Modeling Human Cancer in Mice 40x 10x · Modeling Human Cancer in Mice Xenograft Models Xenograft...
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Modeling Human Cancer in Mice 40x 10x Presented by: Brian W. Soper, PhD Technical Information Services Hosted by Charles River, exclusive distributor of JAX ® Mice in 27 European Countries
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Transcript of Modeling Human Cancer in Mice 40x 10x · Modeling Human Cancer in Mice Xenograft Models Xenograft...
Modeling Human
Cancer in Mice 40x
10x
Presented by:
Brian W. Soper, PhD
Technical Information Services
Hosted by Charles River, exclusive
distributor of JAX® Mice in 27 European
Countries
The Jackson Laboratory’s Mission
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Performing Research
Investigating genetics and biology of human disease
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JAX® Mice The Gold Standard for Biomedical Research
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Common inbred strains (C57BL/6J, BALB/cJ, DBA/2J) support
development/collection of specialty strains and other valuable
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Online Resources to Expedite
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Online Resources to Expedite
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www.informatics.jax.org
www.jax.org/jaxmice/support/techsupport-index |
Online Resources to Expedite
Research
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www.jax.org/jaxmice
Mouse Genome Informatics
www.informatics.jax.org
Mouse Phenome Database
www.jax.org/phenome
And many more unique
resources
www.jax.org/jaxmice/support/techsupport-index |
ACS Cancer Incidence USA Statistics through 2011
7
www.cancer.org/research/cancerfactsfigures
Females Males
Rate
per
100,0
00
Lung
1970 2011
Prostate
20
40
60
80
20
40
60
80
1970 2011
Breast Lung
JAX® Mice |
Modeling Human Cancer in Mice
Xenograft Models
Xenograft Host Selection
Spontaneous and Inducible Models
Online Data and Resources
JAX® Mice | 8
Utilizing Xenograft Models
Xenograft Sample
o Established cell lines
o Primary cell lines
o Patient derived tumor tissue
Immunodeficient Hosts
o Nude
o NOD-scid
o NOD-scid gamma (NSG)
JAX® Mice | 9
Utilizing Xenograft Models
Xenograft Sample
o Established cell lines
o Primary cell lines
o Patient derived tumor tissue
Immunodeficient Hosts
o Nude
o scid
o NOD-scid gamma (NSG)
JAX® Mice | 10
JAX® Mice | 11
Immune System Components
JAX® Mice | 12
Inbred and Outbred Nude Mice
Mouse Nomenclature
Inbred Nude NU/J (002019)
Outbred Nude J:NU (007850)
Immunological Deficiencies
Mutation: Foxn1nu
Athymic and T cell Deficient
Flanagan SP. 1966. Genet Res 8(3):295-309. PMID:5980117
Pantelouris EM. 1973. Differentiation 1(6):437-50. PMID:4547146
JAX® Mice | 13
Inbred and Outbred Nude Mice
Mouse Nomenclature
Inbred Nude NU/J (002019)
Outbred Nude J:NU (007850)
Immunological Deficiencies
Mutation: Foxn1nu
Athymic and T cell Deficient
Applications and Benefits
Engraftment of human & mouse tumor cell lines
Well published/characterized
Hairless phenotype enhances assessment of tumor growth
Flanagan SP. 1966. Genet Res 8(3):295-309. PMID:5980117
Pantelouris EM. 1973. Differentiation 1(6):437-50. PMID:4547146
JAX® Mice | 14
Immune System Components
JAX® Mice | 15
Easy Evaluation of Therapeutic
Response in Nude Mice
Gulliya KS et al. 1994. Cancer 74(6):1725-32. PMID:8082074
Before treatment After treatment
JAX® Mice | 16
Nude Mice
Mouse Nomenclature
Inbred Nude NU/J (002019)
Outbred Nude J:NU (007850)
Immunological Deficiencies
Mutation: Foxn1nu
Athymic and T cell deficient
Considerations
Innate immunity intact
Not useful for hematopoietic cancer models
Not suitable for primary cell transplantation
Flanagan SP. 1966. Genet Res 8(3):295-309. PMID:5980117
Pantelouris EM. 1973. Differentiation 1(6):437-50. PMID:4547146
17 JAX® Mice |
BALB/c-scid
Mouse Nomenclature
CBySmn.CB17-Prkdcscid/J (001803)
o Similar to original C.B-17 scid
Immunological Deficiencies
Mutation: scid “severe combined immune deficiency”
No mature B and T cells
JAX® Mice | 18
BALB/c-scid
Mouse Nomenclature
CBySmn.CB17-Prkdcscid/J (001803)
o Similar to original C.B-17 scid
Immunological Deficiencies
Mutation: scid “severe combined immune deficiency”
