A new Rag2/Il2rg SCID rat Enabling cell line xenografts ... · Rag2/Il2rg double knockout. The most...
Transcript of 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
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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
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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
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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
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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
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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
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Cell line xenografts
VCaP
OV81.2HCT116
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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
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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 )
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OncoRat SRG for faster growth and larger tumors
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Patient Derived Xenografts [PDX]
Case study: Non-Small Cell Lung Cancer [NSCLC]
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OncoRat SRG for NSCLC PDX establishment
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Growth curve for primary implants from biopsy tissue of NSCLC
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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
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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.
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Humanizing the Immune System
ImmunoRatTM SRG
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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
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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
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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]