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 Nam; Jeongwu Lee; Steven M. Pollard; Jun Zhu; Jeffery J. Delrow; Bruce E. Clurman; James M. Olson; Patrick J. Paddison

doi:10.1016/j.celrep.2015.11.021

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

Research output: Contribution to journal › Article › peer-review

127 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 language	English
Pages (from-to)	2425-2439
Number of pages	15
Journal	Cell Reports
Volume	13
Issue number	11
DOIs	https://doi.org/10.1016/j.celrep.2015.11.021
State	Published - 22 Dec 2015

Keywords

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

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10.1016/j.celrep.2015.11.021

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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

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

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.",

keywords = "CRISPR-Cas9, Cancer therapeutics, Functional genomics, Gene editing, Glioblastoma, Myt1, PKMYT1, WEE1",

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

note = "Publisher Copyright: {\textcopyright} 2015 The Authors.",

year = "2015",

month = dec,

day = "22",

doi = "10.1016/j.celrep.2015.11.021",

language = "English",

volume = "13",

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journal = "Cell Reports",

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Toledo, CM, Ding, Y, Hoellerbauer, P, Davis, RJ, Basom, R, Girard, EJ, Lee, E, Corrin, P, Hart, T, Bolouri, H, Davison, J, Zhang, Q, Hardcastle, J, Aronow, BJ, Plaisier, CL, Baliga, NS, Moffat, J, Lin, Q, Li, XN, Nam, DH, Lee, J, Pollard, SM, Zhu, J, Delrow, JJ, Clurman, BE, Olson, JM & Paddison, PJ 2015, 'Genome-wide CRISPR-Cas9 Screens Reveal Loss of Redundancy between PKMYT1 and WEE1 in Glioblastoma Stem-like Cells', Cell Reports, vol. 13, no. 11, pp. 2425-2439. https://doi.org/10.1016/j.celrep.2015.11.021

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T1 - Genome-wide CRISPR-Cas9 Screens Reveal Loss of Redundancy between PKMYT1 and WEE1 in Glioblastoma Stem-like Cells

AU - Toledo, Chad M.

AU - Ding, Yu

AU - Hoellerbauer, Pia

AU - Davis, Ryan J.

AU - Basom, Ryan

AU - Girard, Emily J.

AU - Lee, Eunjee

AU - Corrin, Philip

AU - Hart, Traver

AU - Bolouri, Hamid

AU - Davison, Jerry

AU - Zhang, Qing

AU - Hardcastle, Justin

AU - Aronow, Bruce J.

AU - Plaisier, Christopher L.

AU - Baliga, Nitin S.

AU - Moffat, Jason

AU - Lin, Qi

AU - Li, Xiao Nan

AU - Nam, Do Hyun

AU - Lee, