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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

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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

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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

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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

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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

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>400 clinically relevant PDX tumors, with

orthotopic engraftment capabilities

PDX Models Established

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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

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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

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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)

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Human Immune Cell Reconstitution in the

Blood of Hu-NSG vs. Hu-NSG-SGM3

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estimated

Human Immune Cell Reconstitution in the

Blood of Hu-NSG vs. Hu-NSG-SGM3

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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.

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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

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Significant variability in response to anti-PD1 therapeutic within a

treatment arm: LG1306 Lung PDX tumor model in Hu-NSG

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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.

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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%

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LG1306 in Hu-SGM3 mice: Individual

Tumor Volume Changes

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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%

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BR1126 in Hu-SGM3 mice:

Individual Tumor Volume Changes

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BR1126 in Hu-SGM3 mice:

Flow Data (T Cells)

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BR1126 in Hu-SGM3 mice: Flow Data (PD-1)

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• 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

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MDA-MB-231 in Hu-SGM3 mice: Individual

Tumor Volume Changes

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MDA-MB-231 in Hu-SGM3 mice:

Flow Data (T Cells)

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MDA-MB-231 in Hu-SGM3 mice:

Flow Data (PD-1)

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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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