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Supplementary Tables
Table S1. Characteristics of Immunodeficient Mouse Strains
Common Strain Name
Common Abbreviation
Immunological Characteristics Relative EngraftmentSolid PDX Leukemia
HIS*
Commercial Vendor
Athymic nude mice Nude, BALB/c nude
Defective in T (function); retains B, macrophage, NK, dendritic cells
+ - + Multiple
C.B17-scid SCID Defective in T, B; retains NK and macrophage, dendritic activity
+ +/-+/-
Multiple
NOD-scid NOD NOD-SCID Defective in T, B; highly reduced NK and macrophage, dendritic activity
++ + + Multiple
NOD.cg-Prkdcscid
IL2rgtm1SugNOG Defective in T, B and NK cells; defects in
innate immune cell development; lacks IL2rg# intracytoplasmic domain; cytokine
binding but no signaling
+++ ++ +++
Taconic Biosciences (CIEA NOG mouse)
NOD.Cg-Prkdcscid
IL2rgtm1WjlNSG™ Defective in T, B and NK cells; defects in
innate immune cell development; IL2rg null mutation; no cytokine binding or
signaling
+++ ++ +++
The Jackson Laboratory
Note: *HIS: Human immune system; #IL-2 receptor common gamma chain
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Table S2. Examples of Syngeneic Mouse Tumor Cell LinesCancer Type Mouse Tumor Cell
LineBladder MBT-2
Breast4T1
EMT6JC
ColonColon26CT-26MC38
Fibrosarcoma WEHI-164Kidney Renca
Leukemia C1498L1210
Liver H22
Lung KLN205LL/2
Lymphoma
A20E.G7-OVA
EL4L15178-RP388D1
Mastocytoma P815
MelanomaB16-BL6B16-F10
S91Myeloma MPC-11Neuroblastoma Neuro-2aPancreatic Pan02Plasmacytoma J558Prostate RM-1
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Table S3. Examples of GEMM and Derived Allografts
Allograft ID Cancer Type Engineered Mutations/Carcinogen Strain Background
mPR6003 Prostate TRAMP (Pbsn-SV40T TG) C57BL/6
mBR6004 Breast MMTV-PyVT TG(1) FVB/N
mSK6005 Skin squamous cell carcinoma ApcMin/+ C57BL/6
mLY6043 B cell lymphoma IgH-Myc TG (EµMyc ¿ C57BL/6
mSA9003 Sarcoma P53-/- C57BL/6
mHN6041 HNSCC KRAS (G12D); P53-/- C57BL/6
mLU6042, mLU6044, mLU6045
Lung KRAS (G12D); P53-/- C57BL/6
mCR6046 Colon KRAS (G12D); P53-/- C57BL/6
mLI6047 Liver KRAS (G12D); P53-/- C57BL/6
Pancreatic KRAS (G12D); P53-/- C57BL/6
Prostate KRAS (G12D); PTENFlox/Flox C57BL/6
Ovarian KRAS (G12D); PTENFlox/Flox C57BL/6
Bladder KRAS (G12D); PTENFlox/Flox C57BL/6
Lung KRAS (G12D); PTENFlox/Flox C57BL/6
Glioblastoma KRAS s (G12D); PTENFlox/Flox C57BL/6
Bladder PTENFlox/Flox; P53-/- C57BL/6
Prostate PTENFlox/Flox; P53-/- C57BL/6
Pancreatic KRAS (G12D); P16-/- C57BL/6
Lung KRAS (G12D); P16-/- C57BL/6
mNE6014 Neuroendocrine tumor N/A NSG (NOD)
m6032 Cutaneous tumor N/A BALB/c nude
mLE6026 Leukemia N/A NSG (NOD)
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Allograft ID Cancer Type Engineered Mutations/Carcinogen Strain Background
NASH-HCC Streptozotocin and high-fat diet C57BL/6
HCC Diethylnitrosamine and phenobarbital C3H/He
Lung Urethane BALB/c; C3H/He; A/J
AML NRAS (G12D); AML1-ETO C57BL/6
AML NRAS (G12D); MLL-ENL C57BL/6
Bladder P53 cKO; PTEN cKO C57BL/6
Breast P53 cKO; BRCA1 cKO; MMTV-Cre C57BL/6
Breast P53 cKO; SMAD4 cKO; CDH1 cKO; MMTV-Cre C57BL/6
Gastric P53 cKO; SMAD4 cKO; CDH1 cKO C57BL/6
Lung KRAS (G12D); P53 cKO; LKB1 cKO C57BL/6
Pancreatic KRAS (G12D); P53 (R172H); Pdx1-Cre C57BL/6
Pancreatic KRAS (G12D); SMAD5 cKO; Pdx1-Cre C57BL/6
Prostate BRAF (V600E); PTEN cKO; Pbsn-Cre C57BL/6
Prostate PTEN cKO; Smad4 cKO; Pbsn-Cre C57BL/6
Ovarian KRAS (G12D); PTEN cKO C57BL/6
Medullablastoma PTCH1 cKO; P53 cKO C57BL/6
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Supplementary Figures
Figure S1. Histopathology of Pancreatic Cancer. H&E staining – tumor cells (star) and fibrotic stroma (arrow): Pa: original patient; P0: first generation PDX; P1: second generation PDX. CDX: pancreatic cell line derived xenografts.
