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

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.
Graham MacLeod 1 Danielle A Bozek 2 Nishani Rajakulendran 1 Vernon Monteiro 1 Moloud Ahmadi 1 Zachary Steinhart 1 Michelle M Kushida 3 Helen Yu 3 Fiona J Coutinho 3 Florence M G Cavalli 3 Ian Restall 2 Xiaoguang Hao 2 Traver Hart 4 H Artee Luchman 2 Samuel Weiss 2 Peter B Dirks 5 Stephane Angers 6
Affiliations

Affiliations

  • 1 Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, Canada.
  • 2 Hotchkiss Brain Institute, Department of Cell Biology and Anatomy, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada.
  • 3 Developmental and Stem Cell Biology Program and Arthur and Sonia Labatt Brain Tumour Research Centre, The Hospital for Sick Children, Toronto, ON, Canada.
  • 4 Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.
  • 5 Developmental and Stem Cell Biology Program and Arthur and Sonia Labatt Brain Tumour Research Centre, The Hospital for Sick Children, Toronto, ON, Canada; Department of Molecular Genetics, Department of Laboratory Medicine and Pathobiology, Division of Neurosurgery, Department of Surgery, Faculty of Medicine, University of Toronto, Toronto, ON, Canada; Division of Neurosurgery, The Hospital for Sick Children, Toronto, ON, Canada. Electronic address: peter.dirks@sickkids.ca.
  • 6 Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, Canada; Department of Biochemistry, Faculty of Medicine, University of Toronto, Toronto, ON, Canada. Electronic address: stephane.angers@utoronto.ca.
Abstract

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.

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