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

Onderzoeksoutput: Bijdrage aan tijdschrift › Artikel › peer review

888 Citaten (Scopus)

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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.

Originele taal-2	Engels
Pagina's (van-tot)	43-7
Aantal pagina's	5
Tijdschrift	Nature
Volume	521
Nummer van het tijdschrift	7550
DOI's	https://doi.org/10.1038/nature14415
Status	Gepubliceerd - 7 mei 2015
Extern gepubliceerd	Ja

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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), 43-7. https://doi.org/10.1038/nature14415

@article{6729d7b275ad46db842285e65e69e556,

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. ",

keywords = "Aneuploidy, Animals, CRISPR-Cas Systems, Child, Child, Preschool, Colorectal Neoplasms/genetics, Female, Genes, APC, Genes, p53/genetics, Heterografts, Humans, Imidazoles, Intercellular Signaling Peptides and Proteins/metabolism, Intestinal Mucosa/metabolism, Intestines/pathology, Mice, Middle Aged, Mutagenesis, Site-Directed, Mutation/genetics, Neoplasm Invasiveness/genetics, Neoplasm Transplantation, Organoids/metabolism, Piperazines, Proto-Oncogene Proteins p21(ras)/genetics, Smad4 Protein/deficiency, Stem Cell Niche/physiology, Stem Cells/metabolism",

author = "Jarno Drost and \{van Jaarsveld\}, \{Richard H\} and Bas Ponsioen and Cheryl Zimberlin and \{van Boxtel\}, Ruben and Arjan Buijs and Norman Sachs and Overmeer, \{Ren{\'e} M\} and Offerhaus, \{G Johan\} and Harry Begthel and Jeroen Korving and \{van de Wetering\}, Marc and Gerald Schwank and Meike Logtenberg and Edwin Cuppen and Snippert, \{Hugo J\} and Medema, \{Jan Paul\} and Kops, \{Geert J P L\} and Hans Clevers",

year = "2015",

month = may,

day = "7",

doi = "10.1038/nature14415",

language = "English",

volume = "521",

pages = "43--7",

journal = "Nature",

issn = "0028-0836",

publisher = "Nature Research",

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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, nr. 7550, blz. 43-7. https://doi.org/10.1038/nature14415

TY - JOUR

T1 - Sequential cancer mutations in cultured human intestinal stem cells

AU - Drost, Jarno

AU - van Jaarsveld, Richard H

AU - Ponsioen, Bas

AU - Zimberlin, Cheryl

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

Y1 - 2015/5/7

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.

KW - Aneuploidy

KW - Animals

KW - CRISPR-Cas Systems

KW - Child

KW - Child, Preschool

KW - Colorectal Neoplasms/genetics

KW - Female

KW - Genes, APC

KW - Genes, p53/genetics

KW - Heterografts

KW - Humans

KW - Imidazoles

KW - Intercellular Signaling Peptides and Proteins/metabolism

KW - Intestinal Mucosa/metabolism

KW - Intestines/pathology

KW - Mice

KW - Middle Aged

KW - Mutagenesis, Site-Directed

KW - Mutation/genetics

KW - Neoplasm Invasiveness/genetics

KW - Neoplasm Transplantation

KW - Organoids/metabolism

KW - Piperazines

KW - Proto-Oncogene Proteins p21(ras)/genetics

KW - Smad4 Protein/deficiency

KW - Stem Cell Niche/physiology

KW - Stem Cells/metabolism

UR - https://www.scopus.com/pages/publications/84929103936

U2 - 10.1038/nature14415

DO - 10.1038/nature14415

M3 - Article

C2 - 25924068

SN - 0028-0836

VL - 521

SP - 43

EP - 47

JO - Nature

JF - Nature

IS - 7550

ER -

Sequential cancer mutations in cultured human intestinal stem cells

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