A new Rag2/Il2rg SCID rat. Enabling cell line xenografts. Accelerating PDX establishment. Humanizing the immune system
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1 A new Rag2/Il2rg SCID rat Enabling cell line xenografts Accelerating PDX establishment Humanizing the immune system Tseten Yeshi, Ph.D. VP R&D
2 About Hera BioLabs SRG Novel Technologies Proprietary Platforms Strong R&D Pipeline piggybac TM Cas-CLOVER TM The SRG Rat Engineered Cell Lines Humanized Immune System Humanized Liver
3 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 3
4 The SRG platform 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.
5 Immunophenotype of SRG rats Wild type rat SRG rat 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. 5
6 NK Cells in SRG vs. Nude Rats Nude rat SRG rat Comparison between Nude rats and the SRG rat show a severally reduced number of natural killer (NK) cells in the circulating blood. 6
7 Outperforming the mouse for xenografts Feature Mouse OncoRat SRG Engraftment Variable High Growth profile Slower, variable Faster, uniform Tumor size 1,000mm 3 10,000+ mm 3 Physiology Not ideal More human-like 7
8 Cell line xenografts VCaP OV81.2 HCT116
9 Enabling Case Study: VCaP in OncoRat SRG 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 Individual tumor volume in OncoRat SRG Serum PSA levels correlative to tumor volume 9
10 Case Study: VCaP in OncoRat SRG Highly improved engraftment efficiency and growth kinetics Efficacy studies possible within 3 weeks in OncoRat SRG 10
11 TV VCaP responsive to Enzalutamide in OncoRat SRG OncoRat SRG, 10 million cells each Treatment groups: 5000 V e h ic le (n = 5 ) E n z a lu ta m id e (n = 5 ) 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 D a y s 11
12 OncoRat SRG for faster growth and larger tumors 12
13 Patient Derived Xenografts [PDX] Case study: Non-Small Cell Lung Cancer [NSCLC]
14 OncoRat SRG for NSCLC PDX establishment Growth curve for primary implants from biopsy tissue of NSCLC 14
15 OncoRat SRG for NSCLC PDX establishment 1 Sample Recipients Take rate Cryo d SRG 100% SRG 67% SRG 33% SRG 33% 69 15
16 PDX study timeline & passage comparison Higher engraftment success Resection to PDX establishment in 2 passages (3-4 months) Frontiers in Oncology v7. article 2. 16
17 Humanizing the Immune System ImmunoRat TM SRG
18 Humanization of the SRG rat immune system with PBMCs Humanization of the SRG rat immune system with PBMCs 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 15 weeks post transplantation. 18
19 Hera Facility & Operations 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 19
20 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 20
21 Questions? OncoRat SRG - Outperforming the mouse for xenografts Feature Mouse OncoRat SRG Engraftment Variable High Growth profile Slow, variable Fast, uniform Tumor size 1,000mm 3 10,000+ mm 3 Translational profile Not ideal More human-like Tseten Yeshi, Ph.D. VP R&D services@herabiolabs.com
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