., Sharifi, N., Fazli, L., Gleave, M., Kapur, P., Xiao, G., Sun, X, Oz, OK., Min, W., Alexandrakis, G., Yang, CR, Hsieh, CL, Wu, HC, He, D., Xie, D., Hsieh, JT. (2014) The role of DAB2IP in androgen receptor activation during prostate cancer progression. Oncogene. 33(15):1954-63.

Chen X, Zhou HJ, Huang Q, Lu L, Min W*. 2014) Novel action and Mechanism of Auranofin in Inhibition of Vascular Endothelial Growth Factor Receptor-3-Dependent Lymphangiogenesis. Anticancer Agents Med Chem. 14(7):946-954.


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Ji, W, Yang, M, Praggastis, A, Li Y, Zhou, HJ, He, Y, Ghazvinian, R, Cincotta, DJ, Rice, KP*, Min, W*.

(2014) Carbamoylating activity associated with the activation of the antitumor agent laromustine inhibits angiogenesis by inducing ASK1-dependent endothelial cell death. PLoS One 2014 Jul 28;9(7):e103224. doi:10.1371/journal.pone.0103224.


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Name: Laura E Niklason MD, PhD. Professor and Vice-Chair, Departments of Anesthesia & Biomedical Engineering

1. Overall Goal(s) of the Research Program of the Laboratory: Dr. Niklason’s research program focuses on vascular and lung tissue engineering, utilization of stem cells for tissue regeneration, and on potential mechanisms of aging in mammalian cells. 2. Specific Research Accomplishments in the last 12 months: Niklason’s group has continued to work on paradigms for allogeneic vascular engineering. Specifically, we have demonstrated the feasibility of using human iPS cells to produce mesenchymal stem cells and thence vascular smooth muscle cells. These iPS-derived cells, when cultured on biodegradable scaffolds inside bioreactors, produce vascular tissue containing SMC expressing contractile markers as well as collagen-based extracellular matrix. Ongoing work is seeking to enhance the differentiation state of iPS-derived SMC, as well as control for occasional “off-target” differentiation events to other mesenchymal lineages. In addition, we are developing novel covalent chemistries to modify the luminal surfaces of our vascular grafts, so as to reduce potential for thrombosis. Heparin-containing dendrimer molecules are covalently bound to graft collagens via amine and carboxyl groups, resulting in decreased platelet activation and inhibition of the coagulation cascade. Such chemical modifications may enhance the applicability of engineered grafts in small-diameter settings, where graft thrombosis is a primary failure mode. With respect to lung regeneration, we have advanced our approaches for cellular repopulation of acellular lung scaffolds. Working with human iPS cells, we have identified means of producing highly pure populations of distal lung epithelium expressing multiple markers of type II alveolar cells, including Nkx2.1 and SPC. In addition, we have developed novel assessments of lung matrix mechanics and permeability, to provide insights into microvascular functional patency and trans-alveolar capillary leak. Lastly, we have developed biomimetic bioreactors that can support the decellularization and subsequent culture of whole lung tissues from large animals and humans. This bioreactor provides vascular perfusion combined with negative- or positive-pressure breathing stimuli, that should assist with mass transfer and epithelial differentiation in cultured organs. In our aging project, we have assembled a list of putative genetic targets that may be involved in retardation of cellular aging phenomena. Confirmation of two of these targets for lifespan extension in C. elegans is underway. 3. Significance of Key Findings Relevant for the Mission of VBT: Related to the mission of VBT, our work in vascular regeneration is continuing to define the impact of physical forces on vessel growth, remodeling and mechanics. Our ongoing work on defining microvascular permeability in engineered lung models will enable us to develop experimental systmes that allow for direct testing of the role of vascular endothelium in lung barrier function.

4. Publications: (Publications July 1, 2013– June 30, 2014) Tsuchiya T, Balestrini JL, Mendez J, Calle EA, Zhao L, Niklason LE. Influence of pH on Extracellular

Matrix Preservation During Lung Decellularization. Tissue Eng Part C Methods. 2014 Jun 3. [Epub ahead of print] PubMed PMID: 24735501.

Calle EA, Ghaedi M, Sundaram S, Sivarapatna A, Tseng MK, Niklason LE. Strategies for whole lung tissue engineering. IEEE Trans Biomed Eng. 2014May;61(5):1482-96. doi: 10.1109/TBME.2014.2314261. Epub 2014 Mar 28. PubMed PMID:24691527; PubMed Central PMCID: PMC4126648.


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Jordan S. Pober, MD, PhD Bayer Professor of Translational Medicine; Professor and Vice-Chair, Department of Immunobiology, Section of Human and Translational Immunology

1. Overall Goal(s) of the Research Program of the Laboratory: The Pober laboratory studies the interactions of human immune and vascular systems. Our specific goals are to understand how vascular cells participate in and modify innate and adaptive immune responses; how the immune system can produce large vessel (ateriopathy) and small vessel (capillary leak) dysfunction, and how insights from these process may be applied to organ replacement therapy (transplant) and tissue engineering. 2. Specific Research Accomplishments in the last 12 months: Discoveries over the past 12 months include: (1) that CD4+ effector memory T cell transendothelial migration induced by endothelial antigen presentation requires T cell granule exocytosis and release of granzyme A; (2) that human alloantibody can activate the immunological functions of endothelial cells via a process involving deposition of complement membrane attack complex, clathrin-mediated internalization of membrane attack complex and activation of non-canonical NF-kB signaling on the surface of Rab5+ endosomes;(3) that capillary leak such as that involved in severe sepsis involves destabilization of inter-endothelial tight junctions in two discrete steps, both dependent upon activation of NF-kB; and (4) that pericytes provide both positive and negative signals that influence angiogenic sprouting of endothelial cells. . 3. Significance of Key Findings Relevant for the Mission of VBT: Our research can form the basis of new therapies for inflammation as well as for large and small vessel dysfunction. 4. Publications: (Primary Research Publications July 1, 2013– June 30, 2014) Andrejecsk JW, Cui J, Chang WG, Devalliere J, Pober JS, Saltzman WM. Exchanges of molecular signals

between alginate-encapsulated pericytes and freely suspended endothelial cells within a 3D protein gel. Biomaterials 2013; 34:8699-8908. PMCID: PMC3839675.

Jane-wit D, Manes TD, Yi T, Qin L, Clark P, Kirkiles-Smith NC, Abrahimi P, Devalliere J, Moeckel G, Kulkarni S, Tellides G, Pober JS. Alloantibody and complement promote T cell-Mediated cardiac allograft vasculopathy through non-canonical NF-κB signaling in endothelial Cells. Circulation 2013; 128:2504-2516. PMCID: PMC3885874.

Chang WG, Andrejecsk JW, Saltzman WM, Pober JS. Pericytes modulate endothelial sprouting. Cardiovasc Res. 2013; 100:492-500. PMCID: PMC3826704.

Devalliere J, Chang WG, Andrejecsk JW, Abrahimi P, Cheng CJ, Jane-wit D, Saltzman WM, Pober, JS. Sustained delivery of proangiogenic microRNA-132 by nanoparticle transfection improves endothelial cell transplantation. FASEB J. 2014: 28:908-922. PMCID: PMC3898640.

Lauridsen H, Pober JS, Gonzalez AL. A composite model of the human postcapillary venule for investigation of microvascular leukocyte recruitment. FASEB J. 2014; 28:1166-1180. PMCID: PMC3929680.

Spock CL, Tom LK, Canadas K, Sue GR, Sawh-Martinez R, Maier CL, Pober JS, Galan A, Schultz B, Waner M, Narayan D, Licinio J, Wong ML. Infantile hemangiomas exhibit neural crest and pericyte markers. Annals of Plastic Surgery 2014; (e-pub ahead of print).

Wang C, Qin L, Manes TD, Kirkiles-Smith NC, Tellides G, Pober JS. Rapamycin antagonizes TNF induction of VCAM-1 on endothelial cells by inhibiting mTORC2. J Exp Med. 2014; 211:395-404. PMCID: PMC3949571.


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Yibing Qyang, Ph.D. Assistant Professor of Medicine and Pathology

1. Overall Goal(s) of the Research Program of the Laboratory: Our research laboratory is interested in illuminating the underlying basic science and translational opportunities relevant to heart development and dysfunction. Our efforts center on a novel population of cardiovascular progenitor cells that are able to produce the majority of cell types comprising mouse heart and blood vessel tissues, using murine embryos and murine embryonic stem (ES) cells as experimental systems. Additionally, we are using these cardiovascular progenitor cells to generate 3D engineered cardiac tissues in order to repair the injured heart in mouse and rat models. Another focus of our lab is the use of induced pluripotent stem (iPS) cells to develop novel experimental models of human genetic diseases for the purpose of elucidating causative mechanisms and identifying potential therapeutic interventions to treat those diseases. Through a close collaboration with several clinicians at Yale, we are able to obtain cells from a variety of tissues procured from patients with cardiovascular diseases. These cells include dermal fibroblast cells derived from skin punch biopsies, which are isolated and reprogrammed into iPS cells in our laboratory before being re-differentiated into mature cardiovascular cells. In this way, we have the ability to derive an unlimited amount of cardiovascular cells containing disease-causing genetic errors for use in our investigations into the specifics of cardiovascular disease mechanisms. In addition to identification of the mechanisms responsible for disease phenotypes, we also have interest and experience in producing candidate molecular intervention strategies using small molecule screening and homologous recombination-mediated gene correction. Using a multidisciplinary approach to the study of cardiac development, cardiac physiology, stem cell biology and small molecule screening, we hope to contribute to the understanding of cardiovascular disease mechanisms as well as the development of novel therapeutic interventions for these diseases. 2. Specific Research Accomplishments in the last 12 months: We are reprogramming skin or blood cells from patients with cardiovascular diseases into pluripotent stem cells and then redifferentiate these stem cells into mature cardiovascular cells. In this way, we will be able to derive unlimited amount of cardiovascular cells with disease-causing mutations and study cardiovascular disease mechanisms. Specifically, we discovered a FDA-approved chemotherapeutic agent, vinblastine, which induced formation of actin filament bundles and inhibited hyperproliferation of smooth muscle cells (SMCs) derived from SVAS iPSCs. We will obtain mechanistic insight into how elastin and vinblastine inhibit the hyperproliferation of SVAS and WBS iPSC-SMCs and to screen for additional small molecules that ameliorate the hyperproliferative defect in SVAS and WBS and other vascular proliferative diseases. Heart failure caused by myocardial infarction remains a leading cause of morbidity and mortality in the developed world. We have established scaffold-free engineered heart tissues (EHT) with CPC and examine their contribution to heart repair and regeneration in animal models. Our studies have provided the first evidence that ESC-derived ISL1+ CPCs can effectively form new cardiac muscle in vitro and in vivo and improve heart function after implantation. ISL1+ CPCs will provide an abundant renewable cell source for basic research and will set the stage for using them for cell-based therapies for heart failure. Vascular disease due to arterial stenosis is the largest cause of mortality in the developed world. We have recently derived unlimited amounts of highly homogeneous functional vascular SMCs from hiPSCs (hiPSC-SMCs) and will investigate the therapeutic potential of hiPSC- and hESC-SMCs by developing TEBVs using biodegradable polyglycolic acid (PGA) scaffolds in a pulsatile bioreactor system and implanting them as aortic interposition grafts in nude rats. Development of TEBVs with significantly improved compliance using hiPSC-SMCs will enable us to take the next step towards developing autologous tissue engineered grafts for clinical intervention in vascular diseases.


