utilizing humanized nsg mice to evaluate drug efficacy...
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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
NSG™ vs. NSG™-SGM3 Mice
NOD scid gamma (NSG™)
NOD.Cg-Prkdcscid Il2rgtm1Wjl/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-Prkdcscid Il2rgtm1Wjl 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. 2010. Leukemia 24(10):1785-8. PMID: 20686503
Billerbeck E. et al. 2011. Blood 117(11):3076-86. PMID: 21252091
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Experimental Timeline for Humanization
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Human cord blood derived CD34+ cells engrafted in female mice
Mature T cell
development
Whole body irradiation
Tail vein injection
3 weeks
4 weeks 12 weeks ~18 weeks
~18 weeks
NSG
NSG-SGM3
In Vivo Pharmacology Services |
Validation of
Engraftment by
FACS
16 weeks
B cells mature ahead of T cells
Myeloid cells peak at 15 weeks in
NSG-SGM3
Human Immune Cells in Peripheral Blood of Hu-
NSG™ vs. Hu-NSG™-SGM3: Absolute Counts
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Total Human Donor (cells/µl)
HuCD33 Myeloid Cells (cells/µl)
In Vivo Pharmacology Services |
Human Immune Cells in Peripheral Blood of Hu-
NSG™ vs. Hu-NSG™-SGM3: Absolute Counts
6
HuCD19 B Cells (cells/µl)
HuCD3 T Cells (cells/µl)
In Vivo Pharmacology Services |
Human Immune Cells in Peripheral Blood of Hu-
NSG™ vs. Hu-NSG™-SGM3: Absolute Counts
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HuCD8 Cytotoxic T Cells (cells/µl)
HuCD4 Helper T Cells (cells/µl)
In Vivo Pharmacology Services |
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
Humanization of NSG™ Mice Has No
Significant Impact on PDX Growth Kinetics
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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%
PDX Tumors Are Infiltrated With
Human Immune Cells
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HuCD45+ in BR0744 Tumor
Hu-N
SG
NSG
0.0
0.5
1.0
1.5
2.0
% o
f h
CD
45+
cells
No passenger leukocytes in PDX tumors
Tumors infiltrated with engrafted human cells
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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
Hu-NSG™ MDA-MB-231 Mice: Efficacy
Requires the engrafted immune system
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MDA-MB-231 Tumor Response in Non-Humanized Mice
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DAPI CD8
Pembrolizumab Vehicle
Hu-NSG™ MDA-MB-231 Mice: Pembrolizumab Initiates CD8+ T Cell Infiltration
Hu-NSG™ MDA-MB-231 Mice: Efficacy Of
Pembrolizumab Is CD8+ T Cell Dependent
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MDA-MB-231 Tumor Response inHu-NSG Mice
0 10 20 300
200
400
600
800Vehicle (Q5DX6)+isotype (Q7DX5)
Pembrolizumab (Q5Dx6)+isotype (Q7DX5)
Pembrolizumab (Q5DX6)+anti-CD8 (Q7DX5)
Days
Tu
mo
r vo
lum
e(m
m3)+
SE
M
Hu-NSG™ MDA-MB-231 Mice: Characterization
of Human Tumor Infiltrating Cells & PD-1 Levels
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% PD-1 on HuCD45 Cells in Tumor
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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 |
24 In Vivo Pharmacology Services |
Hu-NSG™ BR1126 TNBC PDX Mice:
Pembrolizumab Inhibits Tumor Growth
HLA match
Tumor 1 2 3
BR1126 HLA-C, DPA1 HLA-A,DQA1, DPB1, DPA1 HLA-C, DPA1
CD34+HPC donor
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BR1126 in Hu-NSGTM Mice: Flow Data (T Cells) No Correlation of Human Immune Cell
Infiltration with Tumor Growth Rate Reduction
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Hu-NSG™-SGM3 BR1126 TNBC PDX Mice:
Pembrolizumab & Doxorubicin Inhibit
Tumor Growth
In Vivo Pharmacology Services |
Evaluating the effects of PD-1 inhibitors on
MDA-MB-231 tumor model in Hu-NSGTM
• Nivolumab and Pembrolizumab – matched efficacy
30 In Vivo Pharmacology Services |
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%
Hu-NSG™ MDA-MB-231 Mice: Suppression
of Breast Tumor Growth by Anti-OX40
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0 5 1 0 1 5 2 0 2 5 3 0 3 5
0
2 0 0
4 0 0
6 0 0
8 0 0
1 0 0 0
T w o D o n o r s ( # 5 0 3 8 , # 5 0 4 0 )
S t u d y D a y s
Me
an
T
um
or
Vo
lum
e (
mm
3)
+/-
SE
M
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 < 0 . 0 5
* * p < 0 . 0 1
* * * p < 0 . 0 0 5
* * * * p < 0 . 0 0 0 1
2 - t a i l e d u n p a i r e d t - t e s t
* *
*
* * *
*
Jean Gudas VP, Research & Development ImaginAb, Inc.
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*p≤0.009 21MR ± PD1 vs Control, D11 **p≤0.0006 21MR ± PD1 vs Control, D19
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
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
HLA match
Tumor CD34+ HPC Donor
LG1208 HLA-DRB4, DPA1
Hu-NSG™ LG1208 NSCLC Lung PDX Mice: No Inhibition of Tumor Growth By Pembrolizumab
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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
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
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 anti-
tumor agents; anti-PD-1, anti-CTLA4, anti-OX40, and anti-DLL4
37 In Vivo Pharmacology Services |
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
In need of humanized
mice to advance your
research?
39
Contact JAX Technical Support
https://www.jax.org/jax-mice-and-services/customer-
support/technical-support
JAX® Mice, Clinical & Research Services
1-800-422-6423 • 1-207-288-5845
[email protected] • www.jax.org/jaxmice
12-16 wks