Utilizing Humanized NSG Mice to Evaluate Drug Efficacy in Immuno-Oncology
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1 Utilizing Humanized NSG Mice to Evaluate Drug Efficacy in Immuno-Oncology Rick Huntress Director Business Development March 15, 2017
2 Onco-Hu : Humanized Mice for Evaluation of Immuno-Oncology Therapeutics Characteristics of humanized NSG and NSG -SGM3 mice PDX Tumor Growth Mechanism of Action Efficacy data CD34+ Donor Variability In Vivo Pharmacology Services 2
3 NSG vs. NSG -SGM3 Mice NOD scid gamma (NSG ) NOD.Cg-Prkdc scid Il2rg tm1wjl /SzJ (005557) The current gold standard for reconstitution of the human immune system Excellent functional T cell responses Limited human myeloid lineage development NSG -SGM3 NOD.Cg-Prkdc scid Il2rg tm1wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ (013062) Promotes improved AML engraftment efficiency Improves normal human myeloid cell development after HSC transplantation Wunderlich M. et al Leukemia 24(10): PMID: Billerbeck E. et al Blood 117(11): PMID:
4 Experimental Timeline for Humanization B cells mature ahead of T cells Myeloid cells peak at 15 weeks in NSG-SGM3 Whole body irradiation Tail vein injection Validation of Engraftment by FACS Mature T cell development NSG NSG-SGM3 3 weeks 16 weeks 4 weeks 12 weeks ~18 weeks ~18 weeks Human cord blood derived CD34 + cells engrafted in female mice In Vivo Pharmacology Services 4
5 Human Immune Cells in Peripheral Blood of Hu- NSG vs. Hu-NSG -SGM3: Absolute Counts Total Human Donor (cells/µl) HuCD33 Myeloid Cells (cells/µl) In Vivo Pharmacology Services 5
6 Human Immune Cells in Peripheral Blood of Hu- NSG vs. Hu-NSG -SGM3: Absolute Counts HuCD19 B Cells (cells/µl) HuCD3 T Cells (cells/µl) In Vivo Pharmacology Services 6
7 Human Immune Cells in Peripheral Blood of Hu- NSG vs. Hu-NSG -SGM3: Absolute Counts HuCD4 Helper T Cells (cells/µl) HuCD8 Cytotoxic T Cells (cells/µl) In Vivo Pharmacology Services 7
8 Onco-Hu : Humanized Mice for Evaluation of Immuno-Oncology Therapeutics Characteristics of humanized NSG and NSG -SGM3 mice PDX Tumor Growth Mechanism of Action Efficacy data CD34+ Donor variability In Vivo Pharmacology Services 8
9 Humanization of NSG Mice Has No Significant Impact on PDX Growth Kinetics No difference between NSG and Hu-NSG mice on tumor growth curve No HLA match testing performed Fresh tumor tissue engraftment 100% take rate in NSG or Hu-NSG mice HuCD45+ more than 20% 9
10 % of hcd45+ cells PDX Tumors Are Infiltrated With Human Immune Cells 2.0 HuCD45 + in BR0744 Tumor Hu-NSG NSG No passenger leukocytes in PDX tumors Tumors infiltrated with engrafted human cells 10
11 MDA-MB-231 Tumor Grows Slower in Hu-NSG -SGM3 Than in Hu-NSG 11
12 Onco-Hu : Humanized Mice for Evaluation of Immuno-Oncology Therapeutics Characteristics of humanized NSG and NSG -SGM3 mice PDX Tumor Growth Mechanism of Action Efficacy data CD34+ Donor Variability In Vivo Pharmacology Services 12
13 Hu-NSG MDA-MB-231 Mice: Efficacy Requires the engrafted immune system MDA-MB-231 Tumor Response in Non-Humanized Mice 13
14 Hu-NSG MDA-MB-231 Mice: Suppression of Breast Tumor Growth by Pembrolizumab
15 MDA-MB-231 in Hu-NSG TM Mice: Flow Data (T Cells) Tumor Infiltration Levels are Variable
16 Hu-NSG MDA-MB-231 Mice: Pembrolizumab Initiates CD8 + T Cell Infiltration Vehicle Pembrolizumab DAPI CD8 16
17 Tumor volume(mm 3 )+SEM Hu-NSG MDA-MB-231 Mice: Efficacy Of Pembrolizumab Is CD8 + T Cell Dependent MDA-MB-231 Tumor Response in Hu-NSG Mice Vehicle (Q5DX6)+isotype (Q7DX5) Pembrolizumab (Q5Dx6)+isotype (Q7DX5) Pembrolizumab (Q5DX6)+anti-CD8 (Q7DX5) Days 17
18 Hu-NSG MDA-MB-231 Mice: Characterization of Human Tumor Infiltrating Cells & PD-1 Levels % PD-1 on HuCD45 Cells in Tumor 18
19 Onco-Hu : Humanized Mice for Evaluation of Immuno-Oncology Therapeutics Efficacy with a checkpoint inhibitor requires an engrafted immune system CD8+ T cell infiltration is increased with anti PD-1 CD8 Depletion blocks checkpoint efficacy Drug is binding the intended target (PD-1) In Vivo Pharmacology Services 19