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3. Significance of Key Findings Relevant for the Mission of VBT

Generation of cardiovascular disease models using induced pluripotent stem (iPS) cells derived from patient skin fibroblast cells and screening of small molecules that can rescue disease phenotypes. Diseases currently studied: Supravalvular aortic stenosis (SVAS), Williams syndrome (WBS), Down’s syndrome and hypertrophic cardiomyopathy.

Derivation of cardiovascular cells from human embryonic stem (ES) and iPS cells, establishment of engineered heart tissues and blood vessels using these cells, and examination of their contribution to cardiovascular repair.

4. Publications: (Publications July 1, 2013 – June 30, 2014)

Li W, Li Q, Qin L, Ali R, Qyang Y, Tassabehji M, Pober BR, Sessa WC, Giordano FJ, Tellides G (2013). Rapamycin Inhibits Smooth Muscle Cell Proliferation and Obstructive Arteriopathy Attributable to Elastin Deficiency. Arterioscler Thromb Vasc Biol. 33(5):1028-35. PMID: 23493289

Dunworth WP, Cardona-Costa J, Cagavi E, Kim JD, Fischer JC, Meadows S, Wang Y, Cleaver O, Qyang Y, Ober EA, Jin SW (2013). Bone Morphogenetic Protein 2 Signaling Negatively Modulates Lymphatic Development in Vertebrate Embryos. Circ Res. 2014 Jan 3;114(1):56-66. PMID: 24122719

Suh CY, Wang Z, Bártulos O, Qyang Y (2014). Advancements in Induced Pluripotent Stem Cell Technology for Cardiac Regenerative Medicine. J Cardiovasc Pharmacol Ther. 2014 Mar 19;19(4):330-339. PMID: 24651517


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Nancy Hartman Ruddle, Ph.D. Professor Emeritus of and Senior Research Scientist in Epidemiology (Microbial Diseases) 1. Overall Goal(s) of the Research Program of the Laboratory: We study cell trafficking and inflammation in autoimmunity and lymphoid organ development, particularly the roles of members of the lymphotoxin/tumor necrosis factor (LT/??TNF) family. We study acute inflammation and animal models of autoimmune diseases, including Type 1 diabetes mellitus and multiple sclerosis. LT is also crucial for lymphoid organ development; LT deficient mice lack lymph nodes and Peyer’s patches and exhibit profound alterations in spleen and nasal associated lymphoid tissue. Our studies demonstrate that cytokines’ functions in lymphoid organ development and inflammation are similar; they regulate chemokines and vascular adhesion molecules. Ectopic or “tertiary” lymphoid organs arising in chronic inflammation are lymphoid accumulations that permit the presentation of foreign and self-antigens at local sites of inflammation. Our studies on high endothelial venules and lymphatic vessels elucidate developmental mechanisms and point the way towards treatment and prevention of chronic inflammation. 2. Publications: (Publications July 1, 2013– June 30, 2014) Ruddle NH. Lymphotoxin and TNF: how it all began-a tribute to the travelers. Cytokine Growth Factor

Rev. 2014 Apr;25(2):83-9. doi:10.1016/j.cytogfr.2014.02.001. Epub 2014 Feb 12. PubMed PMID: 24636534; PubMedCentral PMCID: PMC4027955.

Viehmann Milam AA, Maher SE, Gibson JA, Lebastchi J, Wen L, Ruddle NH, Herold KC, Bothwell AL. A humanized mouse model of autoimmune insulitis. Diabetes. 2014 May;63(5):1712-24. doi: 10.2337/db13-1141. Epub 2014 Jan 29. PubMed PMID:24478396; PubMed Central PMCID: PMC3994947.

Truman LA, A-Gonzalez N, Bentley KL, Ruddle NH. Lymphatic vessel function inhead and neck inflammation. Lymphat Res Biol. 2013 Sep;11(3):187-92. doi:10.1089/lrb.2013.0013. PubMed PMID: 24044758; PubMed Central PMCID: PMC3780307.


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Mehran M. Sadeghi, M.D. Associate Professor of Medicine (Cardiology)

1. Overall Goal(s) of the Research Program of the Laboratory: The main goal of our research is to develop novel molecular imaging approaches for cardiovascular diseases, with an emphasis on the vascular system. We have focused on four examples of vascular remodeling, namely injury-induced vascular remodeling, graft arteriosclerosis, aneurysm, and atherosclerosis. For each process studied, we identify specific imaging targets based on pathophysiology or genomic and proteomic screening, develop novel ligands for imaging or use existing radiotracers, establish relevant animal models, and use small animal imaging systems [Single-photon emission computed tomography (SPECT), CT, Near Infrared Fluorescence(NIRF)] to detect and quantify the process in vivo. Studies of the pathophysiology of vascular remodeling, to identify novel targets for imaging and to understand the biology of the targets identified, is an integral part of our research. 2. Specific Research Accomplishments in the last 12 months: We have demonstrated that MMP-targeted imaging can detect and monitor the effect of therapeutic interventions, including dietary modification and lipid lowering therapies, in murine atherosclerosis. In a comparison of three interventions to lower blood lipid levels, we found a remarkable reduction in vessel wall inflammation following dietary intervention. Both fenofibrate and simvastatin reduced total blood cholesterol level, but fenofibrate appeared to have a more robust effect compared to simvastatin on vessel wall inflammation imaged and quantified in vivo. In addition using serial imaging in vivo we show major differences in subjects within the same group in regards to response to therapy. ESDN is a neuropilin-like protein that we identified as a potential target for imaging vascular remodeling. Investigating the role of ESDN in vascular biology, we showed that ESDN regulates VEGF signaling in endothelial cells. In addition, VEGF responses as well as developmental and ischemic angiogenesis are impaired in the ESDN knockout mouse, generated in our laboratory. Similarly, ESDN downregulation affects vascular development in zebrafish. We linked the observed effect of ESDN to its regulation of the VEGFR-2/ protein tyrosine phosphatase association. As such, ESDN may be a novel therapeutic target for regulating angiogenesis. 3. Publications: (Publications July 1, 2013 – June 30, 2014) Nie L, Guo X, Esmailzadeh L, Zhang J, Asadi A, Collinge M, Kim DJ, Jin SW, Dubrac A, Eichmann A,

Simons M, Bender JR, Sadeghi MM. “Transmembrane protein ESDN promotes endothelial VEGF signaling and regulates angiogenesis”. Journal of Clinical Investigation, 2013, 123(12):5082-97. PMC: 3859420

Golestani R, Sadeghi MM. “Emergence of molecular imaging of aortic aneurysm; implications for risk stratification and management”. Journal of Nuclear Cardiology, 2014, 21(2):251-67.

Razavian M, Nie L, Challa A, Zhang J, Golestani R, Jung JJ, Robinson S, Sadeghi MM. “Lipid lowering and imaging protease activation in atherosclerosis”. Journal of Nuclear Cardiology, 2014, 21(2):319-28.


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W. Mark Saltzman Goizueta Foundation Professor of Chemical and Biomedical Engineering

1. Overall Goal(s) of the Research Program of the Laboratory: Our laboratory is creating new technology, based on the use of biocompatible polymeric materials, for the controlled delivery of drugs, proteins, and genes. We also develop and study new polymeric materials that influence the growth and assembly of tissues. Our research projects in the area of tissue engineering are the most relevant to the VBT program. Tissue engineering is a new field of inquiry, defined about 15 years ago. In our view tissue engineering involves the use of synthetic polymers as scaffolds for cell transplantation, in which the properties of the scaffold are tuned to encourage the formation or regeneration of tissue structure and function. Importantly, tissue engineering involves a combination of disciplines to achieve new therapies and, in some cases, entirely new approaches to therapy. 2. Specific Research Accomplishments in the last 12 months: We are particularly interested in developing methods for transplantation of neo-tissues—combinations of cells and synthetic materials that are assembled ex vivo. In the past year, we have made progress in several areas that are important in this overall effort. First, we have developed techniques for the synthesis of nanoparticles that are loaded with plasmids and oligonucleotides for gene therapy, knockdown, and editing. Second, we have developed versatile methods for cell encapsulation, which allows us to use cells as permanent sources of protein release and delivery. Third, we have developed electrospun fiber meshes that can be used as scaffolds for cell growth, and further modified to provide for gene delivery. 3. Significance of Key Findings Relevant for the Mission of VBT: We are committed to collaboration with VBT scientists on projects relevant for the VBT mission. Recently, we worked with Themis Kyriakides to use these materials for the sustained delivery of plasmid DNA to aid in wound healing. Second, we have developed encapsulation systems for VEGF, other protein growth factors, and cells: we are using these in collaboration with Jordan Pober, to optimize endothelial cell transplantation for treatment of ischemia and tissue engineering. Third, we have developed nanoparticles that can produce sustained delivery of siRNA or microRNA in cells: we are using these collaboration with Jordan Pober and George Tellides to provide targeted delivery to endothelial cells in transplanted tissues, as a new means of preventing transplant rejection. 4. Publications: (Publications July 1, 2013– June 30, 2014) Kobsa S, Kristofik NJ, Sawyer AJ, Bothwell A, Kyriakides TR, and Saltzman WM. An electrospun scaffold

integrating nucleic acid delivery for treatment of full thickness wounds, Biomaterials 34:3891-3901 (2013). PMCID: PMC3625647

Andrejecsk JW, Cui J, Chang WG, Devalliere J, Pober JS, Saltzman WM. Paracrine exchanges of molecular signals between alginate-encapsulated pericytes and freely suspended endothelial cells within a 3D protein gel, Biomaterials 35(5):8899-8908 (2013). PCMID: PMC3839675

Cheng CJ, Saltzman WM, Slack FJ. Canonical and non-canonical barriers facing antimiR cancer therapeutics. Curr Med Chem 20:3582-3593 (2013). PMCID: PMC3901840.

