Download Modeling Human Cancer in Mice

Modeling Human
Cancer in Mice
40x
10x
Presented by:
Hosted by Charles River, exclusive
distributor of JAX® Mice in 27 European
Countries
Brian W. Soper, PhD
Technical Information
Services
The Jackson Laboratory’s Mission
Performing Research
Investigating genetics and biology of human disease
Providing Resources
JAX® Mice Clinical & Research Services, bioinformatics data,
technical publications and more…
Educating Scientists
World-class courses, internships and other programs
www.jax.org/courses |
2
® Mice
JAX
JAX® Mice
TheGold
Gold Standard
Standard for
The
forBiomedical
BiomedicalResearch
Research
NIH funded resource
>7,000 strains and growing
o 2.7 million mice shipped annually
Unsurpassed genetic quality & animal health
Best characterized & referenced ~100 new pubs/week
Common inbred strains (C57BL/6J, BALB/cJ, DBA/2J) support
development/collection of specialty strains and other valuable
community research resources
JAX® Mice |
3
Online Resources to Expedite
Research
JAX® Mice Database
www.jax.org/jaxmice
www.jax.org/jaxmice/support/techsupport-index |
4
Online Resources to Expedite
Research
JAX® Mice Database
www.jax.org/jaxmice
Mouse Genome Informatics
www.informatics.jax.org
www.jax.org/jaxmice/support/techsupport-index |
5
Online Resources to Expedite
Research
JAX® Mice Database
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 |
6
ACS Cancer Incidence USA
Statistics through 2011
Prostate
Rate per 100,000
Breast
Lung
Lung
80
80
60
60
40
40
20
20
1970
2011
1970
Females
2011
Males
www.cancer.org/research/cancerfactsfigures
JAX® Mice |
7
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
Before treatment
After treatment
Gulliya KS et al. 1994. Cancer 74(6):1725-32. PMID:8082074
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
JAX® Mice |
17
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
Human SK-BR-3 Breast Cancer Cells
Shin S et al. 2011. Mol Cancer Ther 10:603-614. PMID:21393427
JAX® Mice |
21
BALB/c-scid
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
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
Arrows point to
neural rosettes
Werbowetski-Ogilvie TE et al. 2012. Stem Cells 30(3):392-404 PMID:22213600
JAX® Mice |
26
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
Cell line:
o A375 human malignant melanoma
o Luciferase tagged
o Injected I.V.
Carreno et al. 2008. Clin Cancer Res 15(10):3277-86. PMID:19447870
JAX® Mice |
32
NSG Mice Preserve Patient Tumor
Characteristics
Cancer
Patient’s Lung
Tumor
Fragment
NSG
H&E
C.B-17
scid
H&E
Human cells
(anti-HLA)
Mouse blood cells
(anti-mCD45)
Simpson-Abelson MR et al. 2008.J Immunol 180(10):7009-18. PMID:18453623
JAX® Mice |
33
Engrafted
tumor
Patient tumor
NSG Mice Preserve Patient Tumor
Characteristics
H&E
Cytokeratin
Ki67
Simpson-Abelson MR et al. 2008.J Immunol 180(10):7009-18. PMID:18453623
JAX® Mice |
34
JAX Patient-Derived Xenograft
(PDX) Program
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
Access supporting data at
www.tumor.informatics.jax.org/mtbwi/pdxSearch.do
CNV array analysis
In Vivo Pharmacology Services |
35
JAX Patient-Derived Xenograft
(PDX) Program
In Vivo Pharmacology Services |
36
Estrogen Receptor Positive
Breast Carcinoma
TM00386 (ER+/PR+/Her2-)
TM00386 PT
TM00386-294 P0
TM00386-294 P1
PT= Patient Tumor; P0 = first patient derived xenograft (PDX) generation in mouse;
P1 = 2nd generation
NSG supports growth of ER+ breast tumors in combination with estrogen pellets
Tumors retain “organoid” growth pattern
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)
FVB/N Characteristics:
B6 Characteristics:
100% incidence
100% incidence
o Females: median age 53 days
o Pregnancy accelerates
o Females: median age 92 days
tumorigenesis
Metastasize to the lung
Metastasize to the lung
o 80-94% of females
o Reduced compared to FVB/N
background
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
Mammary Cancer Model
STOCK Trp53tm1Brd Brca1tm1Aash Tg(LGB-cre)74Acl/J
(012620)
High Grade Invasive
Ductal Carcinoma
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
Nuclear
Pleomorphism
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
JAX® Mice |
40
Early-Stage Lung Cancer Model
B6.129S4-Krastm4Tyj/J (008179)
5x105 pfu @ 6wks
5x106 pfu @ 6wks
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.
