A platform of functional studies of ESCC-associated gene mutations identifies the roles of TGFBR2 in ESCC progression and metastasis

Cell Rep. 2024 Nov 26;43(11):114952. doi: 10.1016/j.celrep.2024.114952. Epub 2024 Nov 10.

Abstract

Genomics studies have detected numerous genetic alterations in esophageal squamous cell carcinoma (ESCC). However, the functions of these mutations largely remain elusive, partially due to a lack of feasible animal models. Here, we report a convenient platform with CRISPR-Cas9-mediated introduction of genetic alterations and orthotopic transplantation to generate a series of primary ESCC models in mice. With this platform, we validate multiple frequently mutated genes, including EP300, FAT1/2/4, KMT2D, NOTCH2, and TGFBR2, as tumor-suppressor genes in ESCC. Among them, TGFBR2 loss dramatically promotes tumorigenesis and multi-organ metastasis. Paradoxically, TGFBR2 deficiency leads to Smad3 activation, and disruption of Smad3 partially restrains the progression of Tgfbr2-mutated tumors. Drug screening with tumor organoids identifies that pinaverium bromide represses Smad3 activity and restrains Tgfbr2-deficient ESCC. Our studies provide a highly efficient platform to investigate the in vivo functions of ESCC-associated mutations and develop potential treatments for this miserable malignancy.

Keywords: CP: Cancer; TGFBR2; esophageal squamous cell carcinoma; mouse model; precise treatment.

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics
  • Cell Line, Tumor
  • Disease Progression*
  • Esophageal Neoplasms / genetics
  • Esophageal Neoplasms / metabolism
  • Esophageal Neoplasms / pathology
  • Esophageal Squamous Cell Carcinoma* / genetics
  • Esophageal Squamous Cell Carcinoma* / metabolism
  • Esophageal Squamous Cell Carcinoma* / pathology
  • Humans
  • Mice
  • Mutation* / genetics
  • Neoplasm Metastasis*
  • Receptor, Transforming Growth Factor-beta Type II* / genetics
  • Receptor, Transforming Growth Factor-beta Type II* / metabolism
  • Smad3 Protein* / genetics
  • Smad3 Protein* / metabolism

Substances

  • Receptor, Transforming Growth Factor-beta Type II
  • Smad3 Protein
  • TGFBR2 protein, human