20 Onco-Hu : Humanized Mice for Evaluation of Immuno-Oncology Therapeutics Characteristics of humanized NSG and NSG -SGM3 mice PDX Tumor Growth Mechanism of Action Efficacy data CD34+ Donor Variability In Vivo Pharmacology Services 20
21 Hu-NSG -SGM3 MDA-MB-231 Mice: Suppression of Breast Tumor Growth by Pembrolizumab In Vivo Pharmacology Services 21
22 MDA-MB-231 in Hu-SGM3 Mice: Flow Data (T Cells)
23 MDA-MB-231 in Hu-SGM3 Mice: Flow Data (PD-1)
24 Hu-NSG BR1126 TNBC PDX Mice: Pembrolizumab Inhibits Tumor Growth HLA match CD34 + HPC donor Tumor BR1126 HLA-C, DPA1 HLA-A,DQA1, DPB1, DPA1 HLA-C, DPA1 In Vivo Pharmacology Services 24
25 BR1126 in Hu-NSG TM Mice: Flow Data (T Cells) No Correlation of Human Immune Cell Infiltration with Tumor Growth Rate Reduction 25
26 Hu-NSG -SGM3 BR1126 TNBC PDX Mice: Pembrolizumab & Doxorubicin Inhibit Tumor Growth In Vivo Pharmacology Services 26
27 BR1126 in Hu-SGM3 Mice: Flow Data (T Cells) 27
28 BR1126 in Hu-SGM3 Mice: Flow Data (PD-1) 28
29 Evaluating the effects of PD-1 inhibitors on MDA-MB-231 tumor model in Hu-NSG TM Nivolumab and Pembrolizumab matched efficacy
30 Hu-NSG -SGM3 LG1306 Lung PDX Mice: Pembrolizumab & Ipilimumab Inhibit Tumor Growth HuCD45+ in whole blood: 36-81% HuCD3+/HuCD45: average 14.3% LG1306 PD-L1 surface expression: 89.1% In Vivo Pharmacology Services 30
31 M e a n T u m o r V o l u m e ( m m 3 ) + / - S E M Hu-NSG MDA-MB-231 Mice: Suppression of Breast Tumor Growth by Anti-OX40 Jean Gudas VP, Research & Development ImaginAb, Inc. T w o D o n o r s ( # , # ) * * G r o u p 1 ( V e h i c l e ) G r o u p 3 ( a n t i - O X 4 0 ) * * * * * * p < * * p < * * * p < * * * * p < * 2 - t a i l e d u n p a i r e d t - t e s t S t u d y D a y s 31
32 Hu-NSG -SGM3 OMP-LU121 NSCLC Mice: Demcizumab (21MR) Inhibits Tumor Growth Data aquired in collaboration with Chris Murriel &Tim Hoey, OncoMed Pharmaceuticals OMP-LU121 NSCLC did not respond to Pembrolizumab in Hu-NSG-SGM3 mice Demcizumab (21MR) alone suppressed progression of tumor growth *p MR ± PD1 vs Control, D11 **p MR ± PD1 vs Control, D19 32
33 Anti-DLL4 Significantly Inhibits OMP-LU121 NSCLC AdC Growth in Hu-SGM3 Mice
34 Onco-Hu : Humanized Mice for Evaluation of Immuno-Oncology Therapeutics Characteristics of humanized NSG and NSG -SGM3 mice PDX Tumor Growth Mechanism of Action Efficacy data CD34+ Donor Variability In Vivo Pharmacology Services 34
35 Hu-NSG LG1208 NSCLC Lung PDX Mice: No Inhibition of Tumor Growth By Pembrolizumab HLA match Tumor LG1208 CD34+ HPC Donor HLA-DRB4, DPA1 35
36 Hu-NSG LG1208 NSCLC Lung PDX Mice: PD-1 and PD-L1 Expression in Tumor Tissue Pembrolizumab entered tumors and bound to CD45+ PD-1+ leukocytes PD-1 Expression: Activated T cells, Tregs, B cells, NK cells and monocytes; occasionally on tumor cells PD-L1 Expression: Mainly on tumor cells and normal tissues; also on T cell, B cells, macrophages, DCs 36
37 Immuno-Oncology Summary NSG and NSG -SGM3 are a proven platform for engraftment of the human immune system PDX growth is not grossly effected by HLA-type matching o Tumor growth kinetics are similar in humanized and non-humanized hosts o ~15% of PDX tumors fail to grow in humanized mice Human immune effector cells infiltrate human tumors in both Hu-CD34-NSG and Hu-CD34-SGM3 o CD4 and CD8 T cells and CD19 B cells Hu-CD34-NSG and Hu-CD34-SGM3 PDX respond to antitumor agents; anti-pd-1, anti-ctla4, anti-ox40, and anti-dll4 In Vivo Pharmacology Services 37
38 Acknowledgements JAX In Vivo Pharmacology Services o James Keck, Minan Wang, Li-Chin Yao, Mingshan Cheng and Danying Cai JAX Genomic Medicine o Karolina Palucka JAX Mammalian Genetics o Lenny Shultz, Brian Soper, Carol Bult, Susie Airhart and Ed Liu UMASS o Dale Greiner and Mike Brehm 38
39 In need of humanized mice to advance your research? wks Contact JAX Technical Support JAX Mice, Clinical & Research Services
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