No mature B and T cells
Applications
Xenograft host for cancer cell lines
Efficacy testing of therapeutic antibodies (CDC and ADCC intact)
Adoptive transfer from BALB/c donors
Infectious disease
Common inbred background simplifies creation of compound
immunodeficient mutants
JAX® Mice | 19
Immune System Components
JAX® Mice | 20
BALB/c-scid
Therapeutic Ab/Fusion Protein Efficacy
Shin S et al. 2011. Mol Cancer Ther 10:603-614. PMID:21393427
JAX® Mice | 21
Human SK-BR-3 Breast Cancer Cells
Mouse Nomenclature CBySmn.CB17-Prkdcscid/J (001803) o Similar to original C.B-17 scid
Applications Xenograft host for cancer cell lines
Efficacy testing of therapeutic antibodies
Adoptive transfer from BALB/c donors
Common inbred background simplifies creation of compound immunodeficient mutants
Considerations Innate immunity intact
scid side effects: radiation sensitivity; genotoxic drugs may have higher toxicity
High scid leakiness; mice develop T and B cells as they age
BALB/c-scid
JAX® Mice | 22
NOD-scid
Mouse Nomenclature NOD.CB17-Prkdcscid/J (001303)
Immunological Deficiencies Mutation: scid “severe combined immune deficiency”
No mature B and T cells
Reduced innate immunity o macrophage, dendritic, and NK cell deficits
o complement deficient
o Sirpα allele
Applications Xenotransplantation of human tumors
Engrafts some hematopoietic cancer cell lines
Adoptive transfer recipient for study of autoimmune type 1 diabetes
Significantly less scid leakiness than other backgrounds
Shultz LD et al. 1995. J Immunol 154(1):180-91.PMID:7995938
Banuelos SJ et al. 2004. Clin Immunol 112(3):273-83. PMID:15308121
JAX® Mice | 23
Immune System Components
JAX® Mice | 24
Immune System Components
JAX® Mice | 25
NOD-scid Mice Propagate
Medulloblastoma-like Tumors
Werbowetski-Ogilvie TE et al. 2012. Stem Cells 30(3):392-404 PMID:22213600
JAX® Mice | 26
Arrows point to
neural rosettes
NOD-scid
Mouse Nomenclature
NOD.CB17-Prkdcscid/J (001303)
Immunological Deficiencies
Mutation: scid “severe combined immune deficiency”
No mature B and T cells
Reduced innate immunity
Considerations
Short life span (thymic lymphoma by ~9 months
Residual innate immunity (NK cell function)
scid side effects: radiation sensitivity; genotoxic drugs may have
higher toxicity Shultz LD et al. 1995. J Immunol 154(1):180-91. PMID:7995938
Banuelos SJ et al. 2004. Clin Immunol 112(3):273-83. PMID:15308121
JAX® Mice | 27
NSG, NOD scid gamma
Mouse Nomenclature
NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (005557)
Immunological Deficiencies
NOD background contributes innate immune deficiencies
o Absence of hemolytic complement
o Reduced dendritic cell function
o Defective macrophages
o Sirpα allele
scid mutation prevents maturation of T and B cells
Il2rg deficiency eliminates signaling from 6 distinct interleukins
and blocks NK cell development
JAX® Mice | 28
NSG, NOD scid gamma Mouse Nomenclature
NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (005557)
Immunological Deficiencies
NOD background contributes innate immune deficiencies
o Absence of hemolytic complement
o Reduced dendritic cell function
o Defective macrophages
o Sirpα allele
scid mutation prevents maturation of T and B cells
Il2rg deficiency eliminates signaling from 6 distinct interleukins
and blocks NK cell development
Benefits No scid-associated leakiness Longer life span than NOD-scid (> 16 months) Enhanced human hematopoietic stem cell engraftment (Sirpa allele)
JAX® Mice | 29
Immune System Components
JAX® Mice | 30
NSG, NOD scid gamma Mouse Nomenclature
NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (005557)
Immunological Deficiencies
NOD background contributes innate immune deficiencies
scid mutation prevents development of mature T and B cells
Il2rg deficiency eliminates signaling from 6 distinct interleukins
and blocks NK cell development
Benefits
Optimal human hematopoietic stem cell engraftment (Sirpa allele)
No scid-associated leakiness
Longer life span than NOD-scid (> 16 months)
Limitations/Considerations
scid side effects: radiation sensitivity; genotoxic drugs may have
higher toxicity
JAX® Mice | 31
NSG: Improved Melanoma
Metastasis Model
Carreno et al. 2008. Clin Cancer Res 15(10):3277-86. PMID:19447870
Cell line:
o A375 human malignant melanoma
o Luciferase tagged
o Injected I.V.
