SDR™ and SRG™ SID Precision Toxicology™ & Efficacy ... · Precision Toxicology™ & Efficacy:...
Transcript of SDR™ and SRG™ SID Precision Toxicology™ & Efficacy ... · Precision Toxicology™ & Efficacy:...
SDR™ and SRG™ SCID Rats
Cancer Xenografts
Liver & Immune system Humanization
Hera [email protected]
Precision Toxicology™ & EfficacyCRO Services
About Hera BioLabs
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2015 2016 2017 2017/18 plan
Hera spun-out of Transposagen
& licenses IP for gene editing
technology; development of
SCID rats begins; awarded phase
II SBIR grant
Completion of a 10,000 ft2
facility; Scientific team assembled
with in vitro & in vivo efficacy &
toxicity capabilities
Introduction of SDR™ & SRG™
SCID rats and efficacy
services; Engineered HepG2
and MDCK cells; in vivo
toxicity studies and
humanized liver mice; custom
gene editing, breeding and
screening services in mouse
and rat
Humanization of the liver &
immune system of SRG™ rats for
toxicity and immuno-oncology
services
Precision Toxicology™ & Efficacy: utilizing precisely gene-edited models such as SCID rats, humanized rodents and engineered cell lines for producing more rapid, consistent and clinically-relevant data
Gene editing technology & capabilities
Hera’s gene editing tool box for product development
Custom gene editing, phenotyping and screening services in cells, rats & mice available3
Gene Editing Tool Box
Site-Specific Nucleases
TALEN CRISPR/Cas9
piggyBac™
Hyperactive piggyBac™
Excision-only piggyBac™
Bioinformatics
• Stable, transgenic integrations
• Genetic screens
• Footprint-free™ gene editing• Gene editing enhancements
• Gene knock-in & knockout
• Better project design• Off-target analysis
Hera’s products & services
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Disease Modeling
Links for specific product and service information above
Why use SCID rats for xenograft efficacy studies?
Key differentiator
In mice, many tumor models do not engraft or have very low and inconsistent take rates and growth kinetics, thus conducting a robust efficacy study is not feasible. There is an unmet need for better xenograft models. Hera’s SCID rats (SDR™ & SRG™ Rats) have demonstrated better take rates and growth kinetics than mouse models, providing for more rapid and consistent data.
Additional benefits
Better and more human-like PK properties for most compounds compared to mice.
The rat’s size is appropriate for multiple dosing, particularly by routes other than oral, and for collection of multiple blood samples, which enable investigators to assess efficacy and systemic toxicity using fewer animals over longer time periods.
In xenograft studies, rats can grow larger (more human-sized) tumors and produce larger samples for analysis.
More precise surgical manipulations and better in-vivo imaging of metastatic tumors are possible.
The generally slower metabolic rate (resulting in lower dosages) and the use of fewer animals mitigates the increased amounts of test article that might be expected when switching from the mouse to the rat.
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Hera SCID Rat Models
SDR™ Rats (SD-Rag2 KO):
Knockout of the Rag2 (recombination activating gene 2) gene on the Sprague Dawley background (SD).
The knockout impairs V(D)J recombination and results in a loss of mature B cells and T cells.
SRG™ Rats (SD-Rag2-Il2rg KO):
Knockout of Rag2 and Il2rg (interleukin 2 receptor gamma chain) results in depletion of T cells, B cells, and NK cells.
The most immunodeficient rat model available.
Immunophenotyping slides are in the extra slides section at the end
SDR & SRG rats are covered by issued patents, see more details on slide 14
Non-Small Cell Lung Cancer (NSCLC) H358 Cell Line – KRAS mutant
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SDR rats had a 100% engraftment rate (compared to <20% in comparable mouse models) and showed better overall tumor growth kinetics.
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H358 HERA 1 million
H358 HERA 5 million
H358 HERA 10 million
H358 NGS 10 million
H358 Nude 10 million
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SCID mouse models
SDR rat
Human glioblastoma-derived tumors in Rag2 knockout (SDR) rats.
Tumors were measured twice weekly after inoculation of U87MG human glioblastoma cells subcutaneously. 6/6 Rag2 knockout animals developed measurable tumors by 27
days post-inoculation.
Confirmation of human tumors derived from subcutaneous injection of U87MG human glioblastoma cells
Immunohistochemistry (IHC) was done on tumors: brown anti-human mitochondria protein, peri-nuclear stain identified the tumors as human; hematoxylin (general nuclear stain) shown in blue/purple.
Rat Liver
Visible tumor in SDR rat
Additional tumor models xenografts confirmed in SCID rats
Model Description SCID rats Mice Notes
VCaP Hormone refractory prostate cancer cell line with high AR and PSA expression.
