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Predictive Cancer Models
Using Patient-Derived
Xenograft Mice
Technical Information Services
April 9, 2020
THE JACKSON LABORATORY™
To discover precise genomic
solutions for disease and
empower the global biomedical
community in our shared quest
to improve human health
Our Mission
WHAT WE DO AND PROVIDE• Biomedical Research
Genetics, biology, and mechanism of human diseases
• Mouse Models and Related Contract Research Services
Preclinical/Translational: Study-ready mouse models, mouse model generation, breeding, cryopreservation, in vivo preclinical studies– including target identification and confirmation, PK studies, efficacy studies, and more
Clinical: Specialty clinical services and assays; the JAX™ Clinical Knowledgebase (JAX-CKB)
• Education & Training
Courses, conferences, internships, pre-and post-doctoral training, live and on-
demand webinars, and on-site seminars
JAX™ MICE
• Strains developed from NIH-funding.
• >11,000 publically available mouse lines
…and growing.
>3 million mice shipped annually worldwide
• Unsurpassed genetic quality and animal
health.
• Comprehensive phenotypic descriptions
and references.
• Home to common inbred strains (C57BL/6J,
BALB/cJ, etc.) that support the
development/collection of specialty strains
and the JAX biomedical research mission.
The Gold Standard for Biomedical Research
Online Resources to Support and Expedite Your Research
JAX™ Mice Database
jax.org/mouse-search
Mouse Genome Informatics
informatics.jax.org/
Mouse Phenome Database
phenome.jax.org/
Mouse Tumor Biology Database
jax.org/mtb
JAX™ Clinical Knowledgebase
ckbhome.jax.org
Learning Goals
6
Describe how NSG™ are a
useful platform for human
disease research
Demonstrate how PDX models
are used for predictive and
preclinical research
Review Onco-Hu™ models for
immuno-oncology research
Are you currently using patient-
derived xenograft (PDX) models in
your research?
Yes
No, but I am planning to within the next 6 months
No, but I want to learn about PDX models
I do not know
7
The Need for New Preclinical Models
8
Kola 2004. Nat Rev Drug Discov. PMID:15286737
Sharpless 2006 Nat Rev Drug Discov PMID:16915232
Only 5% of cancer treatments
entering clinical trials are
approved
Major causes of attrition are
lack of efficacy (30%) and
safety (30%)
Use of more predictive animal
models is needed during
preclinical testing to diminish
attrition
Vehicle Merodantoin
Classic Xenograft Model: Nude Mouse
9
Gulliya 1994 Cancer PMID:8082074
MCF-7 breast cancer cell line
Immune System Components
10
Dendritic
cells
NSG™, a Highly Immunodeficient Mouse
NOD backgroundo Absence of hemolytic complement
o Reduced dendritic and NK cell function
o Defective macrophages
o A more human-like Sirpa allele on macrophages
11
Nomenclature
NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (005557)
JAX™ Mice |
NSG™, a Highly Immunodeficient Mouse
NOD backgroundo Absence of hemolytic complement
o Reduced dendritic and NK cell function
o Defective macrophages
o A more human-like Sirpa allele on macrophages
scid mutationo prevents development of mature T and B cells
12
Nomenclature
NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (005557)
JAX™ Mice |
NSG™, a Highly Immunodeficient Mouse
NOD backgroundo Absence of hemolytic complement
o Reduced dendritic and NK cell function
o Defective macrophages
o A more human-like Sirpa allele on macrophages
scid mutationo prevents development of mature T and B cells
gamma chain (Il2rg knockout)o Eliminates signaling from 6 distinct interleukins and blocks NK cell
development
13
Nomenclature
NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (005557)
JAX™ Mice |
Immune System Components
14
(NSGTM)
Dendritic cells
Applications and Benefitso No scid-associated leakiness
o Longer life span than NOD-scid (> 16 months)
o Enhanced primary human tumor and hematopoietic stem cell
engraftment (Sirpa allele) – immuno-oncology applications
15
Using NSG™ as a Platform for Human
Disease Research
Applications and Benefitso No scid-associated leakiness
o Longer life span than NOD-scid (> 16 months)
o Enhanced primary human tumor and hematopoietic stem cell
engraftment (Sirpa allele) – immuno-oncology applications
Considerationso Extremely immunodeficient: require aseptic housing and
handling
o scid side effects: radiation sensitivity; genotoxic drugs may
have higher toxicity
16
Using NSG™ as a Platform for Human
Disease Research
Which mouse strain is more likely
to engraft primary human cells?