JAX® Mice | 32
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
Simpson-Abelson MR et al. 2008.J Immunol 180(10):7009-18. PMID:18453623
JAX® Mice | 33
Pati
en
t tu
mo
r E
ng
raft
ed
tum
or
H&E Cytokeratin Ki67 Simpson-Abelson MR et al. 2008.J Immunol 180(10):7009-18. PMID:18453623
JAX® Mice | 34
NSG Mice Preserve Patient Tumor
Characteristics
JAX Patient-Derived Xenograft
(PDX) Program
In Vivo Pharmacology Services |
Collect primary human patient samples and engraft into NSG mice
Model Supporting Data
Clinical information:
o Tumor type, grade and markers (if known)
o treatment history
Histology
Gene expression array data
CNV array analysis
35
Access supporting data at
www.tumor.informatics.jax.org/mtbwi/pdxSearch.do
JAX Patient-Derived Xenograft
(PDX) Program
In Vivo Pharmacology Services | 36
Estrogen Receptor Positive
Breast Carcinoma
NSG supports growth of ER+ breast tumors in combination with estrogen pellets
Tumors retain “organoid” growth pattern
TM00386 PT TM00386-294 P0
PT= Patient Tumor; P0 = first patient derived xenograft (PDX) generation in mouse;
P1 = 2nd generation
TM00386-294 P1
TM00386 (ER+/PR+/Her2-)
In Vivo Pharmacology Services | 37
Spontaneous and Induced Models
Prominent and Newly Emerging Models
Breast Cancer
Lung Cancer
Prostate Cancer
Intestinal/Colon Cancer
Melanoma
Pancreatic Cancer
JAX® Mice | 38
Mammary Cancer Model FVB/N-Tg(MMTV-PyVT)634Mul/J (002374)
B6.FVB-Tg(MMTV-PyVT)634Mul/LellJ (022974)
Guy CT et al. 1992. Mol Cell Biol 12(3):954-61. PMID:1312220
Lifsted T et al. 1998. Int J Cancer 77(4):640-4. PMID:9679770
Davie SA et al. 2007 Transgenic Res 16(2):193-201. PMID: 17206489
JAX® Mice | 39
FVB/N Characteristics:
100% incidence
o Females: median age 53 days
o Pregnancy accelerates
tumorigenesis
Metastasize to the lung
o 80-94% of females
B6 Characteristics:
100% incidence
o Females: median age 92 days
Metastasize to the lung
o Reduced compared to FVB/N
background
Mammary Cancer Model STOCK Trp53tm1Brd Brca1tm1Aash Tg(LGB-cre)74Acl/J
(012620)
Characteristics:
Trp53 -/+, floxed Brca1, Lactoglobulin β-cre
64% of mice develop mammary gland tumors
o Human basal-like breast cancer
o High grade DCIS adjacent to invasive tumor
o Metaplastic element
Two rounds of breeding required
o Hormone accelerated phenotype
o Tumor development by 31 weeks
McCarthy A. TV et al. 2007. J Path 211(4):389-98. PMID:17212342
40 JAX® Mice |
High Grade Invasive
Ductal Carcinoma
Nuclear
Pleomorphism
Early-Stage Lung Cancer Model B6.129S4-Krastm4Tyj/J (008179)
Characteristics:
Floxed-stop mutant oncogenic
KrasG12D
Cre introduced nasally
(Adeno-cre)
Lesions induced 6 weeks post-cre
administration
Adenocarcinomas by 16 weeks
post-cre administration
Progressive lung disease: atypical adenomatous
hyperplasia to adenoma
Jackson EL et al. 2001. Genes Dev 15(24):3243-8. PMID: 11751630.
Meylan E et al. 2009. Nature 462(7269):104-7. PMID: 19847165.