Chang WG, Andrejecsk JW, Kluger MS, Saltzman WM, and Pober JS. Pericytes modulate endothelial sprouting. Cardiovascular Research 100:492-500 (2013). PMCID: PMC3826704.

Schleifman EB, McNeer NA, Jackson A, Yamtich J, Leif J, Shultz LD, Greiner DL, Kumar P, Saltzman WM, and Glazer PM. Site-specific genome editing in PBMCs with PLGA nanoparticle-delivered PNAs confers HIV-1 resistance in humanized mice, Molecular Therapy-Nucleic Acids, Nov 19;2:e135. doi: 10.1038/mtna.2013.59 (2013). PMCID: PMC3889188

Saltzman WM and Kyriakides TR. Cell interactions with polymers In Principles of Tissue Engineering, Fourth

edition, RP Lanza, R Langer, J Vacanti (Ed.), Academic Press, NY, p. 385-406 (2014).


46

Look M, Saltzman WM, Craft J, and Fahmy TM. Nanomaterial-dependent modulation of dendritic cells

affects the extent of therapeutic immunosuppresion in lupus, Biomaterials 35:1089-1095 (2014). PMCID: in progress.

Devalliere J, Chang WG, Andrejecsk JW, Cheng CJ, Jane-wit D, Saltzman WM, and Pober JS. Sustained delivery of pro-angiogenic microRNA-132 by nanoparticle transfection improves endothelial cell transplantation, FASEB Journal 28(2): 908-922 (2014). PMCID: PMC3898640.

Martin DT, Steinbach JM, Liu J, Shimizu S, Kaimakliotis HZ, Wheeler MA, Hittelman AB, Saltzman WM, Weiss RM. Surface modified nanoparticles enhance transurothelial penetration and delivery of survivin siRNA in treating bladder cancer, Molecular Cancer Therapeutics 13:71-81 (2014). PMCID: in process

Gattu A, Birkenfeld A, Iwakiri Y, Jay SM, Saltzman WM, Doll J, Protiva P, Jornyvaz F, Samuel V, Crawford SE, and Chung C. Pigment Epithelium-Derived Factor (PEDF) suppresses IL-1beta-mediated c-Jun N-terminal Kinase (JNK) activation to improve hepatocyte insulin signaling, Endocrinology 155:1373-85 (2014). PMCID: in process.

Deng Y, Saucier-Sawyer JK, Hoimes CJ, Zhang J, Seo YE, Andrejecsk JW, and Saltzman WM. The effect of hyperbranched polyglycerol coatings on drug delivery using degradable polymer nanoparticles. Biomaterials 35:6595-6602 (2014). PMCID: in process.

Sawyer AJ, Tian W, Saucier-Sawyer JK, Rizk PJ, Saltzman WM, Bellamkonda RV, Kyriakides TR. The effect of inflammatory cell-derived MCP-1 loss on neuronal survival during chronic inflammation, Biomaterials 35:6698-6706 (2014). PMCID: in process.

Ricciardi AS, McNeer NA, Anandalingam K, Saltzman WM, and Glazer PM. Targeted genome modification via triple helix formation, Methods in Molecular Biology 1176:89-106 (2014).

Weiser JR and Saltzman WM. Controlled release for local delivery of drugs: Barriers, models, and opportunities, Journal of Controlled Release, in press.

Gupta R, Dong Y, Solomon PD, Wettersten HI, Cheng CJ, Min JN, Henson J, Dogra SK, Hwang SH, Hammock BD, Zhu LJ, Reddel RR, Saltzman WM, Weiss RH, Chang S, Green MR, Wajapeyee N, Synergistic tumor suppression by combined inhibition of telomerase and CDKN1A, Proceedings of the National Academy of Sciences, in press.

Bahal R, Quijano E, McNeer NA, Liu Y, Bhunia DC, López F, Saltzman WM, Ly DH, Glazer PM. Single-stranded γPNAs for in vivo site-specific genome editing via Watson-Crick recognition, Current Gene Therapy, in press.

Ediriwickrema A, Zhou J, Deng Y, and Saltzman WM. Multi-layer nanoparticles for combinatorial gene and drug delivery to tumors, Biomaterials, in press.


47

Martin Schwartz, Ph.D. Robert W. Berliner Professor of Medicine (Cardiology and Professor of Cell Biology

1. Overall Goal(s) of the Research Program of the Laboratory: Our laboratory studies signaling by integrins and mechanotransduction in the vascular system. We are especially interested in how endothelial cells respond to forces from flowing blood. The goals are to understand basic mechanisms of signal transduction, and to apply this information to both atherosclerosis and flow-dependent artery remodeling. 2. Specific Research Accomplishments in the last 12 months: We have elucidated an interaction between the VE-cadherin VEGF receptors through their transmembrane domains that is essential for flow sensing. We have determined that vascular remodeling is governed by a fluid shear stress set point, found that this set point varies between different types of vessels where flow patterns are different, and a mechanism that determines these variations in the set point. Lastly, we have made major progress toward understanding the link between extracellular matrix remodeling and inflammation as occurs in vascular remodeling and atherosclerosis. 3. Significance of Key Findings Relevant for the Mission of VBT: These results advance our understanding of basic mechanisms by which endothelial cells respond to fluid shear stress and the downstream pathways by which flow triggers vascular remodeling and atherosclerosis. 4. Publications: (Publications July 1, 2013– June 30, 2014) Ross TD, Coon BG, Yun S, Baeyens N, Tanaka K, Ouyang M, Schwartz MA. (2013) Integrins in

mechanotransduction. Curr Opin Cell Biol. 25(5):613-8. Wang C, Baker BM, Chen CS, Schwartz MA. (2013) Endothelial cell sensing of flow direction. Arterioscler

Thromb Vasc Biol. 33(9):2130-6. Dufresne ER, Schwartz MA. (2013) Cell migration: Towards the void. Nat Mater. 12(9):783-4. Conway DE, Schwartz MA. (2013) Flow-dependent cellular mechanotransduction in atherosclerosis. J

Cell Sci. 126:5101-9. Moissoglu K, Kiessling V, Wan C, Hoffman B, Norambuena A, Tamm L, and Schwartz MA. (2014)

Regulation of Rac translocation and activation by membrane domains and their boundaries. J Cell Sci. 127:2565-76.

Humphrey, JD, Milewicz, DM, Tellides G, Schwartz MA (2014) Cell Biology Dysfunctional mechanosensing in aneurysms. Science 2 May 2014, 344:477-9.

Schwartz, MA. (2014) Sticking to it: Tracking the paths of integrin signaling. Nat Cell Biol. May 30;16(6):487.

Schwartz, MA. (2014) Fluid shear stress on endothelial cells modulates mechanical tension across VE-cadherin and PECAM-1. Cell Adh Migr., Volume 9, Issue 2. (invited review) in press.

Leerberg JM, Gomez GA, Verma S, Moussa EJ, Wu SK, Priya R, Hoffman BD, Grashoff C, Schwartz MA, Yap AS. (2014) Tension-sensitive actin assembly supports contractility at the epithelial zonula adherens. Curr Biol. In press.

Moissoglu K, Schwartz MA. (2014) Spatial and temporal control of Rho GTPase functions. Cell Logistics, In press.


48

William C. Sessa, Ph.D. Director Vascular Biology & Therapeutics Program, Vice Chairman Pharmacology, Alfred Gilman Professor of Pharmacology and Professor of Medicine (Cardiology) 1. Overall Goal (s) of the Research Program of the Laboratory: Our laboratory is very interested in endothelial cell biology, signaling and regulation of post-natal angiogenesis/ arteriogenesis and atherosclerosis. 2. Specific accomplishments in the last year: In the past year, we have made successful inroads into several areas of nitric oxide signaling, caveolin function and miRNAs. 3. Publications: (Publications July 1, 2013– June 30, 2014) Lee MY, Luciano AK, Ackah E, Rodriguez-Vita J, Bancroft TA, Eichmann A, Simons M, Kyriakides TR,

Morales-Ruiz M, Sessa WC. Endothelial Akt1 mediates angiogenesis by phosphorylating multiple angiogenic substrates. Proc Natl Acad Sci U S A. 2014 Sep 2;111(35):12865-70. doi: 10.1073/pnas.1408472111. Epub 2014 Aug 18. PubMed PMID: 25136137.

Park EJ, Grabińska KA, Guan Z, Stránecký V, Hartmannová H, Hodaňová K, Barešová V, Sovová J, Jozsef L, Ondrušková N, Hansíková H, Honzík T, Zeman J, Hůlková H, Wen R, Kmoch S, Sessa WC. Mutation of Nogo-B Receptor, a Subunit of cis-Prenyltransferase, Causes a Congenital Disorder of Glycosylation. Cell Metab.2014 Sep 2;20(3):448-57. doi: 10.1016/j.cmet.2014.06.016. Epub 2014 Jul 24. PubMed PMID: 25066056; PubMed Central PMCID: PMC4161961.

Mehra VC, Jackson E, Zhang XM, Jiang XC, Dobrucki LW, Yu J, Bernatchez P, Sinusas AJ, Shulman GI, Sessa WC, Yarovinsky TO, Bender JR. Ceramide-activated phosphatase mediates fatty acid-induced endothelial VEGF resistance and impaired angiogenesis. Am J Pathol. 2014 May;184(5):1562-76. doi:10.1016/j.ajpath.2014.01.009. Epub 2014 Mar 5. PubMed PMID: 24606881; PubMed Central PMCID: PMC4005977.

