Sequential cancer mutations in cultured human intestinal stem cells

被引：833

作者：

Drost, Jarno ^{[1
,2
,3
]}

van Jaarsveld, Richard H. ^{[3
,4
]}

Ponsioen, Bas ^{[3
,4
]}

Zimberlin, Cheryl ^{[3
,5
]}

van Boxtel, Ruben ^{[1
,2
,3
]}

Buijs, Arjan ^{[6
]}

Sachs, Norman ^{[1
,2
,3
]}

Overmeer, Rene M. ^{[3
,4
]}

Offerhaus, G. Johan ^{[7
]}

Begthel, Harry ^{[1
,2
,3
]}

Korving, Jeroen ^{[1
,2
,3
]}

van de Wetering, Marc ^{[1
,2
,3
,8
]}

Schwank, Gerald ^{[1
,2
,3
]}

Logtenberg, Meike ^{[1
,2
,3
]}

Cuppen, Edwin ^{[1
,2
,3
]}

Snippert, Hugo J. ^{[3
,4
]}

Medema, Jan Paul ^{[3
,5
]}

Kops, Geert J. P. L. ^{[3
,4
]}

Clevers, Hans ^{[1
,2
,3
]}

机构：

[1] Royal Netherlands Acad Arts & Sci KNAW, Hubrecht Inst, NL-3584 CT Utrecht, Netherlands

[2] UMC Utrecht, NL-3584 CT Utrecht, Netherlands

[3] UMC Utrecht, Canc Genom Netherlands, NL-3584 CG Utrecht, Netherlands

[4] UMC Utrecht, Ctr Mol Med, Mol Canc Res, NL-3584 CG Utrecht, Netherlands

[5] AMC, Ctr Expt Mol Med, Lab Expt Oncol & Radiobiol, NL-1105 AZ Amsterdam, Netherlands

[6] UMC Utrecht, Dept Med Genet, NL-3508 AB Utrecht, Netherlands

[7] UMC Utrecht, Dept Pathol, NL-3584 CX Utrecht, Netherlands

[8] Fdn Hubrecht Organoid Technol HUB, NL-3584 CT Utrecht, Netherlands

来源：

NATURE | 2015年 / 521卷 / 7550期

关键词：

IN-VITRO EXPANSION; STRUCTURAL BASIS; ORGANOIDS; ADENOMAS; REVEALS; COLON;

D O I：

10.1038/nature14415

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：43 / U329

页数：23

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