Upcoming JAX Webinars™
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February 20, 2014, 1pm ET, USA
Innovative Preclinical Research Using Humanized NSG Mice
February 27, 2014, 1pm ET, USA
Tips for the Care and Housing of Immune Deficient Mice
March 6, 2014, 1pm ET, USA
All You Need is Love: Reproductive Biology of Mice
March 13, 2014, 1pm ET, USA
S.O.S. Colony Adrift: Protecting Mouse Colonies from Genetic Drift
March 20, 2014, 1pm ET, USA
Choosing and Using Mouse Models of Huntington’s Disease
www.jax.org/jaxmice/webinar |
1
Make Your Life in the Lab Easier
MouseClique Blog
mouseclique.jax.org |
2
Cutting Edge
Human Disease
Modeling Using
The NSG Mouse
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,
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World-class courses, internships and other programs
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® Mice
JAX
JAX® Mice
TheGold
Gold Standard
Standard for
The
forBiomedical
BiomedicalResearch
Research
NIH funded resource
>7,000 strains and growing
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
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Online Resources to Expedite
Research
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Online Resources to Expedite
Research
JAX® Mice Database
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www.informatics.jax.org
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Online Resources to Expedite
Research
JAX® Mice Database
www.jax.org/jaxmice/query
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 |
8
Cutting Edge Human Disease
Modeling: The NSG Mouse
Why NSG is so valuable
How NSG elevates the quality
of human disease models
o Solid and hematological cancers
o Human immunity
o Transplantation research
What the future holds for
advanced disease models
How to find more information
to support your research
JAX® Mice |
9
Cutting Edge Human Disease
Modeling: The NSG Mouse
Why NSG is so valuable
How NSG elevates the quality of
human disease models
o Solid and hematological cancers
o Human immunity
o Transplantation research
What the future holds for
advanced disease models
How to find more information to
support your research
JAX® Mice |
10
The Immune System in Mice
JAX® Mice |
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NOD scid gamma (NSG)
Strain nomenclature: NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ
Stock Number: 005557
Additional resources: NSG Web Resources
www.jax.org/jaxmice/research/immunology/005557.html
JAX® Mice |
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NSG:
A Highly Immunodeficient Mouse
NOD background contributes:
Absence of hemolytic complement
Reduced dendritic and NK cell function
Defective macrophages
Optimal human hematopoietic stem cell engraftment (Sirpa
allele)
JAX® Mice |
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NSG:
A Highly Immunodeficient Mouse
JAX® Mice |
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NSG:
A Highly Immunodeficient Mouse
NOD background contributes:
Absence of hemolytic complement
Reduced dendritic and NK cell function
Defective macrophages
Optimal human hematopoietic stem cell engraftment
(Sirpa allele)
scid mutation prevents development of mature T and B cells
JAX® Mice |
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NSG:
A Highly Immunodeficient Mouse
JAX® Mice |
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NSG:
A Highly Immunodeficient Mouse
NOD background contributes:
Absence of hemolytic complement
Reduced dendritic and NK cell function
Defective macrophages
Optimal human hematopoietic stem cell engraftment
(Sirpa allele)
scid mutation prevents development of mature T and B cells
gamma chain (Il2rg) knockout eliminates signaling from 6 distinct
interleukins and blocks NK cell development
JAX® Mice |
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Gamma Chain Knockout (Il2rgtm1Wjl)
Complete deficiency of the interleukin 2 receptor, gamma chain
Common subunit found in multiple cytokine cell signaling
receptors
IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21 are key signaling molecules
in function and maturation of T, B, NK, neutrophils and dendritic
cells
Rochman Y et al. 2009. Nat Rev Immunol 9(7):480-90. PMID:19543225
JAX® Mice |
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NSG:
A Highly Immunodeficient Mouse
JAX® Mice |
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NSG:
A Highly Immunodeficient Mouse
Longer lifespan (> 16 months) than NOD-scid (~ 9
months)
scid side effects: Radiation sensitivity; genotoxic drugs
can have higher toxicity
