Sequential cancer mutations in cultured human intestinal stem cells
Publication date
2015-05-07
Authors
Drost, Jarno
van Jaarsveld, Richard H
Ponsioen, Bas
Zimberlin, Cheryl
van Boxtel, Ruben
Buijs, Arjan
Sachs, Norman
Overmeer, Rene M.
Offerhaus, G Johan
Begthel, Harry
Editors
Advisors
Supervisors
Document Type
Article
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taverne
Abstract
Crypt stem cells represent the cells of origin for intestinal neoplasia. Both mouse and human intestinal stem cells can be cultured in medium containing the stem-cell-niche factors WNT, R-spondin, epidermal growth factor (EGF) and noggin over long time periods as epithelial organoids that remain genetically and phenotypically stable. Here we utilize CRISPR/Cas9 technology for targeted gene modification of four of the most commonly mutated colorectal cancer genes (APC, P53 (also known as TP53), KRAS and SMAD4) in cultured human intestinal stem cells. Mutant organoids can be selected by removing individual growth factors from the culture medium. Quadruple mutants grow independently of all stem-cell-niche factors and tolerate the presence of the P53 stabilizer nutlin-3. Upon xenotransplantation into mice, quadruple mutants grow as tumours with features of invasive carcinoma. Finally, combined loss of APC and P53 is sufficient for the appearance of extensive aneuploidy, a hallmark of tumour progression.
Keywords
IN-VITRO EXPANSION, COLORECTAL-CANCER, STRUCTURAL BASIS, HUMAN COLON, ORGANOIDS, ADENOMAS, TRANSFORMATION, EPITHELIUM, REVEALS, SMAD4, Taverne, Journal Article, Research Support, Non-U.S. Gov't
Citation
Drost, J, van Jaarsveld, R H, Ponsioen, B, Zimberlin, C, van Boxtel, R, Buijs, A, Sachs, N, Overmeer, R M, Offerhaus, G J, Begthel, H, Korving, J, van de Wetering, M, Schwank, G, Logtenberg, M, Cuppen, E, Snippert, H J, Medema, J P, Kops, G J P L & Clevers, H 2015, 'Sequential cancer mutations in cultured human intestinal stem cells', Nature, vol. 521, no. 7550, pp. 43-U329. https://doi.org/10.1038/nature14415