Genome-wide CRISPR-Cas9 Screens Reveal Loss of Redundancy between PKMYT1 and WEE1 in Glioblastoma Stem-like Cells

Chad M. Toledo, Yu Ding, Pia Hoellerbauer, Ryan J. Davis, Ryan Basom, Emily J. Girard, Eunjee Lee, Philip Corrin, Traver Hart, Hamid Bolouri, Jerry Davison, Qing Zhang, Justin Hardcastle, Bruce J. Aronow, Christopher L. Plaisier, Nitin S. Baliga, Jason Moffat, Qi Lin, Xiao Nan Li, Do Hyun NamJeongwu Lee, Steven M. Pollard, Jun Zhu, Jeffery J. Delrow, Bruce E. Clurman, James M. Olson*, Patrick J. Paddison

*Corresponding author for this work

Research output: Contribution to journalArticle

58 Scopus citations

Abstract

To identify therapeutic targets for glioblastoma (GBM), we performed genome-wide CRISPR-Cas9 knockout (KO) screens in patient-derived GBM stem-like cells (GSCs) and human neural stem/progenitors (NSCs), non-neoplastic stem cell controls, for genes required for their in vitro growth. Surprisingly, the vast majority GSC-lethal hits were found outside of molecular networks commonly altered in GBM and GSCs (e.g., oncogenic drivers). In vitro and in vivo validation of GSC-specific targets revealed several strong hits, including the wee1-like kinase, PKMYT1/Myt1. Mechanistic studies demonstrated that PKMYT1 acts redundantly with WEE1 to inhibit cyclin B-CDK1 activity via CDK1-Y15 phosphorylation and to promote timely completion of mitosis in NSCs. However, in GSCs, this redundancy is lost, most likely as a result of oncogenic signaling, causing GBM-specific lethality.

Original languageEnglish (US)
Pages (from-to)2425-2439
Number of pages15
JournalCell reports
Volume13
Issue number11
DOIs
StatePublished - Dec 22 2015

Keywords

  • CRISPR-Cas9
  • Cancer therapeutics
  • Functional genomics
  • Gene editing
  • Glioblastoma
  • Myt1
  • PKMYT1
  • WEE1

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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    Toledo, C. M., Ding, Y., Hoellerbauer, P., Davis, R. J., Basom, R., Girard, E. J., Lee, E., Corrin, P., Hart, T., Bolouri, H., Davison, J., Zhang, Q., Hardcastle, J., Aronow, B. J., Plaisier, C. L., Baliga, N. S., Moffat, J., Lin, Q., Li, X. N., ... Paddison, P. J. (2015). Genome-wide CRISPR-Cas9 Screens Reveal Loss of Redundancy between PKMYT1 and WEE1 in Glioblastoma Stem-like Cells. Cell reports, 13(11), 2425-2439. https://doi.org/10.1016/j.celrep.2015.11.021