Sequential cancer mutations in cultured human intestinal stem cells

Jarno Drost; Richard H. Van Jaarsveld; Bas Ponsioen; Cheryl Zimberlin; Ruben Van Boxtel; Arjan Buijs; Norman Sachs; René M. Overmeer; G. Johan Offerhaus; Harry Begthel; Jeroen Korving; Marc Van De Wetering; Gerald Schwank; Meike Logtenberg; Edwin Cuppen; Hugo J. Snippert; Jan Paul Medema; Geert J P L Kops; Hans Clevers

doi:10.1038/nature14415

Sequential cancer mutations in cultured human intestinal stem cells

Jarno Drost, Richard H. Van Jaarsveld, Bas Ponsioen, Cheryl Zimberlin, Ruben Van Boxtel, Arjan Buijs, Norman Sachs, René M. Overmeer, G. Johan Offerhaus, Harry Begthel, Jeroen Korving, Marc Van De Wetering, Gerald Schwank, Meike Logtenberg, Edwin Cuppen, Hugo J. Snippert, Jan Paul Medema, Geert J P L Kops, Hans Clevers

Research output: Contribution to journal › Article › peer-review

481 Scopus citations

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.

Original language	English (US)
Pages (from-to)	43-47
Number of pages	5
Journal	Nature
Volume	521
Issue number	7550
DOIs	https://doi.org/10.1038/nature14415
State	Published - May 7 2015
Externally published	Yes

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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, 521(7550), 43-47. https://doi.org/10.1038/nature14415

Drost, J, Van Jaarsveld, RH, Ponsioen, B, Zimberlin, C, Van Boxtel, R, Buijs, A, Sachs, N, Overmeer, RM, Offerhaus, GJ, Begthel, H, Korving, J, Van De Wetering, M, Schwank, G, Logtenberg, M, Cuppen, E, Snippert, HJ, Medema, JP, Kops, GJPL & Clevers, H 2015, 'Sequential cancer mutations in cultured human intestinal stem cells', Nature, vol. 521, no. 7550, pp. 43-47. https://doi.org/10.1038/nature14415

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title = "Sequential cancer mutations in cultured human intestinal stem cells",

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.",

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AU - Drost, Jarno

AU - Van Jaarsveld, Richard H.

AU - Ponsioen, Bas

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AU - Van Boxtel, Ruben

AU - Buijs, Arjan

AU - Sachs, Norman

AU - Overmeer, René M.

AU - Offerhaus, G. Johan

AU - Begthel, Harry

AU - Korving, Jeroen

AU - Van De Wetering, Marc

AU - Schwank, Gerald

AU - Logtenberg, Meike

AU - Cuppen, Edwin

AU - Snippert, Hugo J.

AU - Medema, Jan Paul

AU - Kops, Geert J P L

AU - Clevers, Hans

PY - 2015/5/7

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N2 - 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.

AB - 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.

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Sequential cancer mutations in cultured human intestinal stem cells

Abstract

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