In vivo genome editing and organoid transplantation models of colorectal cancer and metastasis

Roper, J., Tammela, T., Cetinbas, N. M., Akkad, A., Roghanian, A., Rickelt, S., Almeqdadi, M., Wu, K., Oberli, M. A., Sanchez-Rivera, F., Park, Y. K., Liang, X., Eng, G., Taylor, M. S., Azimi, R., Kedrin, D., Neupane, R., Beyaz, S., Sicinska, E. T., Suarez, Y., Yoo, J., Chen, L., Zukerberg, L., Katajisto, P., Deshpande, V., Bass, A. J., Tsichlis, P. N., Lees, J., Langer, R., Hynes, R. O., Chen, J., Bhutkar, A., Jacks, T., Yilmaz, O. H. (June 2017) In vivo genome editing and organoid transplantation models of colorectal cancer and metastasis. Nat Biotechnol, 35 (6). pp. 569-576. ISSN 1546-1696 (Electronic)1087-0156 (Linking)

URL: https://www.ncbi.nlm.nih.gov/pubmed/28459449
DOI: 10.1038/nbt.3836

Abstract

In vivo interrogation of the function of genes implicated in tumorigenesis is limited by the need to generate and cross germline mutant mice. Here we describe approaches to model colorectal cancer (CRC) and metastasis, which rely on in situ gene editing and orthotopic organoid transplantation in mice without cancer-predisposing mutations. Autochthonous tumor formation is induced by CRISPR-Cas9-based editing of the Apc and Trp53 tumor suppressor genes in colon epithelial cells and by orthotopic transplantation of Apc-edited colon organoids. ApcDelta/Delta;Kras(G12D/+);Trp53Delta/Delta (AKP) mouse colon organoids and human CRC organoids engraft in the distal colon and metastasize to the liver. Finally, we apply the orthotopic transplantation model to characterize the clonal dynamics of Lgr5(+) stem cells and demonstrate sequential activation of an oncogene in established colon adenomas. These experimental systems enable rapid in vivo characterization of cancer-associated genes and reproduce the entire spectrum of tumor progression and metastasis.

Item Type: Paper
Uncontrolled Keywords: Animals Carcinogenesis/genetics Cell Line, Tumor Clustered Regularly Interspaced Short Palindromic Repeats/genetics Colorectal Neoplasms/*genetics *Disease Models, Animal Female Gene Editing/*methods Genes, Neoplasm/*genetics Liver Neoplasms/*genetics/*secondary Male Mice Mice, Transgenic Neoplasm Metastasis Organ Transplantation/*methods
Subjects: Investigative techniques and equipment > cell culture > cancer organoids
diseases & disorders > cancer > cancer types > colon cancer
diseases & disorders > cancer > cancer types > colon cancer

diseases & disorders > cancer > metastasis
CSHL Authors:
Communities: CSHL labs > Beyaz lab
Depositing User: Matt Covey
Date: June 2017
Date Deposited: 01 Feb 2018 21:35
Last Modified: 01 Feb 2018 21:35
PMCID: PMC5462879
Related URLs:
URI: http://repository.cshl.edu/id/eprint/36055

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