BALB/c scid
NOD scid
17
JAX Patient-Derived Xenograft (PDX)
Program
18
Clinical information
o Tumor type, grade and markers (if known)
o Treatment history
Histology
Gene expression array and CNV array analysis
o Target cancer panel for mutation analysis
Archived
PDX
tumors
In Vivo Pharmacology Services |
PDX Development Process
19In Vivo Pharmacology Services |
Archived
PDX
tumors
Cohort of miceDonor mice
Multi-step approach for the development of cohorts of patient derived
xenograft mouse models
Study
Established PDX Models
20
~400 clinically relevant PDX tumors
In Vivo Pharmacology Services |
NSG™
C.B-17
scid
H&E Human cells
(anti-HLA)
Mouse blood cells
(anti-mCd45)
H&E
Cancer
Patient’s Lung
Tumor
Fragment
NSG™ Mice Preserve Patient Tumor
Characteristics
21
Simpson-Abelson 2008 J Immunol PMID:18453623
Pa
tie
nt tu
mo
rE
ng
raft
ed
tum
or
H&E Cytokeratin Ki67
NSG™ Mice Preserve Patient Tumor
Characteristics
22
Simpson-Abelson 2008 J Immunol PMID:18453623
Colon PDX Structure Fidelity
23In Vivo Pharmacology Services |
TM00388-P0TM00388-PT
Colon Adenocarcinoma Metastatic to Lung
TM00134-PT TM00134-P0 TM00134-P1
Colon Adenocarcinoma, KRAS G13D
BL0293f563P2 PDX Tumor Volume Response
to Cisplatin Single Agent Treatment (1500 mm3 tumor endpoint)
Days(Day 1 = treatment initiation)
me
an
tu
mo
r v
olu
me
(m
m3;+
/- S
E)
0 5 10 15 20 25 30 35 40 45 50 55 60 650
250
500
750
1000
1250
1500
Group 1: Saline Control
Group 2: Cisplatin 2.5 mg/kg
Bladder PDX ModelsStandard of Care Treatment Response
24In Vivo Pharmacology Services |
TM00016-P2 PDX tumor volume response to Cisplatin Single Agent
Treatment (1500 mm3 endpoint)
0 5 10 15 20 25 30 35 40 45 50 55 60 650
500
1000
1500
2000
2500
Days(Day 1 = treatment initiation)
Ind
ivid
ua
l T
um
or
Vo
lum
e (
mm
3)
Bladder PDX ModelsTumor Heterogeneity
25In Vivo Pharmacology Services |
TM00016-P2 PDX tumor volume response to Cisplatin
BL0269f404P2 PDX Tumor Volume Response
to Cisplatin Single Agent Treatment (2000 mm3 tumor endpoint)
0 10 20 30 40 500
500
1000
1500
2000
2500
Group 1: Saline Control
Group 2: Cisplatin 2.5 mg/kg (Days 1, 2, 3, 15, 16, & 17)
Days(Day 1 = treatment initiation)
me
an
tu
mo
r v
olu
me
(m
m3;+
/- S
E)
Bladder PDX Models Differential Treatment Responses
26
Model: TM00015 High grade urothelial carcinoma in bladder
(invasive)
In Vivo Pharmacology Services |
• Tumor growth rates vary
• Response varies
TM00015-P2 PDX Tumor Response to Cisplatin Treatment
PDX ModelsPlatform to test heterogeneity of tumor response
27In Vivo Pharmacology Services |
Breast Cancer PDXNSGTM support growth of ER+ breast tumors
28In Vivo Pharmacology Services |
TM00386-PT
• NSG relatively resistant to estradiol crystal formation in bladder
• Tumors retain “organoid” growth pattern
TM00386 (ER+/PR+/Her2-)
TM00386-P0 TM00386-P1
PR+ >50% ER+ 100%
Breast Cancer PDXER & PR Expression is retained
29In Vivo Pharmacology Services |
ER+/PR+/Her2-
(Estradiol supplementation)
TM00386-P1 TM00386-P1
NSCLC AdenocarcinomaDrug Susceptibility Based on Gene Expression
30
Gene
Expression
o KRAS
Gly12Cys
o EGFR wild-
type
o ERCC1low
Drug Prediction
o Resistant to
Erlotinib
o Sensitive to
Cisplatin
In Vivo Pharmacology Services |
TM00186-PT TM00186-P0
TM00186-P1 TM00186-P2
NSCLC AdenocarcinomaGene Expression Predicts Drug Susceptibility
31In Vivo Pharmacology Services |
Gene
Expression
o KRAS
Gly12Cys
o EGFR wild-
type
o ERCC1low
Drug Prediction
o Resistant to
Erlotinib
o Sensitive to
Cisplatin
Lung Adenocarcinoma EGFR L858R Acquired TKI Resistance (TM00199)
32In Vivo Pharmacology Services |
Lung Adenocarcinoma EGFR L858RPDX Tumor Models Patient Response – TM00199
33In Vivo Pharmacology Services |
34In Vivo Pharmacology Services |
huCD34+ HSCNSG™
PDX Tumor or Cell Line
Irradiation
Humanized NSG™