Lee MY, Skoura A, Park EJ, Landskroner-Eiger S, Jozsef L, Luciano AK, Murata T, Pasula S, Dong Y, Bouaouina M, Calderwood DA, Ferguson SM, De Camilli P, Sessa WC. Dynamin 2 regulation of integrin endocytosis, but not VEGF signaling, is crucial for developmental angiogenesis. Development. 2014 Apr;141(7):1465-72. doi: 10.1242/dev.104539. Epub 2014 Mar 5. PubMed PMID: 24598168; PubMed Central PMCID: PMC3957370.

Kondo Y, Jadlowiec CC, Muto A, Yi T, Protack C, Collins MJ, Tellides G, Sessa WC, Dardik A. The Nogo-B-PirB axis controls macrophage-mediated vascular remodeling. PLoS One. 2013 Nov 20;8(11):e81019. doi:10.1371/journal.pone.0081019. eCollection 2013. PubMed PMID: 24278366; PubMed Central PMCID: PMC3835671.

Ju R, Zhuang ZW, Zhang J, Lanahan AA, Kyriakides T, Sessa WC, Simons M. Angiopoietin-2 secretion by endothelial cell exosomes: regulation by the phosphatidylinositol 3-kinase (PI3K)/Akt/endothelial nitric oxide synthase (eNOS) and syndecan-4/syntenin pathways. J Biol Chem. 2014 Jan 3;289(1):510-9. doi:10.1074/jbc.M113.506899. Epub 2013 Nov 14. PubMed PMID: 24235146; PubMed Central PMCID: PMC3879572.

Di Lorenzo A, Lin MI, Murata T, Landskroner-Eiger S, Schleicher M, Kothiya M, Iwakiri Y, Yu J, Huang PL, Sessa WC. eNOS-derived nitric oxide regulates endothelial barrier function through VE-cadherin and Rho GTPases. J Cell Sci. 2013 Dec 15;126(Pt 24):5541-52. doi: 10.1242/jcs.115972. Epub 2013 Sep 17. Erratum in: J Cell Sci. 2014 May 1;127(Pt 9):2120. PubMed PMID: 24046447; PubMed Central PMCID: PMC3860306.

Huang Y, Di Lorenzo A, Jiang W, Cantalupo A, Sessa WC, Giordano FJ. Hypoxia-inducible factor-1α in vascular smooth muscle regulates blood pressure homeostasis through a peroxisome proliferator-activated receptor-γ-angiotensin II receptor type 1 axis. Hypertension. 2013 Sep;62(3):634-40. doi:10.1161/HYPERTENSIONAHA.111.00160. Epub 2013 Aug 5. PubMed PMID: 23918749.


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Zhang P, Huang B, Xu X, Sessa WC. Ten-eleven translocation (Tet) and thymine DNA glycosylase (TDG),

components of the demethylation pathway, are direct targets of miRNA-29a. Biochem Biophys Res Commun. 2013 Aug 2;437(3):368-73. doi: 10.1016/j.bbrc.2013.06.082. Epub 2013 Jun 29. PubMed PMID: 23820384; PubMed Central PMCID: PMC3767426.


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Michael Simons, M.D. RW Berliner Professor of Medicine & Cell Biology Director, Yale Cardiovascular Research Center

1. Overall Goals of the Research Program of the Laboratory Our laboratory is interested in regulation of arterial and lymphatic morphogenesis and angiogenic growth factor signaling. These process are investigated at all levels, including in vitro signaling studies, in vivo mouse transgenic and knock-out models and translational studies in larger animal models and early phase clinical trials. 2. Specific Research Accomplishments in the last 12 months We have made significant advances in unraveling the role of VEGF and FGF signaling in the normal endothelium, novel mechanism of endothelial-to-mesenchyma transition and key molecular pathways regulating arteriogenesis and lymphangiogenesis. 3. Significance of Key Findings Relevant for the Mission of VBT These findings advance our knowledge of molecular details of regulation of vascular development and signaling and should eventually enable the development of new therapeutic paradigms. 4. Key Publications: (Publications July 1, 2013– June 30, 2014) Moraes F, Paye J, Mc Gabhann F, Zhuang ZW, Zhang J, Lanahan AA, Simons M. Endothelial cell-

dependent regulation of arteriogenesis. Circ Res 2013; 113:1076-86; doi: 10.1161/113.301340 PMID 23897694 (10 most read Circ Res papers of 2013)

Ding B-S, Cao Z, Lis R, Nolan DJ, Guo P, Simons M, Penfold M, Shido K, Rabbany SY, Rafii S. Divergent signals from vascular niche balance liver regeneration and fibrosis. Nature 2014; 505:97-102; doi: 10.1038/nature12681. PMID: 24256728

Ju R, Zhuang ZW, Zhang J, Lanahan AA, Kyriakides T, Sessa WC, Simons M. Angiopoietin-2 secretion by endothelial cells exosome: regulation by the PI3K/Akt/eNOS and syndecan 4/syntenin pathways. J Biol Chem 2014; 289:510-519 doi:10.1074/jbc.M113.506899 PMID: 24235146

Sakurai T, Woolls MJ, Jin SW, Murakami M, Simons M. Inter-Cellular Exchange of Cellular Components via VE-Cadherin-Dependent Trans-Endocytosis. PLoS One. 2014 9(3):e90736. doi: 10.1371/journal.pone.0090736. eCollection 2014.

Cao Z, Ding BS, Guo P, Lee SB, Butler JM, Casey SC, Simons M, Tam W, Felsher DW, Shido K, Rafii A, Scandura JM, Rafii S. Angiocrine factors deployed by tumor vascular niche induce B cell lymphoma invasiveness and chemoresistance. Cancer Cell. 2014; 25(3):350-65. doi: 10.1016/j.ccr.2014.02.005. PMID: 24651014

Chen P-Y, Qin L, Zhuang ZW, Tellides G, Lax I, Schlessinger J, Simons M. The docking protein FRS2α is a critical regulator of VEGF receptors signaling. Proc Natl Acad Sci USA 2014; 111:5514-19. doi: 10.1073/pnas.1404545111; PMID:24706887

Brunet I, Gordon E, Han J, Cristofaro B, Broqueres-You D, Liu C, Zhang J, del Toro R, Mathivet T, Larrivée B, Jagu J, Bouvrée K, Pardanaud L, Machado MJC, Kennedy TE, Simons M, Zhuang ZW, Levy BI, Tessier-Lavigne M, Grenz A, Eltzschig H and Eichmann A. Netrin-1 controls sympathetic innervation. J Clin Invest 2014; 124(7):3230-40. doi: 10.1172/JCI75181 doi: 10.1172/JCI75181. PMID: 24937433

Morrison AR, Yarovinsky TO, Young BD, Moraes F, Ross TD, Ceneri N, Zhang J, Lanahan AA, Lech D, Dubrac A, Zhang J, Zhang ZW, Eichmann E, Simons M. PTP1b is a physiologic regulator of VEGF signaling in endothelial cells. Circulation 2014; doi: 10.1161/CIRCULATIONAHA.114.009683.


51

Albert Sinusas, MD Professor of Medicine (Cardiology) and Diagnostic Radiology; Director, Cardiovascular Imaging; Director, Yale Translational Research Imaging Center (Y-TRIC)

1. Overall Goal(s) of the Research Program of the Laboratory: Research in the Sinusas laboratory is directed at development of noninvasive imaging approaches for the assessment of myocardial viability, angiogenesis, arteriogenesis, and post-infarction remodeling. The laboratory has been employing the 3-D modalities of single photon emission computed tomography (SPECT), positron emission tomography (PET), echocardiography, X-ray tomography, and magnetic resonance (MR) imaging for assessment of a wide range of physiological and molecular processes primarily focused in the cardiovascular system. The laboratory is currently focused on targeted molecular imaging, developing non-invasive nuclear imaging strategies for identifying the hypoxic stimulus for angiogenesis, and targeted imaging of selected integrins previously established to modulate the angiogenic process, and the interrelationship of angiogenesis and arteriogenesis. These studies involve the use of rodent and large animal models of myocardial ischemia as well as hindlimb ischemia. The lab is also focused on development of novel biodegradable intravascular stents and imaging devices and the evaluation of tissue engineered grafts. 2. Specific Research Accomplishments in the last 12 months: Application of quantitative tools for automated physiologically based segmentation of the coronary and peripheral vasculature. Development of CT and MR approached for evaluation of angiogenesis, tissue oxygenation and arteriogenesis. Development of novel multimodality nanoparticle-based contrast agents. Developed novel prototype biodegradable magnetizable coronary stent for attraction of stem cells to prevent restenosis and late in stent thrombosis. 3. Significance of Key Findings Relevant for the Mission of VBT: Demonstrated the phenomenon of augmented CT attenuation due to nano-confinement with the development of novel CT containing multimodality imaging agents. This development has significant implications for targeted molecular CT-based imaging of the vascular tree. Development of prototype intravascular stents. 4. Publications: (Publications July 1, 2013– June 30, 2014) Stacy MR, Yu DY, Maxfield MW, Jaba IM, Jozwik BP, Zhuang ZW, Lin BA, Hawley CL, Caracciolo CM, Pal

P, Tirziu D, Sampath S, Sinusas AJ. Multimodality Imaging Approach for Serial Assessment of Regional Changes in Lower Extremity Arteriogenesis and Tissue Perfusion in a Porcine Model of Peripheral Arterial Disease. Circ Cardiovasc Imaging. 2013 Oct 29. [Epub ahead of print] PMID: 24170237

Stacy MR, Naito Y, Maxfield MW, Kurobe H, Tara S, Chan C, Rocco KA, Shinoka T, Sinusas AJ, Breuer CK. Targeted imaging of matrix metalloproteinase activity in the evaluation of remodeling tissue-engineered vascular grafts implanted in a growing lamb model. J Thorac Cardiovasc Surg. 2014 May 21:S0022-5223(14)00581-9. PMID:24952823

