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

Research output: Contribution to journalArticle

344 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)256-262
Number of pages7
JournalNature Medicine
Volume21
Issue number3
DOIs
Publication statusPublished - 2015

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Clustered Regularly Interspaced Short Palindromic Repeats
Organoids
Human Engineering
Tumors
Colorectal Neoplasms
Mutation
Stem cells
Genes
Cell signaling
Gene encoding
Stem Cell Niche
Chromosomes
Neoplasm Micrometastasis
Phosphatidylinositol 3-Kinases
Tumor Microenvironment
Neoplastic Stem Cells
Liver
Human Chromosomes
Intestinal Mucosa
Tumor Suppressor Genes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Modeling colorectal cancer using CRISPR-Cas9-mediated engineering of human intestinal organoids. / Matano, Mami; Date, Shoichi; Shimokawa, Mariko; Takano, Ai; Fujii, Masayuki; Ohta, Yuki; Watanabe, Toshiaki; Kanai, Takanori; Sato, Toshiro.

In: Nature Medicine, Vol. 21, No. 3, 2015, p. 256-262.

Research output: Contribution to journalArticle

Matano, M, Date, S, Shimokawa, M, Takano, A, Fujii, M, Ohta, Y, Watanabe, T, Kanai, T & Sato, T 2015, 'Modeling colorectal cancer using CRISPR-Cas9-mediated engineering of human intestinal organoids', Nature Medicine, vol. 21, no. 3, pp. 256-262. https://doi.org/10.1038/nm.3802
Matano, Mami ; Date, Shoichi ; Shimokawa, Mariko ; Takano, Ai ; Fujii, Masayuki ; Ohta, Yuki ; Watanabe, Toshiaki ; Kanai, Takanori ; Sato, Toshiro. / Modeling colorectal cancer using CRISPR-Cas9-mediated engineering of human intestinal organoids. In: Nature Medicine. 2015 ; Vol. 21, No. 3. pp. 256-262.
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