Humanized NSG™ with Tumor (When tumors reach 70-120 mm3 mice are treated
with therapeutics for 21 to 28 days)
Onco-Hu™
Humanized Tumor-Bearing NSG™ and NSG-SGM3™ Mice
NSG (005557)NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ
NSG-SGM3 (013062)NOD.Cg-Prkdcscid Il2rgtm1Wjl
Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ
39In Vivo Pharmacology Services |
Growth Kinetics of PDX Tumors in Hu-NSG™
PDX engrafted 12 weeks post CD34+ humanization, HuCD45+ greater than 20%
HLA mismatches (partial or full) may be tolerated
PDX growth kinetics should be tested for each
model before enrolling in a study
Due to variability in coengraftment and
therapeutic response, several CD34+ donors per experiment should be considered
Wang et al., 2018 FASEB PMID: 29146734
BR0744 Breast
LG0977 Lung
SA0209 Soft Tissue Carcinoma
Comparison of growth rate of PDX Tumors in NSG™ vs. Hu-NSG™
Keytruda and Cisplatin Inhibit Growth of
the TM00098 PDX Model in Hu-CD34
NSG™ PDX Mice
36
• Fresh tumor tissue engraftment• Hu-CD45+ in Hu-NSG™ mice:
>25% • BR1126 PD-L1 surface
expression: 56.9%
In Vivo Pharmacology Services |
Which model(s) could be useful for
cancer immunotherapy?
C57BL/6J mouse + syngeneic cancer cell line (B16-
F10)
Nude mouse + PDX + human hematopoietic stem cells
NSG mouse + PDX
NSG mouse + PDX + human hematopoietic stem cells
37
MGI: The Ultimate Laboratory
Mouse Resource
38
www.informatics.jax.org
In Vivo Pharmacology Services |
Mouse Tumor DatabasePDX Model Resource
39
www.tumor.informatics.jax.org/mtbwi/index.do
In Vivo Pharmacology Services |
Search by:
o Tumor ID
o Primary tumor site
o Diagnosis
o Gene variant
40In Vivo Pharmacology Services |
MTB: PDX Model Resource
41In Vivo Pharmacology Services |
MTB: PDX Model ResourcePDX Model Details model ID=TM00096
PDX Gene Expression Profile
42In Vivo Pharmacology Services |
PDX Model Details model ID=TM00096
Tumor Growth Characteristics
43In Vivo Pharmacology Services |
PDX Model Details model ID=TM00096
44In Vivo Pharmacology Services |
CKB molecular profile pages
MTB: Variant Summary and Clinical
Knowledgebase (CKB)
PDX Model Details model ID=TM00096
CKB: Molecular Profile
45
PDX Program Summary
46
PDX models more closely represent clinical
observations than previous models
o Gene expression, tumor structure, and treatment response
o Low-passage samples retain heterogeneity and fidelity of
human cancers
Data is available to guide model selection
Ability to pair with humanized mice for immuno-
oncology applications
In Vivo Pharmacology Services |
47
Humanized Mice, Patient-Derived Xenograft
Preclinical Models and Therapeutic Drug
Evaluation
CRISPR/Cas9 Mouse Model Generation
Common Inbred and Specialty JAX™ Mice
Aged C57BL/6J Mice (25-78 wks)
Inbred, Outbred and B6J Nude Mice
Mouse Genome Scanning
NSG™ & NRG Mouse Model Variants
Syngeneic Mice Studies for Cancer
Research (i.e., Allograft Mouse Tumor
Studies)
Mouse Cryopreservation and Recovery
Basic and Complex Mouse Breeding,
Speed Congenics, and Rederivation
Neurobiology Models and Resources
JAX™ Mice & Services: Leading Experts in Mouse Modeling
Upcoming JAX™ Webinars
Rapid in Vivo Evaluation of Immunomodulatory Therapies Using Hu-
PBMC NSG™ Mice
o April 16, 2020, 4:00 PM Central European Time
Onco-Hu™ Models: Humanized Mice for Evaluation of Immuno-
Oncology Therapeutics
o April 30, 2020, 1:00 PM USA Eastern Standard Time
www.jax.org/education-and-learning/webinars |
Subscribe to the monthly webinar announcements email list:
https://subscribe.jax.org/
49
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questions to [email protected]
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JAX™ Mice, Clinical & Research Services
1-800-422-6423 (USA) • 1-207-288-5845
(international)
www.jax.org/jax-mice-and-services
