Haploid genetic screens identify genetic vulnerabilities to microtubule-targeting agents

Mol Oncol. 2018 Jun;12(6):953-971. doi: 10.1002/1878-0261.12307. Epub 2018 May 1.

Abstract

The absence of biomarkers to accurately predict anticancer therapy response remains a major obstacle in clinical oncology. We applied a genome-wide loss-of-function screening approach in human haploid cells to characterize genetic vulnerabilities to classical microtubule-targeting agents. Using docetaxel and vinorelbine, two well-established chemotherapeutic agents, we sought to identify genetic alterations sensitizing human HAP1 cells to these drugs. Despite the fact that both drugs act on microtubules, a set of distinct genes were identified whose disruption affects drug sensitivity. For docetaxel, this included a number of genes with a function in mitosis, while for vinorelbine we identified inactivation of FBXW7, RB1, and NF2, three frequently mutated tumor suppressor genes, as sensitizing factors. We validated these genes using independent knockout clones and confirmed FBXW7 as an important regulator of the mitotic spindle assembly. Upon FBXW7 depletion, vinorelbine treatment led to decreased survival of cells due to defective mitotic progression and subsequent mitotic catastrophe. We show that haploid insertional mutagenesis screens are a useful tool to study genetic vulnerabilities to classical chemotherapeutic drugs by identifying thus far unknown sensitivity factors. These results provide a rationale for investigating patient response to vinca alkaloid-based anticancer treatment in relation to the mutational status of these three tumor suppressor genes, and could in the future lead to the establishment of novel predictive biomarkers or suggest new drug combinations based on molecular mechanisms of drug sensitivity.

Keywords: FBXW7; docetaxel; haploid screens; vinorelbine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B / metabolism
  • Cell Cycle Proteins / antagonists & inhibitors
  • Cell Cycle Proteins / metabolism
  • Cell Death / drug effects
  • Cell Line, Tumor
  • F-Box-WD Repeat-Containing Protein 7 / genetics
  • Gene Expression Regulation, Neoplastic / drug effects
  • Genes, Tumor Suppressor
  • Genetic Testing*
  • Genome, Human
  • Haploidy*
  • Humans
  • Microtubules / drug effects
  • Microtubules / metabolism*
  • Mitosis / drug effects
  • Morpholines / pharmacology
  • Mutagenesis, Insertional / genetics
  • Mutation / genetics
  • Protein Serine-Threonine Kinases / antagonists & inhibitors
  • Protein Serine-Threonine Kinases / metabolism
  • Protein-Tyrosine Kinases / antagonists & inhibitors
  • Protein-Tyrosine Kinases / metabolism
  • Purines / pharmacology
  • Vinorelbine / pharmacology

Substances

  • ABCB1 protein, human
  • ATP Binding Cassette Transporter, Subfamily B
  • Cell Cycle Proteins
  • F-Box-WD Repeat-Containing Protein 7
  • FBXW7 protein, human
  • Morpholines
  • Purines
  • Protein-Tyrosine Kinases
  • Protein Serine-Threonine Kinases
  • TTK protein, human
  • Vinorelbine
  • 2-(4-morpholinoanilino)-6-cyclohexylaminopurine

Associated data

  • GENBANK/SRP058962