Kurobe H, Maxfield MW, Naito Y, Cleary M, Stacy MR, Solomon D, Rocco KA, Tara S, Lee AY, Sinusas AJ, Snyder EL, Shinoka T, Breuer CK. Comparison of a Closed System to a Standard Open Technique for Preparing Tissue-Engineered Vascular Gifts. Tissue Eng. Part C Methods. 2014 Jul 3. PMID:24866863

Mehra VC, Jackson E, Zhang XM, Jiang XC, Dobrucki LW, Yu J, Bernatchez P, Sinusas AJ, Shulman GI, Sessa WC, Yarovinsky TO, Bender JR. Ceramide-Activated Phosphatase Mediates Fatty Acid-Induced Endothelial VEGF Resistance and Impaired Angiogenesis. Am J Pathol. 2014; 184(5):1562-76 PMID: 24606881


52

Edward L. Snyder, M.D. Professor Laboratory Medicine, Director, Apheresis/Cell Processing VBT Core Facility (Core D)

1. Overall Goal(s) of the Research Program of the Laboratory: The Apheresis/Cell Processing Core Facility played a critical role in the Vascular Biology and Transplantation Program. The Cell Processing Core Laboratory is designed to support the needs of the VBT Program users who are performing basic science and clinical research involving mononuclear and other cell types, by providing five specific functions. First, the pheresis section of the Cell Processing Core Laboratory procured and provided normal donor specimens in support of research projects. These samples, obtained under IRB approved protocols from fresh specimens, were made available to VBT membership. Second the VBT Core D Cell Collection and Processing Laboratory can provide, as requested, cell purification services. Third, the Cell Processing Core can provide large-scale processing capabilities in support of specific research studies involving human MNCs as well as CD34 positive and other cell types. Included within this section was the development of cell selection and culturing techniques to support the novel cell therapy protocols, as well as the pre-clinical validation of research procedures. The VBT Core resource provided the critical instrumentation and technical expertise in cell processing and cryopreservation, needed for the in vitro use of cells, or infusion of cells into animals. Fourth, the Core provided collections of MNC and could as, and if, needed, provide CD34+ cells from G-CSF stimulated donors. Fifth, the Apheresis/ Cell Processing VBT Core Facility maintained compliance with institutional, NIH, FDA, and AABB guidelines, and ensured that the protocols were safely and effectively applied. This objective includes training of new investigators in Compliance and Quality Control issues. Thus, this resource provided access to cell collection, selection, processing and culturing technologies, as well as services and scientific consultation to enhance the productivity of the VBT members. This technically sophisticated resource was critical to the VBT Section’s research progress. 2. Specific Accomplishments in the last 12 months: In 2013 - 2014, Core D performed 19 MNC apheresis collections for Program Leaders’ research. 3. Key Findings: N/A -- Core Facility 4. Publications: (Publications July 1, 2013– June 30, 2014) Devine S. Optimizing autologous stem cell mobilization strategies to improve patient outcomes: consensus

guidelines and recommendations. BBMT 2014 Mar; 20(3):295-308. Kurobe, H, Maxfield MW, Naito Y, Cleary M, Stacy MR, Solomon D, Rocco KA, Tara S, Lee AY, Sinusas

AJ, Snyder, EL., Shinoka T, Breuer, CK. Comparison of a closed system to a standard open technique for preparing tissue engineered vascular grafts. Tissue Engineering Part C, Methods, 2014 (epublished ahead of print).

Burns LJ, Gajewski JL, Majhail NS, Navarro W, Perales MA, Shereck E, Selby GB, Snyder, EL, Woolfrey AE, Litzow, MR. Challenges and potential solutions for recruitment and retention of hematopoietic cell transplant physicians: the NMDP System Capacity Initiative: Physician Workforce Group report. BBMT 2014 May,20(5):617-21.

Alhumaidan H, Perrotta PL, Han Y, Snyder EL. Blood Transfusion. In: Oxford Textbook of Medicine, Firth J, Cox T, Warrell D, eds. 5th edition – Online Updates, Oxford University Press, 2013.


53

Bing Su, Ph.D. Associate Professor of Immunobiology 1. Overall Goal(s) of the Research Program of the Laboratory: The overall goals of the research program of this laboratory are to understand the biology of signal transduction mediated by the mitogen-activated protein kinase (MAPK) pathway, and the mechanistic target of rapamycin (mTOR) pathway. We focus on the cardiovascular function and immune function of these two signal transduction pathways, and we utilize approaches of genetics, biochemistry, and molecular biology. We would like to understand the molecular mechanisms of MEKK2/3 mediated MAPK activation and the Sin1 mediated mTOR regulation in the immune cell development and differentiation, and in endothelial cell function in angiogenesis and blood vessel integrity. 2. Specific Research Accomplishments in the last 12 months: In the past 12 months, we investigate the roles of MEKK2 in inflammatory bowel diseases using MEKK2 KO mice. We found that MEKK2 could protect mice from DSS induced intestinal inflammation. At the molecular levels, we found that the TLR2/TLR6 mediated inflammatory signals require MEKK2. We also found evidence that MEKK2 may participate in inflammasome activation, which we are verifying these data. We continue the study of MEKK3 in brain endothelial cells in regulating the brain angiogenesis and BBB integrity. We demonstrate that MEKK3 plays an essential role in these processes. In collaboration with Titus Boggon, we revealed the critical amino acids in both MEKK3 and CCM2 that are required for their interaction. We are testing the idea that this interaction is indeed critical for the endothelial function in the brain for BBB integrity. In addition, we investigate the role of Sin1-mTORC2 in angiogenesis and development. We have generated Sin1floxed mice successfully and we are in the process of analyzing the tissue specific Sin1 KO mice in endothelial cells and in lymphocytes. 3. Significance of Key Findings Relevant for the Mission of VBT: The MAPK and mTOR pathways are crucial intracellular signaling networks controlling numerous physiological and pathologic processes ranging from cell growth, stress-responses, aging, survival, to diabetes, autoimmunity and cancer. Understanding their roles in these processes, especially by focusing on their roles in the vascular system and immune responses are relevant for the mission of VBT. Specifically, our finding of fine interaction between MEKK3 and CCM2 and the critical role of MEKK3 in brain vascular integrity is highly relevant for the mission of VBT. 4. Publications: (Publications July 1, 2013– June 30, 2014) Kaur S, Sassano A, Majchrzak-Kita B, Baker DP, Su B, Fish EN, Platanias LC. Regulatory effects of

mTORC2 complexes in type I IFN signaling and in the generation of IFN responses. Proc Natl Acad Sci U S A. 2012 May 15;109(20):7723-8. Epub 2012 May 1. PMID:22550181.Functional analysis of the distal region of the third intracellular loop of PROKR2. Zhou XT, Chen DN, Xie ZQ, Peng Z, Xia KD, Liu HD, Liu W, Su B, Li JD. Biochem Biophys Res Commun. 2013 Sep 13;439(1):12-7.

Liu P, Gan W, Inuzuka H, Lazorchak AS, Gao D, Arojo O, Liu D, Wan L, Zhai B, Yu Y, Yuan M, Kim BM, Shaik S, Menon S, Gygi SP, Lee TH, Asara JM, Manning BD, Blenis J, Su B, Wei W. Sin1 phosphorylation impairs mTORC2 complex integrity and inhibits downstream Akt signalling to suppress tumorigenesis. Nat Cell Biol. 2013 Nov;15(11):1340-50. doi: 10.1038/ncb2860. Epub 2013 Oct 27. PMID:24161930.

Kaur S, Kroczynska B, Sharma B, Sassano A, Arslan AD, Majchrzak-Kita B, Stein BL, McMahon B, Altman JK, Su B, Calogero RA, Fish EN, Platanias LC. Critical roles for Rictor/Sin1 complexes in IFN-dependent gene transcription and generation of antiproliferative responses. J Biol Chem. 2014 Jan 27. [Epub ahead of print]. PMID:24469448.


54

Yajaira Suárez, PhD Assistant Professor of Comparative Medicine and Pathology.

1. Overall Goal(s) of the Research Program of the Laboratory: The Suárez laboratory studies the contribution of non-coding RNAs, including microRNAs, to the regulation of endothelial cell and macrophage functions. Both cell types play major role in controlling both angiogenic and inflammatory responses and the interplay between these two cell types has been shown to be critical for several pathophysiological conditions like atherosclerosis, cancer (tumor growth), adipose tissue expansion and wound healing, among others.

2. Specific Research Accomplishments in the last 12 months: We have described a novel role of miR-149 in controlling the steady state of FGF2-induced responses through the regulation of the GPC1-FGFR1 binary complex. Our findings show that FGF2 stimulates the expression of intronic miR-149 independently of its host gene to provide a fine-tuned regulation of GPC1 and FGFR1, co-receptor and receptor for FGF2, respectively, therefore assuring the steady state of FGF2-induced responses in ECs. We have published our findings related to the role of miR-155 in wound healing which indicates that miR-155 negatively regulates wound closure and that its therapeutic inhibition may be a promising treatment for patients with impaired dermal wound healing.

3. Significance of Key Findings Relevant for the Mission of VBT: Our research contribute to the understanding of the complex network involving miRNAs and their targets, leading to a coordinate pattern of gene expression that regulates the cellular and molecular mechanisms that control endothelial and macrophage functions. Our research can form the basis of new therapies for inflammation and angiogenesis. 4. Publications: (Primary Research Publications July 1, 2013– June 30, 2014) Goedeke L, Salerno A, Ramírez CM, Guo L, Allen RM, Yin X, Langley SR, Esau C, Wanschel A, Fisher

EA, Suárez Y, Baldán A, Mayr M, Fernández-Hernando C. Long-term therapeutic silencing of miR-33 increases circulating triglyceride levels and hepatic lipid accumulation in mice. EMBO Mol Med. 2014 Jul 18;6(9):1133-41. PubMed PMID: 25038053.

van Solingen C, Araldi E, Chamorro-Jorganes A, Fernández-Hernando C, Suárez Y. Improved repair of dermal wounds in mice lacking microRNA-155. J Cell Mol Med. 2014 Jun;18(6):1104-12. PubMed PMID: 24636235; PubMed Central PMCID: PMC4112003.

