A genome-wide enrichment screen identifies NUMA1-loss as a resistance mechanism against mitotic cell-death induced by BMI1 inhibition

PLoS One. 2020 Apr 28;15(4):e0227592. doi: 10.1371/journal.pone.0227592. eCollection 2020.

Abstract

BMI1 is a core protein of the polycomb repressive complex 1 (PRC1) that is overexpressed in several cancer types, making it a promising target for cancer therapies. However, the underlying mechanisms and interactions associated with BMI1-induced tumorigenesis are often context-dependent and complex. Here, we performed a drug resistance screen on mutagenized human haploid HAP1 cells treated with BMI1 inhibitor PTC-318 to find new genetic and mechanistic features associated with BMI1-dependent cancer cell proliferation. Our screen identified NUMA1-mutations as the most significant inducer of PTC-318 cell death resistance. Independent validations on NUMA1-proficient HAP1 and non-small cell lung cancer cell lines exposed to BMI1 inhibition by PTC-318 or BMI1 knockdown resulted in cell death following mitotic arrest. Interestingly, cells with CRISPR-Cas9 derived NUMA1 knockout also showed a mitotic arrest phenotype following BMI1 inhibition but, contrary to cells with wildtype NUMA1, these cells were resistant to BMI1-dependent cell death. The current study brings new insights to BMI1 inhibition-induced mitotic lethality in cancer cells and presents a previously unknown role of NUMA1 in this process.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Antineoplastic Agents / therapeutic use
  • Apoptosis / drug effects
  • Apoptosis / genetics
  • CRISPR-Cas Systems / genetics
  • Carcinogenesis / drug effects
  • Carcinogenesis / genetics*
  • Cell Cycle Proteins / genetics*
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cell Proliferation / genetics
  • Drug Resistance, Neoplasm / genetics*
  • Gene Expression Regulation, Neoplastic
  • Gene Knockdown Techniques
  • Gene Knockout Techniques
  • Humans
  • M Phase Cell Cycle Checkpoints / drug effects
  • Mutation
  • Neoplasms / drug therapy
  • Neoplasms / genetics*
  • Neoplasms / pathology
  • Polycomb Repressive Complex 1 / antagonists & inhibitors
  • Polycomb Repressive Complex 1 / genetics
  • Polycomb Repressive Complex 1 / metabolism*
  • RNA, Small Interfering / metabolism

Substances

  • Antineoplastic Agents
  • BMI1 protein, human
  • Cell Cycle Proteins
  • NUMA1 protein, human
  • RNA, Small Interfering
  • Polycomb Repressive Complex 1

Grants and funding

MVL and SGB: Netherlands Institute of regenerative medicine (NIRM) 58473. MVL and SGB: Netherlands organization for scientific research (NWO) 823.02.007. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.