5x105 pfu @ 6wks 5x106 pfu @ 6wks
Prostate Cancer Models TRAMP strains
Greenberg NM et al. 1995. PNAS 92(8):3439-43. PMID:7724580 Gingrich JR et al. 1996. Cancer Res 56(18):4096-102. PMID:8797572
Albertelli et al. 2006. Mol Endocrinol 20(6):1248-60. PMID:16601069
TRansgenic Adenocarcinoma Mouse Prostate
C57BL/6-Tg(TRAMP)824Ng/J (003135)
(C57BL/6-Tg(TRAMP)8247Ng/J X FVB/NJ)F1/J (008215)
Characteristics: o Background influences tumor
development
o F1 shorter lifespan
F1 ~19 wks
B6 ~52 wks
o 100% metastatic in F1
Neuroendocrine cells
Lungs or lymph nodes
About 28 weeks of age
o SV40 Tumor-antigen
o Variable tumor latency
Hyperplasia
(~10 weeks)
Adenocarcinoma
(~18wks)
JAX® Mice | 42
Inducible Prostate Cancer Models FVB;B6-Tg(Pbsn-cre)20Fwan/J (023325)
43
Publication involving:
B6.129S4-Krastm4Tyj/J (008179)
Scrib loxp-flanked mice
Probasin cre (Pbsn-cre) alone does not develop tumors
Pbsn-cre::KrasG12D/+::Scribfl/fl generates invasive carcinoma (~400 days)
Pearson HB et al. 2011. J Clin Invest 121(11):4257-67. PMID: 21965329
PIN = prostate intraepithelial neoplasia
Intestinal / Colon Cancer Model C57BL/6J-ApcMin/J (002020)
Adenomatosis Polyposis Coli, Multiple Intestinal Neoplasia
Characteristics:
Adenomas to adenocarcinomas
o Spontaneous, 100% incidence
o Age of onset: 2.6–8.7 months
o Higher polyp count on high fat diet
Females unhealthy
o Severely anemic & die by 4 months
o 12% mammary tumor incidence
Homozygous lethal e7.0 Moser AR et al. 1990. Science 247:322-4. PMID:2296722
Su LK et al. 1992. Science 256:1114. PMID:1350108
Normal intestinal tract
Multiple Adenomas
(131 days)
JAX® Mice | 44
Melanoma Model STOCK Braftm1Mmcm Ptentm1Hwu
Tg(Tyr-cre/ERT2)13Bos/BosJ (013590)
Dankort et al. 2009. Nature Genetics 41(5):544-52. PMID:19282848
Damsky et al. 2011. Cancer Cell 20(6):741-54. PMID:22172720
45 JAX® Mice |
Floxed conversion to BrafV600E
Floxed Pten KO
Inducible melanomas in 3-4 weeks
25% spontaneous melanomas in 2-12 mo.
Skin
Lung
Pancreatic Cancer Modeling B6.FVB-Tg(Pdx1-cre)6Tuv/J (014647)
Hill et al. 2010. Cancer Res 10(18):7114-24. PMID:20807812
Triple compound mutant mice develop
pancreatic ductal adenocarcinoma
Recent publication involving:
C;129S4-Ptentm1Hwu/J (004597)
B6.129S4-Krastm4Tyj/J (008179)
JAX® Mice | 46
Pdx1 promoter; pancreatic and duodenal homeobox 1
Additional Models Available
www.jax.org/jaxmice/cancer/index.html
47
Mouse Genome Informatics www.informatics.jax.org
JAX® Mice | 48
Mouse Tumor Biology (MTB) Database www.informatics.jax.org
JAX® Mice | 49
Mouse Tumor Biology (MTB) Database
50
http://tumor.informatics.jax.org
JAX® Mice |
MTB Web Interface
Search for Mammary gland
adenocarcinomas with
metastases to the lung &
pathology images available
JAX® Mice | 51
MTB Mammary Adenocarcinomas
with Metastases to Lungs + Images
JAX® Mice | 52
Mammary Gland Images for
FVB/N-Tg(MMTVneu)202Mul/J
JAX® Mice | 53
Mouse Tumor Biology (MTB) Database PDX Models Search
54
http://tumor.informatics.jax.org
JAX® Mice |
PDX Models Search
55 JAX® Mice | tumor.informatics.jax.org/mtbwi/pdxSearch.do
Primary tumor site
Diagnosis
Gene Variants
Gene Expression Chart
Gene Amplification/Deletion (CNVs)
56
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![Whole transcriptome profiling of patient-derived xenograft ...eprints.whiterose.ac.uk/96695/1/WRRO_96695.pdf · xenograft models or specific cancer type [8–9]. In this paper, we](https://static.fdocuments.us/doc/165x107/5f0337437e708231d4081c1a/whole-transcriptome-profiling-of-patient-derived-xenograft-xenograft-models.jpg)