Chamorro-Jorganes A, Araldi E, Rotllan N, Cirera-Salinas D, Suárez Y. Autoregulation of glypican-1 by intronic microRNA-149 fine tunes the angiogenic response to FGF2 in human endothelial cells. J Cell Sci. 2014 Mar 15;127(Pt 6):1169-78. PubMed PMID: 24463821; PubMed Central PMCID: PMC3953812.

Dávalos A, Suárez Y. MiRNA-based therapy: from bench to bedside. Pharmacol Res. 2013 Sep;75:1-2. PubMed PMID: 23827159.

Chamorro-Jorganes A, Araldi E, Suárez Y. MicroRNAs as pharmacological targets in endothelial cell function and dysfunction. Pharmacol Res. 2013 Sep;75:15-27. PubMed PMID: 23603154; PubMed Central PMCID: PMC3752325.

Ramírez CM, Goedeke L, Rotllan N, Yoon JH, Cirera-Salinas D, Mattison JA, Suárez Y, de Cabo R, Gorospe M, Fernández-Hernando C. MicroRNA 33 regulates glucose metabolism. Mol Cell Biol. 2013 Aug;33(15):2891-902. PubMed PMID: 23716591; PubMed Central PMCID: PMC3719675.

Ramírez CM, Rotllan N, Vlassov AV, Dávalos A, Li M, Goedeke L, Aranda JF, Cirera-Salinas D, Araldi E, Salerno A, Wanschel A, Zavadil J, Castrillo A, Kim J, Suárez Y, Fernández-Hernando C. Control of cholesterol metabolism and plasma high-density lipoprotein levels by microRNA-144. Circ Res. 2013 Jun 7;112(12):1592-601. PubMed PMID: 23519695; PubMed Central PMCID: PMC3929583.


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Goedeke L, Vales-Lara FM, Fenstermaker M, Cirera-Salinas D, Chamorro-Jorganes A, Ramírez CM,

Mattison JA, de Cabo R, Suárez Y, Fernández-Hernando C. A regulatory role for microRNA 33* in controlling lipid metabolism gene expression. Mol Cell Biol. 2013 Jun;33(11):2339-52. PubMed PMID: 23547260; PubMed Central PMCID: PMC3648071


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George Tellides, M.D., Ph.D. Professor of Surgery (Section of Cardiac Surgery) and of Investigative Medicine; Chief of Cardiothoracic Surgery, Veterans Affairs Medical Center 1. Overall Goal(s) of the Research Program of the Laboratory: Vascular biology; Immunology; Transplantation 2. Specific Research Accomplishments in the last 12 months: Determining the role of TGF-b in homeostasis of the aorta. 3. Significance of Key Findings Relevant for the Mission of VBT: Collaborations with Pober lab on mechanisms of graft arteriosclerosis, with Simons lab on mechanisms of endothelial to mesenchymal transformation, and with Humphrey/Schwartz labs on the role of mechanosensing in aortic disease. 4. Publications: (Publications July 1, 2013– June 30, 2014) Kondo Y, Jadlowiec CC, Muto A, Yi T, Protack C, Collins MJ, Tellides G, Sessa WC, Dardik A. The

Nogo-B-PirB axis controls macrophage-mediated vascular remodeling. PLoS One. 2013 Nov 20;8(11):e81019. doi:10.1371/journal.pone.0081019. eCollection 2013. PubMed PMID: 24278366; PubMed Central PMCID: PMC3835671.

Liu R, Jin Y, Tang WH, Qin L, Zhang X, Tellides G, Hwa J, Yu J, Martin KA. Ten-eleven translocation-2 (TET2) is a master regulator of smooth muscle cell plasticity. Circulation. 2013 Oct 29;128(18):2047-57. doi:10.1161/CIRCULATIONAHA.113.002887. Epub 2013 Sep 27. PubMed PMID: 24077167;PubMed Central PMCID: PMC3899790.

Jane-Wit D, Manes TD, Yi T, Qin L, Clark P, Kirkiles-Smith NC, Abrahimi P, Devalliere J, Moeckel G, Kulkarni S, Tellides G, Pober JS. Alloantibody and complement promote T cell-mediated cardiac allograft vasculopathy through noncanonical nuclear factor-κB signaling in endothelial cells. Circulation. 2013 Dec 3;128(23):2504-16. doi: 10.1161 /CIRCULATIONAHA 113.002972. Epub 2013 Sep 17. PubMed PMID: 24045046; PubMed Central PMCID: PMC3885874.

Roccabianca S, Figueroa CA, Tellides G, Humphrey JD. Quantification of regional differences in aortic stiffness in the aging human. J Mech Behav Biomed Mater. 2014 Jan;29:618-34. doi: 10.1016/j.jmbbm.2013.01.026. Epub 2013 Feb 9.PubMed PMID: 23499251; PubMed Central PMCID: PMC3842391.

Qin L, Huang Q, Zhang H, Liu R, Tellides G, Min W, Yu L. SOCS1 prevents graft arteriosclerosis by preserving endothelial cell function. J Am Coll Cardiol. 2014 Jan 7-14;63(1):21-9. doi: 10.1016/j.jacc.2013.08.694. Epub 2013 Aug 28. PubMed PMID: 23994402; PubMed Central PMCID: PMC3932325.

Wang C, Qin L, Manes TD, Kirkiles-Smith NC, Tellides G, Pober JS. Rapamycin antagonizes TNF induction of VCAM-1 on endothelial cells by inhibiting mTORC2. J Exp Med. 2014 Mar 10;211(3):395-404. doi: 10.1084/jem.20131125. Epub 2014 Feb 10.PubMed PMID: 24516119; PubMed Central PMCID: PMC3949571.

Chen PY, Qin L, Zhuang ZW, Tellides G, Lax I, Schlessinger J, Simons M. The docking protein FRS2α is a critical regulator of VEGF receptors signaling. Proc Natl Acad Sci U S A. 2014 Apr 15;111(15):5514-9. doi: 10.1073/pnas.1404545111. Epub 2014 Apr 2. PubMed PMID: 24706887; PubMed Central PMCID: PMC3992672.

Humphrey JD, Milewicz DM, Tellides G, Schwartz MA. Cell biology. Dysfunctional mechanosensing in aneurysms. Science. 2014 May 2;344(6183):477-9. doi: 10.1126/ science. 1253026. PubMed PMID: 24786066.

Pober JS, Jane-wit D, Qin L, Tellides G. Interacting mechanisms in the pathogenesis of cardiac allograft vasculopathy. Arterioscler Thromb Vasc Biol. 2014 Aug;34(8):1609-14. doi: 10.1161/ATVBAHA.114.302818. Epub 2014 Jun 5. Review. PubMed PMID: 24903097


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Liu R, Jin Y, Tang W, Qin L, Zhang X, Tellides G, Hwa J, Yu J, Martin KA. Response to Letter

Regarding Article, "Ten-Eleven Translocation-2 (TET2) Is a Master Regulator of Smooth Muscle Cell Plasticity". Circulation. 2014 Aug 19;130(8):e72. doi: 10.1161/CIRCULATIONAHA . 114.010272. PubMed PMID: 25135133.


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Daniela Tirziu, Ph.D. Research Scientist in Medicine (Cardiology) 1. Overall Goal(s) of the Research Program of the Laboratory: My laboratory investigates the growth regulatory mechanisms generated by endothelium-cardiomyocyte crosstalk that control the cardiac size and function. Our ongoing study on myocardial angiogenesis and hypertrophy has identified a novel molecular mechanism depended on endothelial-nitric oxide release that coordinates angiogenesis with cardiomyocyte growth, published in J Clin Invest (2013). This reveals a new and unexplored role played by the vascular endothelium in regulating adult organ growth and size that is highly relevant to understanding the potential role of therapeutic angiogenesis during cardiac hypertrophy and failure. 2. Specific Research Accomplishments in the last 12 months: Based on our recent work, which showed that myocardial angiogenesis promotes myocardial hypertrophy through the action of endothelial-derived NO to induce cardiomyocyte RGS4 degradation with de-repression of Gβγ and activation of PI3Kγ/Akt/mTORC1 pathway, we focused on the role of microRNAs in endothelium to cardiomyocyte crosstalk. Our recent studies have identified that the increase of miR-182 in cardiomyocytes plays an important role in mediating the hypertrophic response subsequent to angiogenesis in heart. Moreover, we have found that miR-182 fine tunes the hypertrophic response propagated through NO/PI3Kγ/Akt/mTORC1 pathway by targeting PTEN induced putative kinase 1 (Pink1), adenylate cyclase type 6 (Adcy6), branched chain aminotransferase 2 (Bcat2) and forkhead box O3 (Foxo3) to potentially increase the anabolic effect of branched chain aminoacids (BCAA) on protein synthesis and cellular mass (manuscript under review). 3. Significance of Key Findings Relevant for the Mission of VBT: We continue to explore the growth signaling pathways involved in endothelium-cardiomyocyte crosstalk and to determine the therapeutic potential of vascular endothelium-derived paracrine factors in clinical settings relevant to cardiovascular disease and heart failure. Better understanding of the biological processes generated by cell-cell communications will provide mechanist insights and potential new therapeutic avenues aimed at treating cardiovascular disease. 4. Publications: (Publications July 1, 2013– June 30, 2014) Li N, Jaba IM, Han J, Larrivée B, Tirziu D. mir-182 modulates myocardial hypertrophic response induced

by angiogenesis in heart. Circulation. 2013; 128:A15179 (abstract) Stacy MR, Yu da Y, Maxfield MW, Jaba IM, Jozwik BP, Zhuang ZW, Lin BA, Hawley CL, Caracciolo CM,

Pal P, Tirziu D, Sampath S and Sinusas AJ: Multimodality imaging approach for serial assessment of regional changes in lower extremity arteriogenesis and tissue perfusion in a porcine model of peripheral arterial disease. Circ Cardiovasc Imaging. 2014 Jan;7 (1) :92-9. Epub 2013 Oct 29. PMID: 24170237

Sakurai T, Lanahan A, Woolls MJ, Li N, Tirziu D and Murakami M: Live cell imaging of primary rat neonatal cardiomyocytes following adenoviral and lentiviral transduction using confocal spinning disk microscopy. J Vis Exp. 2014 Jun 24; (88) :e51666. Epub 2014 Jun 24. PMID: 24998400


