Modeling colorectal cancer using CRISPR-Cas9-mediated engineering of human intestinal organoids

Mami Matano, Shoichi Date, Mariko Shimokawa, Ai Takano, Masayuki Fujii, Yuki Ohta, Toshiaki Watanabe, Takanori Kanai, Toshiro Sato

研究成果: Article

435 被引用数 (Scopus)

抄録

Human colorectal tumors bear recurrent mutations in genes encoding proteins operative in the WNT, MAPK, TGF-β, TP53 and PI3K pathways. Although these pathways influence intestinal stem cell niche signaling, the extent to which mutations in these pathways contribute to human colorectal carcinogenesis remains unclear. Here we use the CRISPR-Cas9 genome-editing system to introduce multiple such mutations into organoids derived from normal human intestinal epithelium. By modulating the culture conditions to mimic that of the intestinal niche, we selected isogenic organoids harboring mutations in the tumor suppressor genes APC, SMAD4 and TP53, and in the oncogenes KRAS and/or PIK3CA. Organoids engineered to express all five mutations grew independently of niche factors in vitro, and they formed tumors after implantation under the kidney subcapsule in mice. Although they formed micrometastases containing dormant tumor-initiating cells after injection into the spleen of mice, they failed to colonize in the liver. In contrast, engineered organoids derived from chromosome-instable human adenomas formed macrometastatic colonies. These results suggest that 'driver' pathway mutations enable stem cell maintenance in the hostile tumor microenvironment, but that additional molecular lesions are required for invasive behavior.

本文言語English
ページ(範囲)256-262
ページ数7
ジャーナルNature medicine
21
3
DOI
出版ステータスPublished - 2015 3

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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