Sequential cancer mutations in cultured human intestinal stem cells.

Nature 2015 May 29;521(7550):43-7. Epub 2015 Apr 29.

1] Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and UMC Utrecht, 3584CT Utrecht, The Netherlands [2] Cancer Genomics Netherlands, UMC Utrecht, 3584CG Utrecht, The Netherlands.

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.

Download full-text PDF

Source
http://dx.doi.org/10.1038/nature14415DOI Listing
May 2015
124 Reads

Publication Analysis

Top Keywords

stem cells
16
intestinal stem
12
human intestinal
12
quadruple mutants
8
cultured human
8
stem-cell-niche factors
8
mutants grow
8
apc p53
8
cells
5
mutant organoids
4
smad4 cultured
4
kras smad4
4
organoids selected
4
cells mutant
4
factors culture
4
growth factors
4
individual growth
4
removing individual
4
tp53 kras
4
selected removing
4

References

(Supplied by CrossRef)

ER Fearon et al.
Cell 1990

ER Fearon et al.
Annu. Rev. Pathol. 2011

N Barker et al.
Nature 2009

AG Schepers et al.
Science 2012

L Zhu et al.
Nature 2009

X Li et al.
Nature Med. 2014

T Sato et al.
Nature 2009

T Sato et al.
Gastroenterology 2011

T Sato et al.
Methods Mol. Biol. 2013

P Jung et al.
Nature Med. 2011

Similar Publications