Genome-Wide CRISPR-Cas9 Screens Expose Genetic Vulnerabilities and Mechanisms of Temozolomide Sensitivity in Glioblastoma Stem Cells
Graham MacLeod,
Danielle A. Bozek,
Nishani Rajakulendran,
Vernon Monteiro,
Moloud Ahmadi,
Zachary Steinhart,
Michelle M. Kushida,
Helen Yu,
Fiona J. Coutinho,
Florence M.G. Cavalli,
Ian Restall,
Xiaoguang Hao,
Traver Hart,
H. Artee Luchman,
Samuel Weiss,
Peter B. Dirks,
Stephane Angers
Affiliations
Graham MacLeod
Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, Canada
Danielle A. Bozek
Hotchkiss Brain Institute, Department of Cell Biology and Anatomy, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada
Nishani Rajakulendran
Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, Canada
Vernon Monteiro
Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, Canada
Moloud Ahmadi
Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, Canada
Zachary Steinhart
Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, Canada
Michelle M. Kushida
Developmental and Stem Cell Biology Program and Arthur and Sonia Labatt Brain Tumour Research Centre, The Hospital for Sick Children, Toronto, ON, Canada
Helen Yu
Developmental and Stem Cell Biology Program and Arthur and Sonia Labatt Brain Tumour Research Centre, The Hospital for Sick Children, Toronto, ON, Canada
Fiona J. Coutinho
Developmental and Stem Cell Biology Program and Arthur and Sonia Labatt Brain Tumour Research Centre, The Hospital for Sick Children, Toronto, ON, Canada
Florence M.G. Cavalli
Developmental and Stem Cell Biology Program and Arthur and Sonia Labatt Brain Tumour Research Centre, The Hospital for Sick Children, Toronto, ON, Canada
Ian Restall
Hotchkiss Brain Institute, Department of Cell Biology and Anatomy, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada
Xiaoguang Hao
Hotchkiss Brain Institute, Department of Cell Biology and Anatomy, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada
Traver Hart
Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA
H. Artee Luchman
Hotchkiss Brain Institute, Department of Cell Biology and Anatomy, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada
Samuel Weiss
Hotchkiss Brain Institute, Department of Cell Biology and Anatomy, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada
Peter B. Dirks
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; Corresponding author
Stephane Angers
Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, Canada; Department of Biochemistry, Faculty of Medicine, University of Toronto, Toronto, ON, Canada; Corresponding author
Summary: 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. : MacLeod et al. describe genome-wide CRISPR-Cas9 screens identifying genetic vulnerabilities across a panel of patient-derived glioblastoma stem cell cultures. Regulators of stemness (SOX2, SOX9, DOT1L, and SOCS3) and stress response (ufmylation and ERAD pathways) govern the growth of glioblastoma stem cells. Chemogenomic screens using temozolomide identify modulators of sensitivity to chemotherapy. Keywords: glioblastoma, glioblastoma stem cells, CRISPR-Cas9, fitness genes, functional genomics