A platform for developing refined models (transgenic
and knockout)
JAX® Mice |
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The Most Useful and Versatile
Immunodeficient Mouse
Research applications:
Primary tumor engraftment
Cancer stem cells
Human hematopoiesis
Humanized mice
Infectious disease
Transplantation research
JAX® Mice |
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Evaluating Cancer Therapies
Using Xenografts in Nude Mice
Before treatment
After treatment
Gulliya et al. 1994. Cancer 74:1725-1732. PMID:8082074
JAX® Mice |
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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 |
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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 |
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JAX Patient-Derived Xenograft
(PDX) Program
All-comers patient-derived tumor bank in NSG mice
Clinical information
o Tumor type, grade and markers (if known)
o Treatment history
Histology
Gene expression array and CNV array analysis
In Vivo Pharmacology Services |
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JAX Patient-Derived Xenograft
(PDX) Program
In Vivo Pharmacology Services |
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Established PDX Models
(306 to date)
Tumor type
Number of models
Appendiceal
1
Bladder
20
Brain
33
Breast
20
Colorectal
36
Endometrial
4
Gastric/GIST
8
Hematology
7
Hepatocellular
1
Kidney
57
Lung
48
Ovary
12
Pancreas
18
Prostate
3
Sarcoma
11
Skin
8
In Vivo Pharmacology Services |
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Efficient Modeling of ER+
Breast Cancer
BR0555F
BR0555F-294
BR0555F-294
Patient Tumor
First generation in NSG
Second generation in
NSG
NSG supports growth of ER+ breast tumors
Tumors retain “organoid” growth pattern
In Vivo Pharmacology Services |
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Orthotopic Bladder Cancer
PDX Model
Fidelity during tumor passaging
(subcutaneous)
Orthotopic model
Lin TY et al. 2012. Nanomedicine (Lond).[Epub ahead of print] PMID:23199207
In Vivo Pharmacology Services |
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Efficacy in Bladder Cancer
PDX Model
Tumor-Targeting Micelles Prevent
Tumor Growth…
…and Improve Survival
Lin TY et al. 2012. Nanomedicine (Lond).[Epub ahead of print] PMID:23199207
In Vivo Pharmacology Services |
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The Most Permissive Host for
Cancer Stem Cells
Single human melanoma cell
NSG
Quintana E et al. 2008. Nature 456(7222):593-8. PMID:19052619
In Vivo Pharmacology Services |
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Improved Melanoma Metastasis
Modeling
Cell line: A375 (RhoC, Luciferase) injected I.V.
Carreno et al. 2008.
Clin Cancer Res 15(10):3277-86. PMID:19447870
In Vivo Pharmacology Services |
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Improved Cell Line Engraftment
with NSG
Carreno et al. 2009. Clin Cancer Res15(10):3277-86. PMID:19447870
In Vivo Pharmacology Services |
33
Cutting Edge Human Disease
Modeling: The NSG Mouse
Why NSG is so valuable
How NSG elevates the quality
of human disease models
o Solid and hematological cancers
o Human immunity
o Transplantation research
What the future holds for
advanced disease models
How to find more information to
support your research
JAX® Mice |
34
Improved Leukemia Cell
Engraftment
NSG
NOD-scid B2m
NOD-scid
Cells: 697 cells (ALL-B)
Route: I.P.
Age of mice: 6-8 weeks
Agliano A et al. 2008. Int J Cancer 123(9):2222-7. PMID:18688847
JAX® Mice |
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Effective Modeling of PatientDerived ALL (T & B Cell)
Cells: Unsorted patient-derived T or B cell ALL
Route: Tail vein
Age of mice: 6-8 weeks
Patient samples engrafted
at lower doses and with
faster kinetics in NSG
Diamanti P et al. 2011. Leukemia Feb;26(2):376-80. PMID:21860430
JAX® Mice |
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Non-Invasive Imaging of ALL PDX
Lentivirus can be
used to deliver
reporter genes to
ALL PDX
Noninvasive,
whole-body
imaging permits
tracking of disease
progression
Secondary
passaging is
possible
Bomken S et al. 2012. Leukemia [Epub ahead of print]. PMID:22858908
JAX® Mice |
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NSG Efficiently Models Patient AML
Samples from different patients showed improved engraftment versus other models (NSB)
Sanchez PV et al. 2009. Leukemia 23(11):2109-2117. PMID:19626050
JAX® Mice |
38
Cutting Edge Human Disease
Modeling: The NSG Mouse
Why NSG is so valuable
How NSG elevates the quality
