Genome-Wide CRISPR-Cas9 Screens Expose Genetic Vulnerabilities and Mechanisms of Temozolomide Sensitivity in Glioblastoma Stem Cells

Cell Rep. 2019 Apr 16;27(3):971-986.e9. doi: 10.1016/j.celrep.2019.03.047.


Glioblastoma therapies have remained elusive due to limitations in understanding mechanisms of growth and survival of the tumorigenic population. Using CRISPR-Cas9 approaches in patient-derived GBM stem cells (GSCs) to interrogate function of the coding genome, we identify actionable pathways responsible for growth, which reveal the gene-essential circuitry of GBM stemness and proliferation. In particular, we characterize members of the SOX transcription factor family, SOCS3, USP8, and DOT1L, and protein ufmylation as important for GSC growth. Additionally, we reveal mechanisms of temozolomide resistance that could lead to combination strategies. By reaching beyond static genome analysis of bulk tumors, with a genome-wide functional approach, we reveal genetic dependencies within a broad range of biological processes to provide increased understanding of GBM growth and treatment resistance.

Keywords: CRISPR-Cas9; fitness genes; functional genomics; glioblastoma; glioblastoma stem cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Brain Neoplasms / drug therapy
  • Brain Neoplasms / mortality
  • Brain Neoplasms / pathology*
  • CRISPR-Cas Systems / genetics*
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Drug Resistance, Neoplasm / genetics
  • Endopeptidases / genetics
  • Endopeptidases / metabolism
  • Endosomal Sorting Complexes Required for Transport / genetics
  • Endosomal Sorting Complexes Required for Transport / metabolism
  • Female
  • Gene Editing / methods*
  • Gene Expression Regulation, Neoplastic / drug effects
  • Gene Library
  • Glioblastoma / drug therapy
  • Glioblastoma / mortality
  • Glioblastoma / pathology*
  • Histone Methyltransferases / metabolism
  • Humans
  • Mice
  • Mice, SCID
  • Neoplastic Stem Cells / drug effects
  • Neoplastic Stem Cells / metabolism*
  • Suppressor of Cytokine Signaling 3 Protein / genetics
  • Suppressor of Cytokine Signaling 3 Protein / metabolism
  • Survival Analysis
  • Temozolomide / pharmacology*
  • Temozolomide / therapeutic use
  • Ubiquitin Thiolesterase / genetics
  • Ubiquitin Thiolesterase / metabolism


  • Endosomal Sorting Complexes Required for Transport
  • SOCS3 protein, human
  • Suppressor of Cytokine Signaling 3 Protein
  • Histone Methyltransferases
  • Endopeptidases
  • USP8 protein, human
  • Ubiquitin Thiolesterase
  • Temozolomide