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Figure S2. Spider Plot of Syngeneic MC38 Growth Inhibition Curves under Anti-PD-1 Antibody Treatment
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Figure S3. Illustration of PD Biomarker Analysis. MC38 tumor treated by anti-mCTLA-antibody: IHC-mCD3 staining of TILs; flow analysis of mCD4/mCD8 of TILs.
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Figure S4. The Concept of Primary Mouse Tumor Homografts
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Figure S5. The Concept of Chimeric GEMM with Humanized Targets. A human target, e.g. human CTLA-4 (Panel A) or PD-1 (Panel B), is knocked in to the mouse, and can interact with human therapeutics.
Figure S5-Panel A
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Figure S5-Panel B
.
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Figure S6. Robust and sustained T lymphopoiesis in peripheral blood (PB) and spleen of Hu-M™ mice. (A) Percentages of huCD45+ (lymphocytes), (B) huCD3+ (T-cells), huCD4+ (TH) and huCD8+ (CTLs) T-cells in the PB of Hu-M mice at 12 and 31-32 weeks post engraftment. (C) Representative dot plot of T-cell subsets in the spleen of Hu-M mice at 25 weeks post engraftment.
Reconstitution of Human Immunity in Immuno-Compromised mice (Hu-M Mice). HuMurine’s Hu-M mice are constructed by
injecting pre-qualified human CD34+ hematopoietic progenitor cells (HPCs) into newborn NOG mice resulting in stable multi-lineage human
hematopoiesis at 10-15 weeks postengraftment. HuMurine (www.humurine.com) has generated over a thousand Hu-M mice with a reproducibly
high and robust engraftment rate. Greater than 80% of injected mice developed humanization levels of >30% as determined by the ratio of human
lymphocytes to murine cells in the peripheral blood at 10-12 weeks postengraftment. The Hu-M mice display human T-cells (CD4+, CD8+), B cells
(CD19+), as well as lower levels of human NK cells (CD56+) and macrophages (CD14+). These mice can sustain enhanced differentiation and
maturation of human CD4+ (TH1 cells) and CD8+ T cytotoxic T-cells (CTLs) for up to 32 weeks postengraftment (as shown in Figure S6). Since
human T-cells clearly mature in the murine Hu-NOG mouse thymus, it has been speculated and proposed that the injected CD34 + cells may
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themselves create a chimeric thymic microenvironment that may allow for some "normal" human T-cell maturation to occur within the humanized
mouse(1,2).
1. Ishikawa, F., Yasukawa, M., Lyons, B., Yoshida, S., Miyamoto, T., Yoshimoto, G., et al. (2005). Development of functional human blood and immune systems in NOD/SCID/IL2 receptor γ chain(null) mice. Blood, 106, 1565-1573.
2. Traggiai, E., Chicha, L., Mazzucchelli, L., Bronz, L., Piffaretti, J. C., Lanzavecchia, A., et al. (2004). Development of a human adaptive immune system in cord blood cell-transplanted mice. Science, 304, 104-107.
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