of human disease models
o Solid and hematological cancers
o Human immunity
o Transplantation research
What the future holds for
advanced disease models
How to find more information to
support your research
JAX® Mice |
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NSG Supports a Human Immune
System
Most efficient engraftment of human HSCs to date
Permits differentiation of all major cell types:
o Myeloid lineage
Erythrocytes, megakaryocytes, platelets
Monocytes, macrophages, dendritic cells, granulocytes
o Lymphoid lineage
B cells
T cells (CD4+ and CD8+)
NK cells
Some mucosal and adaptive immune function
JAX® Mice |
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NSG Supports a Human Immune
System
NSG is a superior host for human immune
cells when compared to other strains:
1. Any strain expressing the scid mutation alone (NOD-scid,
B6-scid, C.B17-scid)
2. “scid beige”
3. The same mutations—scid (or a Rag1 or Rag2 knockout)
and “gamma”—on other backgrounds
(for example, BALB/c)
Shultz LD et al. 2005. J Immunol 174(10):6477-89. PMID:15879151
Lepus CM et al. 2009. Hum Immunol Oct;70(10):790-802. PMID:19524633
Brehm MA et al. 2010. Clin Immunol 135(1):84-98. PMID:20096637
JAX® Mice |
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Improved WBC Differentiation in
NSG
Stained for human CD45
NSG
NOD-scid
Low mag
High mag
10 weeks post-engraftment
Shultz LD et al. 2005. J Immunol 174(10):6477-89 PMID:15879151
Brehm MA et al. 2010. Clin Immunol 135(1):84-98 PMID:20096637
JAX® Mice |
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NSG: The Most Efficient
Humanized Model
Engraftment Efficiency of Bone Marrow
Strain
Exp 1
Exp 2
Combined
Percentage
NOD/Lt-scid
9/11
5/7
14/18
77.8
NOD/Shi-scid
12/12
3/11
15/23
65.2
NSG
12/12
11/11
23/23
100
NOG
10/12
9/10
19/22
86.4
NSG also had the greatest efficiency in spleen & thymus
Adapted from McDermott SP et al. 2010. Blood. Jul 15;116(2):193-200. PMID:20404133
JAX® Mice |
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Humanized NSG Experimental
Timeline
Whole body
irradiation
Tail vein
injection
Human B
cells appear
Human T
cells appear
8 weeks
11 weeks
15 weeks
24~48 hours3 weeks
Mouse
age
In Vivo Pharmacology Services |
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12 w
Successful Engraftment in
Peripheral Blood
Human
Engrafted NSG
Mouse
Human CD45
Mostly
neutrophils,
monocytes
Mouse CD45
Human white blood cells were abundant
in circulation
Human CD3
Mature human T (CD3+) and B (CD20+)
cells differentiated in the NSG mouse
Human CD20
In Vivo Pharmacology Services |
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Multilineage Differentiation in Bone
Marrow
Human vs Mouse
Human HSC
Human Erythroid
11.2%
hCD34
hCD45
SSC
FSC
0.13%
88.6%
86.1%
mCD45
Human B cells
hCD38
Human APCs
DTH Response in Humanized NSG is
Evidence for Intact T Cell Functions
Sensitization Sensitization Challenge +/- treatment
Day -1
Day 1
Day 2
Day 7
Day 8
Day 9
Day 14
Ear thickness Ear thickness Ear thickness
Ear thickness Ear thickness Ear thickness
Naive
Sensitized with DNFB
Sensitized with Olive Oil
0.30
DNFB + Hydrocortisone
0.25
w
ee
k
1
ho
ur
s
48
24
ho
ur
s
0.20
Ba
se
lin
e
Se
ns
itiz
at
io
n
Ear Measurement (mm)
0.35
In Vivo Pharmacology Services |
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% Viable hCD45+
peripheral cells
Stable Engraftment After
Transportation
In Vivo Pharmacology Services |
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Research Ready Humanized NSG
NSG engrafted with human CD34+ hematopoietic stem cells
Availability of BLT & PBMC engrafted mice
Mice are delivered 12 weeks after engraftment with at least 25%
hCD45+ cells in the peripheral blood
In Vivo Pharmacology Services |
49
Cutting Edge Human Disease
Modeling: The NSG Mouse
Why NSG is so valuable
How NSG elevates the quality
of human disease models
o Solid and hematological cancers
o Human immunity
o Transplantation research
What the future holds for
advanced disease models
How to find more information to
support your research
JAX® Mice |
50
Improved Engraftment with PBMCs
King M et al. 2008. Clin Immunol 126(3):303-14. PMID:18096436
In Vivo Pharmacology Services |
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Improved Engraftment with PBMCs
BRG: BALB/c Rag2, Il2rg
double knockout
Ali N et al. 2012. Plos ONE 7(8):e44219. PMID:22937164
In Vivo Pharmacology Services |
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Transplant Rejection Studies:
Diabetes
Insulin
hCD45
H&E
Diabetic mice: NSG treated with STZ
Transplanted with human islets, +/PBMCs
32 days post-engraft.
