A new Rag2/Il2rg SCID rat Enabling cell line xenografts ... · Rag2/Il2rg double knockout. The most...

A new Rag2/Il2rg SCID rat

Enabling cell line xenografts

Accelerating PDX establishment

Humanizing the immune system

Tseten Yeshi, Ph.D.

VP R&D

services@herabiolabs.com

859-414-0648

About Hera BioLabs

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Novel Technologies

piggyBacTM

Cas-CLOVERTM

Proprietary Platforms

The SRG Rat

Engineered Cell Lines

Strong R&D Pipeline

Humanized Immune System

Humanized Liver

SRG

2015 20192016 2017 2018

Why Rat?

Better Data Quality – Translatability, Success

Better physiology, pharmacokinetics

Tolerability, toxicology, safety studies in rats

Larger blood volume: Serial blood draws from the same animal

Larger size: ease of surgical procedures, 10x tumor size

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Hera’s SRG Rat Model Rag2/Il2rg double knockout. The most immunodeficient commercially available rat model.

Knockout of the Rag2 (recombination activating gene 2) gene impairs V(D)J recombination and results in a loss of mature B cells and T cells.

The Il2rg (interleukin 2 receptor gamma chain) gene knockout leads to a lack of cytokine signaling, resulting defective lymphoid development.

The combined mutations result in a loss of mature B, T, and NK cells.

The background strain is Sprague Dawley.

The SRG platform

Immunophenotype of SRG rats

Analysis of immune populations in SRG rats. A) Athymic, CD4+/CD8+ mature T cells are absent. B) The spleen contains no mature B cells as demonstrated by lack of CD45R (B220)+/IgM+ cells. C) The Il2rg knockout results in a reduced NK cell population in the spleen.

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Wild type rat

SRG rat

NK Cells in SRG vs. Nude Rats

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Comparison between Nude rats and the SRG rat show a severally reduced number of natural killer (NK) cells in the circulating blood.

SRG ratNude rat

Outperforming the mouse for xenografts

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Feature Mouse OncoRat SRG

Engraftment Variable High

Growth profile Slower, variable Faster, uniform

Tumor size 1,000mm3 10,000+ mm3

Physiology Not ideal More human-like

Cell line xenografts

VCaP

OV81.2HCT116

VCaP exhibits many clinical characteristics and is ideal for AR resistance

Very difficult cell line to grow in mice: <20% take rate, highly variable growth profile

>90% tumor uptake in OncoRat SRG, more uniform growth profile

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Individual tumor volume in OncoRat SRG

Enabling Case Study: VCaP in OncoRat SRG

Serum PSA levels correlative to tumor volume

Case Study: VCaP in OncoRat SRG

Highly improved engraftment efficiency and growth kinetics

Efficacy studies possible within 3 weeks in OncoRat SRG

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VCaP responsive to Enzalutamide in OncoRat SRG

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12 OncoRat SRG, 10 million cells each

Treatment groups:

Group 1 : Vehicle control 15%

DMSO in PEG400 (n = 5)

Group 2 : Enzalutamide 30mg/kg

QD (n = 5)

• 83% of recipient rats enrolled in efficacy study

0 5 10 15 20 25

0

5000

10000

D a y s

TV V e h ic l e (n = 5 )

E n z a lu ta m id e (n = 5 )

OncoRat SRG for faster growth and larger tumors

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Patient Derived Xenografts [PDX]

Case study: Non-Small Cell Lung Cancer [NSCLC]

OncoRat SRG for NSCLC PDX establishment

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Growth curve for primary implants from biopsy tissue of NSCLC

OncoRat SRG for NSCLC PDX establishment

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1°Sample Recipients Take rate Cryo’d

3010 3 SRG 100% 2153067 3 SRG 67% 1203075 3 SRG 33% 1033095 3 SRG 33% 69

PDX study timeline & passage comparison

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Higher engraftment success

Resection to PDX establishment in 2 passages (3-4 months)Frontiers in Oncology. 2017. v7. article 2.

Humanizing the Immune System

ImmunoRatTM SRG

Humanization of the SRG rat immune system with PBMCs

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Immune humanization of the SRG rat with human PBMCs. 50e6 viable human PBMCs injected.Peripheral blood analyzed via FACS. By 4 weeks post-transplant, recipients had an average of 29%circulating human CD45+ cells, 46% human CD45+ cells and healthy by 10 weeks. Animals survive up to 15weeks post transplantation.

Humanization of the SRG rat immune system with PBMCs

Hera Facility & Operations

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Experienced leadership team & highly trained and certified staff

Barrier vivarium with dual HEPA filtered Innovive IVC caging system

Digital colony management & data acquisition software

Successful audit by multiple big pharma & biotech

Xenograft/PDX Efficacy Study Services

Efficacy studies

PDX establishment

Pilot tumor growth kinetics studies

OncoRat SRG rats off-the-shelf

OncoRat SRG rats pre-implanted with tumor models

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Questions?

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Feature Mouse OncoRat SRG

Engraftment Variable High

Growth profile Slow, variable Fast, uniform

Tumor size 1,000mm3 10,000+ mm3

Translational profile Not ideal More human-like

OncoRat SRG - Outperforming the mouse for xenografts

Tseten Yeshi, Ph.D.VP R&Dservices@herabiolabs.com859-414-0648