Yes, 100% take rate in pilot studies conducted by Hera in SRG rats
Very poor and inconsistent take rates, multiple mouse models and conditions tested by external established CRO
Mouse models are insufficient for efficacy studies due to low engraftment efficiency combined with highly variable tumor growth rates. Both problems are potentially alleviated in the SRG rat
Ovarian cancer PDX derived cell line (CDX)
Ovarian cell line derived from HGSOC patient. Applications include Parp inhibitor drug discovery
Yes, pilot studies conducted by Hera show engraftment in SDR rats
Yes, pilot studies conducted by Hera show engraftment in NSG mice from JAX
Head-to-head studies show better engraftment efficiency in SDR rat compared to NSG mice
Endometrial cancer PDX derived cell line
Endometrial cell line derived from serious endometrial patient. Applications include Parp inhibitor drug discovery
Yes, pilot studies conducted by Hera show engraftment in SDR rats
Yes, pilot studies conducted by Hera show engraftment in NSG mice from JAX
Head-to-head studies show faster engraftment in SDR rats compared to NSG mice, with 100% efficiency
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Hera’s xenograft efficacy services
Early adopters offer: no-risk pilot take rate & kinetics study for any cell or PDX model
Efficacy studies
SDR and SRG SCID rats are available off-the-shelf for studies at the clients facility
Studies conducted in commercially available mouse models or side-by-side species with Hera SCID rats for comparison studies (Hera has a license from JAX for NSG mice as well as has access to SCID mice from Taconic and CRL)
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Humanized rodent models in the pipeline
Humanized liver models allow for the detection of human specific liver toxicity and metabolism studies
Humanized liver mouse models have been successfully generated at Hera and services are available
Humanized liver rat models are under development and expected in late 2017
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Humanized immune system models are utilized for immuno-oncology studies & where the test article interacts with the immune system
Humanized immune system mice are commercially available for studies at Hera
Humanized immune system rats are under development and expected in 2018
Hera’s facility and scientific team
Vivarium featuring all 100% HEPA filtered, disposable IVC caging specifically designed for immunocompromised animals
Molecular and cell culture facility
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Dosing by multiple routes including i.p., oral and i.v.
ACLAM and AALAS certified staff
Digital data acquisition via StudyLog® systems software
Gene Editing & SCID Rat Intellectual Property
Hera has the freedom to operate through multiple licenses to issued and pending patents for CRISPR, piggyBac and TALEN gene editing technologies
Hera’s SCID rats are covered under issued patents claiming knockout rat phenotypes for immune system disorders, SCID and cancer. US Patent Numbers: 8,558,055; 9,314,005; 8,722,964
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Tseten Yeshi, Ph.D.
VP, R & DFormer Director of R&D at
Transposagen. An expert in genome editing with well-developed
scientific program management skills and experience.
Hera BioLabs Leadership
Chris Chengelis, Ph.D.,
DABTSenior Scientific AdvisorFormer CSO at WIL Research. 35
years+ experience in the preclinical toxicology industry, facility design,
study design and execution
Jack Crawford, M.S.
CEOFormerly directed the Sales,
Marketing, and Business Development Divisions at
Transposagen. Experience in product development, licensing,
technology and patent evaluation, and fundraising.
Fallon Noto, Ph.D.
Senior Scientist10+ years working with mice and
rats, expertise in rodent humanization, cell and tissue
transplantation, microsurgery, and ethical animal care.
Kamesh Ravi, Ph.D.
Senior Scientist10+ years of experience in
preclinical oncology, cancer xenograft models, tumor efficacy
studies and onco-nephrology.
Contact Us: [email protected] 859-414-0648 2277 Thunderstick Dr. #500 Lexington, KY 40505
Goutham Narla, M.D., Ph.D.SAB Member & ConsultantThe Pardee Gerstacker Professor of
Cancer Research and a Medical geneticist at Case Western Reserve
University. CSO and Scientific Founder of Dual Therapeutics, Inc.
Expertise in cancer genetics and xenograft and transgenic models of cancer with over 58 publications in
the field.
Immunophenotype of SDR™ rats
Analysis of immune cell populations in the SDR™ rat. A) SDR™ rat thymocytes contain fewer mature T cells (bottom panel), compared to a wild-type control (top panel). Insets show thymus with organ weight. The majority of thymocytes are CD4- and CD8-double negative in the SDR™ rat. B) The SDR™ spleen contains no mature B cells as demonstrated by lack of CD45R (B220)+/IgM+ cells (bottom panel) compared to wildtype spleen (top panel). C) SDR™ spleen has an increased NK cell population (bottom panel) compared to the wild-type (top panel).
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Wild Type
SDR
Immunophenotype of SRG™ rats
Analysis of immune populations in SRG™ rats. A) CD4+/CD8+ mature T cells are absent from SRG™ rat thymocytes (bottom panel), compared to a wild-type control (top panel). The lack of thymus tissue in the SRG™ rat results in a low recovery of thymocytes. B) The SRG™ spleen contains no mature B cells as demonstrated by lack of CD45R (B220)+/IgM+ cells (bottom panel), compared to WT spleen (top panel). C) The Il2rg knockout in the SRG™ rat results in a reduced NK cell population (bottom panel) compared to the SDR™ rat, which only has a Rag2 knockout (compare to figure 1, panel C). NK cells in the SRG™ rat are similar to or less than the amount of NK cells in the WT rat (top panel).
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Wild Type
SRG