No PBMC
+ PBMC
King M et al. 2008. Clin Immunol 126(3):303-14. PMID:18096436
JAX® Mice |
53
Cutting Edge Human Disease
Modeling: The NSG Mouse
Why NSG is so valuable
How NSG elevates the quality of
human disease models
o Solid and hematological cancers
o Human immunity
o Transplantation research
What the future holds for
advanced disease models
How to find more information to
support your research
JAX® Mice |
54
NOD Rag1 Gamma (NRG):
A Radiation Resistant Alternative to NSG
Nomenclature: NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl/SzJ
Stock Number: (007799)
Webpage: www.jax.org/jaxmice/strain/007799
Severe defects in both innate and adaptive immunity
Hematopoietic stem cell engraftment comparable to NSG
Radiation resistant
Emerging model
Pearson T et al. 2008. Clin Exp Immunol 154(2): 270-84. PMID:18785974
JAX® Mice |
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New NSG Model Innovations
PBMC Engraftment
NOD.Cg-Prkdcscid B2mtm1Unc Il2rgtm1Wjl/SzJ (010636)
o MHC class I deficient NSG strain with enhanced PBMC
engraftment
HLA Class I Transgenic
NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(HLA/H2D/B2M)1Dvs/SzJ (014570)
o HLA Class I heavy and light chains (A2.1 haplotype)
NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(HLA-A2.1)1Enge/SzJ
(009617)
o Expresses HLA Class I transgene (A2.1 haplotype)
JAX® Mice |
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New NSG Model Innovations
HLA Class II Transgenic
NOD.Cg-Prkdcscid Il2rgtm1Wjl H2-Ab1tm1Gru Tg(HLADRB1)31Dmz/SzJ (017637)
o Lacks mouse MHC class II, expresses chimeric HLA class II
DR4 transgene
NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(HLA-DRA*0101,HLA
DRB1*0101)1Dmz/GckRoly (012479)
o NSG strain expresses HLA class II transgene (DR1
haplotype)
NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl Tg(HLA-DRA,HLADRB1*0401)39-2Kito/ScasJ (017914)
o “DRAG” strain with HLA class II transgene (DR4; HLADRA/HLA-DRB1*0401)
JAX® Mice |
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New NSG Model Innovations
Cytokine Transgenic
NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(PGK1KITLG*220)441Daw/SzJ (017830)
o Kit ligand (SCF) transgenic that does not require preconditioning
irradiation
NOD.Cg-Prkdcscid Il2rgtm1Wjl
Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ (013062)
o Expresses human IL-3, CSF2, Kit ligand
o Supports human myeloid cells (CD34-humanized immune system;
acute myeloid leukemia)
Cancer Research
NOD.Cg-Prkdcscid Il2rgtm1Wjl Hprtb-m3/EshJ (012480)
o Hprt-null NSG mouse
o Addition of HAT medium depletes mouse cells to enable isolation of
human cells
JAX® Mice |
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Developing New NSG Strains
Starting with an existing
transgene or knockout…
Requires a backcross to
NOD/ShiLtJ (001976), then
intercross with NSG
Speed congenics protocols
reduce time in half
Engineering new mutations…
NOD ES cell line available from
The Jackson Laboratory
Landel and colleagues isolated
germline competent ES cells
from NSG
Landel CP et al. 2013. Transgenic Res 22(1):179-85. PMID:22767020
Breeding & Rederivation Services |
59
Cutting Edge Human Disease
Modeling: The NSG Mouse
Why NSG is so valuable
How NSG elevates the quality of
human disease models
o Solid and hematological cancers
o Human immunity
o Transplantation research
What the future holds for
advanced disease models
How to find more information
to support your research
JAX® Mice |
60
Online NSG Resources
www.jax.org/jaxmice/research/immunology/005557
JAX® Mice |
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Categorized & Annotated NSG
References List
IMMUNOLOGY
STEM CELLS / REGENERATIVE MEDICINE
Hematopoiesis and stem cell engraftment
T cell function / B cell function / Myeloerythroid lineages
NSG as a superior host for human immune system
components
NSG strains expressing human cytokine transgenes
NSG strains expressing HLA transgenes
Reviews and protocols
Transplantation research / graft-versus-host disease
(GVHD) / PBMC engraftment
Vaccines
Autoimmunity
INFECTIOUS DISEASE
Malaria
Cytomegalovirus
Dengue virus
Epstein-Barr
HIV modeling & therapeutics
Measles
Sepsis
Typhoid fever
Varicella-zoster virus
DIABETES
SOLID TUMORS
Breast cancer
Brain cancer
Cancer stem cells
Colon cancer
Esophageal cancer
Head and neck cancer
Hepatoblastoma
Hepatocellular carcinoma
Lung cancer
Immunotherapy
Melanoma / metastasis
Ovarian cancer
Prostate cancer
Sarcoma
HEMATOLOGICAL CANCERS
AML
Lymphoma
ALL
Mastocytoma
CML
Multiple myeloma
CLL
Chronic lymphocytic leukemia
www.jax.org/jaxmice./research/immunology/005557
JAX® Mice |
63
Thank you!
In need of mouse breeding and colony
management expertise to advance your
research?
Contact your regional representative today
www.jax.org/jaxmice/support/regionalcontacts
Contact technical
supportwww.jax.org/jaxmice/support/techsupportindex.html
JAX® Mice, Clinical & Research Services
[email protected] • www.jax.org/jaxmice
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