SDR™ and SRG™ SCID Rats

Cancer Xenografts

Liver & Immune system Humanization

Hera BioLabsservices@herabiolabs.com

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.

0 20 40 600

1000

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5000

Treatment Days

Tu

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H358 HERA 1 million

H358 HERA 5 million

H358 HERA 10 million

H358 NGS 10 million

H358 Nude 10 million

Days post inoculation

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: services@herabiolabs.com 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.

Immunophenotyping Data

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