Prostate Cancer Models
TRAMP strains
C57BL/6-Tg(TRAMP)824Ng/J (003135)
(C57BL/6-Tg(TRAMP)8247Ng/J X FVB/NJ)F1/J (008215)
TRansgenic Adenocarcinoma Mouse Prostate
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
Hyperplasia
(~10 weeks)
Adenocarcinoma
(~18wks)
About 28 weeks of age
o SV40 Tumor-antigen
o Variable tumor latency
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
JAX® Mice |
42
Inducible Prostate Cancer Models
FVB;B6-Tg(Pbsn-cre)20Fwan/J (023325)
Probasin cre (Pbsn-cre) alone does not develop tumors
Pbsn-cre::KrasG12D/+::Scribfl/fl generates invasive carcinoma (~400 days)
Publication involving:
B6.129S4-Krastm4Tyj/J (008179)
Scrib loxp-flanked mice
PIN = prostate intraepithelial neoplasia
Pearson HB et al. 2011. J Clin Invest 121(11):4257-67. PMID: 21965329
43
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
Normal intestinal tract
Females unhealthy
o Severely anemic & die by 4 months
o 12% mammary tumor incidence
Homozygous lethal e7.0
Multiple Adenomas
(131 days)
Moser AR et al. 1990. Science 247:322-4. PMID:2296722
Su LK et al. 1992. Science 256:1114. PMID:1350108
JAX® Mice |
44
Melanoma Model
STOCK Braftm1Mmcm Ptentm1Hwu
Tg(Tyr-cre/ERT2)13Bos/BosJ (013590)
Skin
Floxed conversion to BrafV600E
Floxed Pten KO
Inducible melanomas in 3-4 weeks
25% spontaneous melanomas in 2-12 mo.
Lung
Dankort et al. 2009. Nature Genetics 41(5):544-52. PMID:19282848
Damsky et al. 2011. Cancer Cell 20(6):741-54. PMID:22172720
JAX® Mice |
45
Pancreatic Cancer Modeling
B6.FVB-Tg(Pdx1-cre)6Tuv/J (014647)
Pdx1 promoter; pancreatic and duodenal homeobox 1
Triple compound mutant mice develop
pancreatic ductal adenocarcinoma
Recent publication involving:
C;129S4-Ptentm1Hwu/J (004597)
B6.129S4-Krastm4Tyj/J (008179)
Hill et al. 2010. Cancer Res 10(18):7114-24. PMID:20807812
JAX® Mice |
46
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
http://tumor.informatics.jax.org
JAX® Mice |
50
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
http://tumor.informatics.jax.org
JAX® Mice |
54
PDX Models Search
Primary tumor site
Diagnosis
Gene Variants
Gene Expression Chart
Gene Amplification/Deletion (CNVs)
JAX®
Mice |
tumor.informatics.jax.org/mtbwi/pdxSearch.do
55
Thank you!
Charles River is the
Exclusive Distributor of
JAX® Mice in 27
European Countries*
Contact Charles River:
[email protected]
JAX® Mice, Clinical &
Research Services
www.jax.org
*Albania, Austria, Belgium, BosniaHerzegovina, Bulgaria, Croatia, Czech
Republic, Denmark, Finland, France,
Germany, Hungary, Ireland, Italy,
Luxembourg, Macedonia, Montenegro,
Netherlands, Norway, Poland, Portugal,
Serbia, Slovenia, Spain, Switzerland, Sweden,
56
and the United Kingdom