59

Agnès Vignery, DDS, PhD Senior Research Scientist, Orthopaedics and Cell Biology

1. Overall Goal(s) of the Research Program of the Laboratory: Research in our laboratory focuses on two lines of investigation, the commonality of which is bone metabolism, which is at the cross-road of the vascular, immune, endocrine and nervous systems. The first line of investigation regards the differentiation of osteoclasts in bone, and/or giant cells in chronic inflammatory reactions and cancer. Although osteoclasts and giant cells play a central role in these diseases, the molecular mechanisms that are responsible for their development remains poorly understood. Research in our lab focuses on the molecular mechanisms of fusion of their precursor cells, which belong to the monocyte-macrophage lineage. The second line of investigation focuses on the targeted engineering of new bone to specific sites of the skeleton that are at risk for fracture, or do no repair well from fracture. 2. Specific Research Accomplishments in the last 12 months: Board member, The Journal of Immunology, extended till 2015 Keynote Speaker at The 7th International “Oral Life Bioscience” Symposium, Kyushu University, Fukuoka, Japan Invited Speaker at the Imperial College London, Imperial College London, Centre for Complement and Inflammation Research (CCIR), Hammersmith Hospital Invited Speaker and Session Chair at the Embo Workshop on Cell-Cell Fusion, Ein Gedi, Israel Invited Speaker, Department of Orthopaedics, University of Illinois at Chicago, Chicago, IL Macrophage Café (December 2012-present) Initiator and Organizer Monthly Luncheon, Yale School of Medicine, which is designed to open up a forum, the goals of which are:

Discuss the roles that macrophages play in tissue/organ homeostasis and in diseases, and challenge established dogmas about macrophage functions.

Broaden our understanding of how macrophages interact with cells, such as endothelial cells, nerves, mesenchymal cells, pathogens, etc.

Discuss the role played by macrophages in biological systems and in cancer 3. Significance of Key Findings Relevant for the Mission of VBT: The formation of intramembranous bone, in contrast with endochondral bone, requires extensive vascularization. Our finding that impressive quantities of new bone form in targeted sites of the skeleton as a result of marrow aspiration and daily treatment with parathyroid hormone has revealed an essential link between the formation of intramembranous bone and vascularization, which we are actively exploring. 4. Publications: (Publications July 1, 2013– June 30, 2014) Kang H, Kerloc'h A, Rotival M, Xu X, Zhang Q, D'Souza Z, Kim M, Scholz JC, Ko JH, Srivastava PK,

Genzen JR, Cui W, Aitman TJ, Game L, Melvin JE, Hanidu A, Dimock J, Zheng J, Souza D, Behera AK, Nabozny G, Cook HT, Bassett JH, Williams GR, Li J, Vignery A*, Petretto E, Behmoaras J. Kcnn4 is a regulator of macrophage multinucleation in bone homeostasis and inflammatory disease. Cell Rep. 2014 Aug 21;8(4):1210-24. doi: 10.1016/j.celrep.2014.07.032. Epub 2014 Aug 14. PubMed PMID: 25131209 Vignery A* co-senior author


60

Dianqing (Dan) Wu, PhD

Professor of Pharmacology

1. Overall Goal(s) of the Research Program of the Laboratory: The overall objective of our research activities is to understand the mechanisms and functions of signal transduction activated by chemoattractants and Wnts. 2. Specific Research Accomplishments in the last 12 months: The key research accomplishment includes: 1) Uncovering a previously unknown mechanism by which the STK24 and complex regulates ligand-gated exocytosis and acquiring preliminary evidence for involvement of this mechanism in pathogenesis of CCM disease in collaboration with Wang Min’s lab. 2) Investigation of the role of SRF in neutrophil migration in collaboration with Stephanie Halene’s lab. 3) Characterization of a mechanism for HGF to crosstalk with Wnt signalling in collaboration with Lloyd Cantley’s lab. 4) Elucidation of the mechanisms by which different Raf kinases regulate cell migration via stimulation of E3 ligase RFFL. 3. Publications (Publications July 1, 2011– June 30, 2012): Gan, X., Wang, C., Patel, M., Kreutz, B., Zhou, M., Kozasa, T., and Wu, D., Different Raf protein kinases

mediate different signaling pathways to stimulate E3 ligase RFFL gene expression in cell migration regulation. J Biol Chem, 2013. 288: 33978-84. PMC: 3837137

Zhang, Y., Tang, W., Zhang, H., Niu, X., Xu, Y., Zhang, J., Gao, K., Pan, W., Boggon, T.J., Toomre, D., Min, W., and Wu, D., A network of interactions enables CCM3 and STK24 to coordinate UNC13D-driven vesicle exocytosis in neutrophils. Dev Cell, 2013. 27: 215-26. PMC: 3834565

Zhang, Y., Tang, W., Zhang, H., Niu, X., Xu, Y., Zhang, J., Gao, K., Pan, W., Boggon, T.J., Toomre, D., Min, W., and Wu, D., A network of interactions enables CCM3 and STK24 to coordinate UNC13D-driven vesicle exocytosis in neutrophils. Dev Cell, 2013. 27: 215-26. PMC: 3834565

Taylor, A., Tang, W., Bruscia, E.M., Zhang, P.X., Lin, A., Gaines, P., Wu, D., and Halene, S., SRF is required for neutrophil migration in response to inflammation. Blood, 2014. 123: 3027-36. PMC: 4014845


61

Lawrence H. Young, M.D. Professor of Medicine (Cardiology) and of Cellular and Molecular Physiology; Vice-Chairman Department of Medicine 1. Overall Goal(s) of the Research Program of the Laboratory: Key areas of our research are the molecular & cellular mechanisms regulating heart (metabolism in cv disease), autocrine-paracrine pathways (modulating cardiac cell signaling and function), and metabolic signaling pathways in atrial fibrillation. Processes under investigation include: regulation of heart metabolism/function; AMPK protection against solid organ ischemia; JNK pathway in reperfusion injury; cardiac autocrine/paracrine factors; MIF CD74 signaling; novel myocyte autocrine proteins; and LKB1 role in cardiac growth and remodeling. 2. Specific Research Accomplishments in the last 12 months: Our work has defined a novel autocrine-paracrine pathway in the heart that is critical to the myocardial response to ischemia-reperfusion. D-dopachrome tautomerase is expressed in cardiac myocytes and the coronary vasculature, is secreted during ischemia and activates metabolic signaling in the heart. We also have shown that urocortin mediates its cardioprotective actions via the AMP-activated protein kinase signaling pathway. Finally, in collaboration with Dr. Tirziu we have elucidated the paracrine effects of endothelium derived nitric oxide in stimulating myocardial hypertrophy during angiogenesis. 3. Significance of Key Findings Relevant for the Mission of VBT: Our work continues to advance the understanding of the interaction between the cross-talk between the vasculature and myocardium. 4. Publications: (Publications July 1, 2013– June 30, 2014) Jaba IM, Zhuang ZW, Jiang Y, Martin K, Li N, Sinusas A, Papademetris X, Simons M, Sessa W, Young

LH, Tirziu D. Nitric oxide-regulated RGS4 loss of function coordinates vasculature and cardiomyocyte growth. J Clin Invest. 2013;123:1719-1731.

Li J, Kim A, Hu X, Miller EJ, Wu X, Qi D, Xiao L, Sherwin R, Young LH. Urocortin 2 autocrine-paracrine and pharmacologic effects to activate AMP-activated protein kinase in the heart. PNAS. 2013;110:16133-16138.

Qi D, Atsina K, Qu L, Piecychna M, Leng L, Bucala R, Young LH. The vestigial enzyme D-dopachrome tautomerase protects the heart against ischemic injury. J Clin Invest 2014; 124:3540-3550.


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Jun Yu, MD

Research Scientist, Internal Medicine - Cardiology

1. Overall Goal(s) of the Research Program of the Laboratory: The primary research goals in my laboratory are to understand the molecular control of vascular remodelling and atherogenesis in response to arterial injury. We have intensively used mouse genetic, cell biological and biochemical approaches to achieve these goals. Currently, our major efforts are directed at:

a) Endoplasmic Reticulum (ER) stress-induced ER remodeling in regulation of therosclerotic plaque progression and instability.

b) The role of primary cilia in shear induced vascular remodelling and atherogenesis. c) Defining the role of Reticulon-4B in platelet activation and apoptosis in diabetes.

2. Specific Research Accomplishments in the last 12 months: In the past year, we have made significant progresses in several research areas: (a) In the study of examine the role of reticulon family protein Nogo-B in macrophage function and atherogenesis, we have found that loss of Nogo-B accelerated plaque formation which leading to severe coronary lesions and led to morphologically more advanced lesion in vivo. Macrophages that lack of Nogo-B are more susceptible to modified LDL (mLDL) induced foam cell formation and free cholesterol induced apoptosis. Our research discovered, for the first time, that mLDL upregulates Nogo-B and scavenger receptor A (SR-A) through JNK-c-Jun/AP-1 pathway. Nogo-B in turn negatively regulates SR-A to limit mLDL uptake by interacting with MEKK2/3. Nogo-B is necessary for regulating ER-mitochondria complex and thus control macrophage apoptosis. (b) In a collaboration with Drs. Martin Schwartz and Stefan Somlo, we are uncovering a new potential functions of polycystins (PC) -1 and -2 in regulating shear stress induced inflammatory responses in endothelial cells in vitro. We are now in the process of investigating the contribution of PC -1 and -2 in atherogenesis. (c) In a collaboration with Dr. John Hwa, we have identified Nogo-B as a key regulator of platelets activation and apoptosis in type II diabetes. (d) We have been collaborating continuously with other research groups in the School of Medicine and have contributed to several publications during the past year as reflected below.

3. Significance of Key Findings Relevant for the Mission of VBT: Our findings are significant and in line with the mission of VBT program that uncover new signal pathway in regulating inflammation and vascular remodeling.

