CRISPR/Cas9-derived models of ovarian high grade serous carcinoma targeting Brca1, Pten and Nf1, and correlation with platinum sensitivity

Sci Rep. 2017 Dec 4;7(1):16827. doi: 10.1038/s41598-017-17119-1.

Abstract

Transplantable murine models of ovarian high grade serous carcinoma (HGSC) remain an important research tool. We previously showed that ID8, a widely-used syngeneic model of ovarian cancer, lacked any of the frequent mutations in HGSC, and used CRISPR/Cas9 gene editing to generate derivatives with deletions in Trp53 and Brca2. Here we have used one ID8 Trp53 -/- clone to generate further mutants, with additional mutations in Brca1, Pten and Nf1, all of which are frequently mutated or deleted in HGSC. We have also generated clones with triple deletions in Trp53, Brca2 and Pten. We show that ID8 Trp53 -/-;Brca1 -/- and Trp53 -/-;Brca2 -/- cells have defective homologous recombination and increased sensitivity to both platinum and PARP inhibitor chemotherapy compared to Trp53 -/-. By contrast, loss of Pten or Nf1 increases growth rate in vivo, and reduces survival following cisplatin chemotherapy in vivo. Finally, we have also targeted Trp53 in cells isolated from a previous transgenic murine fallopian tube carcinoma model, and confirmed that loss of p53 expression in this second model accelerates intraperitoneal growth. Together, these CRISPR-generated models represent a new and simple tool to investigate the biology of HGSC, and the ID8 cell lines are freely available to researchers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • BRCA1 Protein
  • BRCA2 Protein / deficiency
  • BRCA2 Protein / genetics
  • CRISPR-Cas Systems / genetics*
  • Cell Line, Tumor
  • Cell Survival
  • Cystadenocarcinoma, Serous / drug therapy*
  • Cystadenocarcinoma, Serous / genetics
  • Cystadenocarcinoma, Serous / mortality
  • Cystadenocarcinoma, Serous / pathology
  • Disease Models, Animal
  • Female
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Neoplasm Grading
  • Neurofibromin 1 / genetics*
  • Ovarian Neoplasms / drug therapy*
  • Ovarian Neoplasms / genetics
  • Ovarian Neoplasms / mortality
  • Ovarian Neoplasms / pathology
  • PTEN Phosphohydrolase / deficiency
  • PTEN Phosphohydrolase / genetics*
  • Platinum / chemistry
  • Platinum / pharmacology
  • Platinum / therapeutic use*
  • Poly(ADP-ribose) Polymerase Inhibitors / therapeutic use
  • Survival Rate
  • Tumor Suppressor Protein p53 / deficiency
  • Tumor Suppressor Protein p53 / genetics
  • Tumor Suppressor Proteins / deficiency
  • Tumor Suppressor Proteins / genetics*

Substances

  • BRCA1 Protein
  • BRCA2 Protein
  • Brca1 protein, mouse
  • Neurofibromin 1
  • Poly(ADP-ribose) Polymerase Inhibitors
  • Tumor Suppressor Protein p53
  • Tumor Suppressor Proteins
  • Platinum
  • PTEN Phosphohydrolase
  • Pten protein, mouse