a new rag2/il2rg scid rat enabling cell line xenografts ... · rag2/il2rg double knockout. the most...
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A new Rag2/Il2rg SCID rat
Enabling cell line xenografts
Accelerating PDX establishment
Humanizing the immune system
Tseten Yeshi, Ph.D.
VP R&D
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&[email protected]