PATIENT-DERIVED TUMOR XENOGRAFTS IN HUMANIZED NSG-SGM3
MICE: A NEW IMMUNO-ONCOLOGY PLATFORM
Rick Huntress
Director of Business Development, In Vivo Pharmacology and Clinical Lab Services
PATIENT-DERIVED TUMOR XENOGRAFTS IN HUMANIZED NSGTM
MICE: A MODEL TO STUDY IMMUNE RESPONSES IN CANCER
THERAPY
James Keck, Ph.D.
Senior Director, In Vivo Pharmacology and Clinical Lab Services
Coordination Across Multiple
Campuses
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• In Vivo Efficacy
Services
• Platform Production
• PDX Resources
• JAX Clinical Genomics
• Medical Informatics
• CLIA-certified
• Targeted exome
sequencing
• Model Refinement
• Genome Informatics
Hu-NSGTM & PDX Capabilities
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• Hu-NSGTM Portfolio
– CD34+ and custom stem cells
– NSG, NSG-SGM3 and other cytokine expressing NSG derivatives
• PDX Experience
– Over 400 PDX tumors all P5 or earlier
– PDX Live
• Access options
– Delivery of models – humanized mice with or without tumors
• CD34+ are both “off the shelf” ready
– Execution of studies
• >20 IO studies
NSGTM Mice: The perfect host for
an immuno-oncology platform
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• No Mature B Cells
• No Mature T Cells
• No Natural Killer Cells
• No Complement
• Defective Dendritic Cells
• Defective Macrophages
• Low Incidence of
Lymphoma
• Median Survival >89
Weeks
>400 clinically relevant PDX tumors, with
orthotopic engraftment capabilities
PDX Models Established
Creating Humanized Mice: Timeline
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Whole body
irradiation
Tail vein
injection
Human B
cells appear
Human T
cells appear
24~48 hours 8 weeks 12 weeks3 weeks 12 weeks 15 weeks
Strain: NSG - NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (#005557)
- Highly immunodeficient
Strain: NSG-SGM3 NSG-Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ (#013062)
- Highly immunodeficient
- Improves normal human myeloid cell development after
HSC transplantation and promotes AML xenograft
efficiency.
NSG vs. NSG-SGM3
Reconstituting the human immune
system in NSG vs. NSG-SGM3 mice
Experimental Design:
Mice:
• Female
• NSG: 3-week old, n=20
• NSG-SGM3: 4-week old, n=9
Irradiation:
• NSG: 140 cGy
• NSG-SGM3: 100 cGy
HuCD34+ HSC: ~130,000 cells/mouse from the same donor
Blood collection: 4, 6, 9, 12, 15 & 18 weeks post engraftment to
check major human leukocyte lineages (flow panels: hCD45,
hCD33, hCD19, hCD3, hCD4, hCD8)
Human Immune Cell Reconstitution in the
Blood of Hu-NSG vs. Hu-NSG-SGM3
estimated
Human Immune Cell Reconstitution in the
Blood of Hu-NSG vs. Hu-NSG-SGM3
T cell populations
T cell and Myeloid
CD4 populations
Splenic immune cell populations in OMP-LU121 tumor
bearing hu-SGM3 mice 20 weeks post-CD34 engraftment
Data provided by:
Timothy Hoey and Christopher Murriel,
OncoMed Pharmaceuticals, Inc.
Employing Humanized NSG mice for Testing of
Immune Modulators That Target Human Tumors
• Engraftment with purified human CD34+ HSCs
• Confirmation of circulating human CD45+ cell population
• Human PDX tumors or human cell lines expressing PD-L1 are then
engrafted subcutaneously into the mice
• When tumors reach 70-120 mm3 mice are grouped and treated with
therapeutics including Pembrolizumab for 21 to 28 days
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Efficacy Results of Pembrolizumab +/- Docetaxel
on LG1306 PDX tumors in Hu-NSG Mice
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• Fresh tumor tissue engraftment
• HuCD45+ more than 20%
• LG1306 PD-L1 surface expression: 89.1%
HLA match
Tumor 1 2
LG1306 HLA-DRB4, DQA1, DQB1 No match
CD34+HPC donor
Significant variability in response to anti-PD1 therapeutic within a
treatment arm: LG1306 Lung PDX tumor model in Hu-NSG
Immune cell infiltration of humanized PDX
NSG™ mice in response to pembrolizumab
CD45 CD8 Cytokeratin
Vehicle Pembrolizumab
Pembrolizumab-treated mice displayed increased total immune cell and
CD8+ T lymphocyte tumor infiltration compared to vehicle-treated mice.
Preliminary Efficacy Results of LG1306 PDX in
HuCD34 NSG-SGM3 Mice
• Fresh tumor tissue
engraftment
• HuCD45+ in whole blood: 36-
81%
• HuCD3+/HuCD45: average
14.3%
• LG1306 PD-L1 surface
expression: 89.1%
LG1306 in Hu-SGM3 mice: Individual
Tumor Volume Changes
Preliminary Efficacy Results of BR1126 PDX in
HuCD34 NSG-SGM3 Mice
• Fresh tumor tissue
engraftment
• HuCD45+ in whole blood: 50-
88%
• HuCD3+/HuCD45: average
34%
• BR1126 PD-L1 surface
expression: 56.9%
BR1126 in Hu-SGM3 mice:
Individual Tumor Volume Changes
BR1126 in Hu-SGM3 mice:
Flow Data (T Cells)
BR1126 in Hu-SGM3 mice: Flow Data (PD-1)
• Engrafted with 5x106 cells/mouse s.c. with matrigel
• MDA-MB-231 cell surface expression of PD-L1: 95.1%
Evaluating the Effects of Pembrolizumab on
MDA-MB-231 Tumor Model in Hu-CD34 NSG-SGM3
MDA-MB-231 in Hu-SGM3 mice: Individual
Tumor Volume Changes
MDA-MB-231 in Hu-SGM3 mice:
Flow Data (T Cells)
MDA-MB-231 in Hu-SGM3 mice:
Flow Data (PD-1)
Preliminary Findings and
Conclusions
• PDX growth is not affected by HLA-type matching
• PDX engraftment into hu-CD34 NSGTM or hu-CD34 SGM3 mice does
not significantly impact growth kinetics
• Hu-CD34 NSGTM and hu-CD34 NSG-SGM3 mice demonstrate
immune-cell infiltration of tumors
• Hu-CD34 NSGTM and hu-CD34 NSG-SGM3 PDX respond to anti-tumor
agent Pembrolizumab
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Acknowledgements
• JAX – In Vivo Pharmacology Services
– Minan Wang, Li-Chin Yao, Mingshan Cheng and Danying Cai
• JAX – Genomic Medicine
– Karolina Palucka
• JAX – Mammalian Genetics
– Lenny Shultz, Jim Keck, Carol Bult, Susie Airhart and Ed Liu
• UMASS
– Dale Greiner and Mike Brehm
• OncoMed Pharmaceuticals, Inc.
– Timothy Hoey and Christopher Murriel,
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