Recapitulating the adenoma-carcinoma sequence by selection of four spontaneous oncogenic mutations in mismatch-repair-deficient human colon organoids

Tomohiro Mizutani, Matteo Boretto, Sangho Lim, Jarno Drost, Diego Montiel González, Rurika Oka, Maarten H Geurts, Harry Begthel, Jeroen Korving, Johan H van Es, Ruben van Boxtel, Hans Clevers

Research output: Contribution to journalArticlepeer-review

Abstract

Carcinogenesis results from the sequential acquisition of oncogenic mutations that convert normal cells into invasive, metastasizing cancer cells. Colorectal cancer exemplifies this process through its well-described adenoma-carcinoma sequence, modeled previously using clustered regularly interspaced short palindromic repeats (CRISPR) to induce four consecutive mutations in wild-type human gut organoids. Here, we demonstrate that long-term culture of mismatch-repair-deficient organoids allows the selection of spontaneous oncogenic mutations through the sequential withdrawal of Wnt agonists, epidermal growth factor (EGF) agonists and the bone morphogenetic protein (BMP) antagonist Noggin, while TP53 mutations were selected through the addition of Nutlin-3. Thus, organoids sequentially acquired mutations in AXIN1 and AXIN2 (Wnt pathway), TP53, ACVR2A and BMPR2 (BMP pathway) and NRAS (EGF pathway), gaining complete independence from stem cell niche factors. Quadruple-pathway (Wnt, EGF receptor, p53 and BMP) mutant organoids formed solid tumors upon xenotransplantation. This demonstrates that carcinogenesis can be recapitulated in a DNA repair-mutant background through in vitro selection that targets four consecutive cancer pathways.

Original languageEnglish
Pages (from-to)1852-1867
Number of pages16
JournalNature Cancer
Volume5
Issue number12
Early online date1 Nov 2024
DOIs
Publication statusPublished - Dec 2024

Keywords

  • Adenoma/genetics
  • Animals
  • Carcinogenesis/genetics
  • Colon/pathology
  • Colonic Neoplasms/genetics
  • DNA Mismatch Repair
  • Humans
  • Mice
  • Mutation
  • Organoids/pathology
  • Tumor Suppressor Protein p53/genetics
  • Wnt Signaling Pathway/genetics

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