4. Publications: (2013– 2014) Qiu C, Yu J. The Function of Polycystin-1 and Polycystin-2 in Cardiovascular System. J Cardiovasc Dis

Diagn. 2013, 1:110. doi: 10.4172/jcdd.1000110. Liu R, Jin Y, Tang W, Qin L, Zhang X, Tellides G, Hwa J, Yu J, Martin KA. TET2 is a master regulator of

smooth muscle cell plasticity. Circulation. 2013; 128(18):2047-57. PMID: 24077167 Di Lorenzo A, Lin MI, Murata T, Landskroner-Eiger S, Schleicher M, Kothiya M, Iwakiri Y, Yu J, Huang

PL, Sessa WC. eNOS derived nitric oxide regulates endothelial barrier function via VE cadherin and Rho GTPases. J Cell Sci. 2013; 126(Pt 24):5541-52. PMCID: PMC3860306

Qiu C, Jozsef L, Yu B, Yu J. Saponin monomer 13 of dwarf lilyturf tuber (DT-13) protects serum withdrawal-induced apoptosis through PI3K/Akt in HUVEC. BBRC. 2014; 443(1):74-9. PMID: 24269237

Mehra VC, Jackson E, Zhang XM, Jiang XC, Dobrucki LW, Yu J, Bernatchez P, Sinusas


APPENDIX 1

The Thirteenth Annual VBT Retreat

The Thirteenth Annual Retreat of the

Vascular Biology and Therapeutics Program held in conjunction with Cardiovascular Medicine

Friday, October 18, 2013

The Grace Murray Hopper Auditorium, Building @-B25, West Campus

8:00-8:30

Registration -Continental Breakfast

Session I 8:30-12:00

Session Chair – Jordan Pober, MD, PhD

8:30-9:00 John Hwa, Ph.D. "Hyperglycemia and platelet mitochondrial dysfunction" Questions

9:00-9:30 Carlos Fernandez-Hernando, PhD “Identification of novel genes involved in macrophage foam cell formation: Role of angiopoetin-like-4 (Angptl4) in the progression of atherosclerosis” Questions

9:30-10:00

Angelica Gonzalez, PhD "Advanced Models of Microvasculature: Composite Cellular and Matrix Structures Regulate Leukocyte Recruitment" Questions

10:00-10:20 Break

10:20:10:50 Hyung Chun, MD “GPCR crosstalk in vascular maturation” Questions

10:50-11:00 William Sessa, PhD Introduction of Keynote Speaker

11:00 – 12:00 Keynote Address Ralf H. Adams, PhD, Professor, University of Muenster, Director Max Planck Institute for Molecular Biomedicine “New Branches on the vascular tree: vessel heterogeneity and functional specialization” Questions

12:00 – 1:30 Lunch & Poster Session

SESSION II 1:30 – 4:20

Session Chair – Anne Eichman, PhD

1:30 – 2:00 Albert Sinusas, MD “Imaging of angiogenesis and arteriogenesis: Mouse to man” Questions

2:00 – 2:30 Jaime Grutzendler, MD “Mechanisms of postnatal cerebral microvascular plasticity” Questions

2:30 - 3:00 Stefania Nicoli, PhD The diverse functions of microRNAs in animal development Questions

3:00-3:20 Break

3:20-3:50 Kathleen Martin, PhD “TET2 is a novel master regulator of vascular smooth muscle phenotype” Questions

3:50 – 4:20 Yajaira Suarez, PhD

“Integration of Macrophage Inflammatory Responses and Cholesterol Metabolism” Questions

4:20-4:30 Announcement of Poster Contest Winners

Vascular Biology & Therapeutics

VASCULAR BIOLOGY AND `THERAPEUTICS ANNUAL REPORT 2013 – 2014

APPENDIX 2

The Twelfth Annual Yale – Cambridge Collaboration Meeting

Saturday, September 7 12:15 - Pickup at JFK, Hys Car Service – Confirmation #137269 When everyone is together, call T: 800-255-LIMO (5466) or T: 203-934-6331 5:00 – 6:00 Wine & Cheese in New Haven Hotel Lobby 6:00 Buffet Dinner, New Haven Hotel Dining Room – 2nd Floor Sunday, September 8 7:45 – 8:30 Breakfast - Amistad

8:30 – 8:40 Opening Remarks – John Bradley & Jordan Pober

8:40 – 2:30 Session 1 – Cardiovascular

Chairs – William Sessa and Patrick Maxwell

8:40 – 9:00 John Hwa – “Hyperglycemia, aldose reductase and platelet dysfunction”

9:00 – 9:20 Martin Bennett –'Mitochondria in atherosclerosis'

9:20 – 9:40 Wang Min–“TNFR2 signaling in cardiac progenitor”

9:40 – 10:00 Break

10:00 – 10:20 Paul Upton – “BMP9 regulates junctional proteins in endothelial cells: implications for vascular dysplasias”

10:20 – 10:40 Alan Dardik- “Venous adaptation to the arterial circulation”

10:40 – 11:00

Roderick Llewelyn – “Epigenomics of cardiac hypertrophy”

11:00 – 11:20 George Tellides – “mTOR Inhibition for Aortic Disease”

11:20 – 12:00 Lunch

12:00 – 12:20 John Waters – “3-D modelling of glomerulosclerosis”

12:20 – 12:40 Mehran Sadeghi –“Imaging protease activation in vascular and valvular disease”

12:40 – 1:00 Rameen Shakur – “Genome Editing and Modelling Sudden Cardiac death in the Cardiomyopathies

1:00 – 1:20 Marty Kluger - “TNF Induces Three Sequential Changes in Human Dermal Microvascular Endothelial Cell Barriers: An in vitro Model of Inflammatory Capillary Leak”

1:20 – 1:40 Ziad Mallatt – “B cell/monocyte interactions in acute myocardial infarction”

1:40 – 2:00 Carlos Fernandez-Hernando–“microRNA regulation of lipid metabolism”

2:00 – 2:20 William Sessa – “Endothelial control of lipid metabolism”

2:30 Bus will pick-up Cambridge Attendees at Amistad to outdoor party in Guilford

Monday, September 8, 2013 7:45 – 8:30 Continental Breakfast – Amistad Building

8:30 – 12:00 Session 2 –Neuroscience/Immunology

Chairs – David Hafler

8:30 – 8:50 Joanne Jones –“Alemtuzumab - "fixing" multiple sclerosis and causing autoimmunity”

8:50 – 9:10 Kevin O’Connor – “Related B cell clones populate the brain and periphery of patients with multiple sclerosis”

9:10 – 9:30 Doug Fearon – “A stromal cell lineage with roles in tumor immune suppression,cachexia, hematopoiesis, mesenchylmal organ development, and growth”

9:30 – 9:50 Eric Meffre – "The thin line between autoimmunity and immunodeficiency"

9:50 – 10:20 BREAK

10:20 – 10:40 Murat Gunel – TITLE PENDING

10:40 – 11:00 David Hafler – “ENCODE decoding of MS Genetic Architecture”

11:00 – 11:20 Su Metcalfe – "Developing a platform for Personalised Medicine in Parkinson's Disease"

11:20 – 11:40 Mark Evans – “Brain glucokinase and glucose homeostasis”

11:40 – 1:00 Lunch 1:00 – 5:30 Session 3 – Immunology

Chairs – Jordan Pober and Sir Keith Peters

1:00 – 1:20 Arthur Kaser – “The endoplasmic reticulum inflames the intestine”

1:20 – 1:40 Kevan Herold – “Analysis of antigen specific T cells in individuals at risk for development of Type 1 diabetes”

1:40 – 2:00 Frank Waldon-Lynch – “Preclinical and clinical studies of ultra low dose IL-2 in type 1 diabetes”

2:00 – 2:20 Stephanie Eisenbarth – “The unique and divided roles of splenic dendritic cell subsets in T cell activation”

2:20 – 2:40 Larry Peterson– “A path towards improving IL-2 for treating autoimmunity”

2:40 – 3:00 João P. Pereira – “Movement of hematopoietic cells during differentiation in bone marrow”

3:00 – 3:30 BREAK

3:30 – 3:50 Paul Lehner – “Genetic and Functional Proteomic Approaches to Analysing Viral Evasion Pathways”

3:50 – 4:10 Phil Askenase – “Nano Immuno Genetic Therapy: Antigen-Specific Suppressor T Cell Exosomes Deliver Inhibitory miRNA-150”

4:10 – 4:30 Andres Floto – “A Spaetzle-like role for nerve growth factor in vertebrate immunity to Staphylococcus aureus”

4:30 – 4:50 Sanjay Kulkarni – “Eculizumab Therapy for Chronic Antibody Mediated Injury in Kidney Transplantation”

4:50 – 5:10 James Nathan – “New insights into decoding ubiquitin signals”

5:10 – 5:30 Jordan Pober - "Inhibition of Endothelial mTOR Signaling Reduces Inflammation”

5:30 – 5:45 OPEN

5:45 – Bus pick-up at Amistad to Anthony’s Ocean View

6:00 Anthony’s Ocean View

Tuesday, September 10. 2013 7:45 – 8:30 Continental Breakfast – Amistad Building

8:30 – 12:00 Session 4 – Stem Cell

Chairs – Haifan Lin and Ludovic Vallier

8:30 – 8:50 Rafia Al-Lamki / John Bradley – “'Modulation of stem and differentiated cells in renal cell carcinoma by tumour necrosis factor”

8:50 – 9:10 Jun Lu –An Extensive Network of TET2-Targeting microRNAs Regulates Malignant Hematopoiesis"

9:10 – 9:50 Willem Ouwehand - Large scale sequencing initiatives in the UK Augusto Rendon - NGS in the discovery and diagnosis of bleeding and platelet disorders

9:50 – 10:10 Diane Krause - Role of the actin cytoskeleton in maturation of megakaryocytes”

10:10 – 10:40 BREAK

10:40 – 11:00 Stefano Pluchino – “'How neural stem cells speak with the host immune

11:00 – 11:20 Sanja Sinha – “Modelling Human Vascular Development and Disease using Pluripotent Stem Cells”

11:20 – 11:35 Shangqin Guo – "Visualizing the birth of induced pluripotency"

11:40 – 12:00 Ludovic Vallier – “Cell cycle rules stem cell fate”

12:00 – 12:20 Valentina Greco - "Stem cell fate and function in the hair follicle stem cell niche by live imaging"

12:30 Lunch 2:30 Bus pick-up Amistad

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