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Comprehensive models of human primary and metastatic colorectal tumors in immunodeficient and immunocompetent mice by chemokine targeting

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Abstract

Current orthotopic xenograft models of human colorectal cancer (CRC) require surgery and do not robustly form metastases in the liver, the most common site clinically. CCR9 traffics lymphocytes to intestine and colorectum. We engineered use of the chemokine receptor CCR9 in CRC cell lines and patient-derived cells to create primary gastrointestinal (GI) tumors in immunodeficient mice by tail-vein injection rather than surgery. The tumors metastasize inducibly and robustly to the liver. Metastases have higher DKK4 and NOTCH signaling levels and are more chemoresistant than paired subcutaneous xenografts. Using this approach, we generated 17 chemokine-targeted mouse models (CTMMs) that recapitulate the majority of common human somatic CRC mutations. We also show that primary tumors can be modeled in immunocompetent mice by microinjecting CCR9-expressing cancer cell lines into early-stage mouse blastocysts, which induces central immune tolerance. We expect that CTMMs will facilitate investigation of the biology of CRC metastasis and drug screening.

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Figure 1: Modeling primary human CRC recurrent mutations in mice without survival surgery.
Figure 2: Human primary CRC-immunoproficient mouse host chimeras.
Figure 3: Sequential human CRC primary GI tumor-metastasis formation.
Figure 4: Increased chemoresistance of human CRC cells in liver versus orthotopic or subcutaneous xenografts.

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Acknowledgements

We thank other members of the Lipkin laboratory and Shen Laboratory for helpful discussions. We also thank the Cornell BRC imaging and Stem Cell and Transgenic Core Facilities. This work was supported by the National Science Foundation (NSF) 1137269, NSF 1106153, NSF GRFP-2011131053, National Institutes of Health (NIH) R01 GM095990, CA098626, NIH 2UL1-RR024996, NIH UH2TR000516, Defense Advanced Research Projects Agency 19-1091726, and generous donations by Matthew Bell and Fred Greenberg.

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Contributions

H.J.C., X.S. and S.M.L. proposed the concept, designed the experiments and co-wrote the manuscript; H.J.C., Z.H., M.A., N.R., Joyce Chen, P.B., L.W., Y.X., Jonlin Chen and M.S. performed the experiments; J.S., K.Y.C. and Z.H.G. contributed to bioinformatics analyses; H.H.J., R.M., C.A. and W.E. performed blastocyst injections; N.R., D.J.J., Jiahn Choi, P.G. and N.N. performed multiphoton imaging; J.M. and G.N. provided tissue samples; R.L. performed assays for detecting immune cells; X.K.Z. performed statistical analysis; R.E. and A.D.M. performed histopathology analyses.

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Correspondence to Xiling Shen or Steven M Lipkin.

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The authors declare no competing financial interests.

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Supplementary Figures 1–12, Supplementary Tables 1–3, Supplementary Methods (PDF 14691 kb)

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Chen, H., Sun, J., Huang, Z. et al. Comprehensive models of human primary and metastatic colorectal tumors in immunodeficient and immunocompetent mice by chemokine targeting. Nat Biotechnol 33, 656–660 (2015). https://doi.org/10.1038/nbt.3239

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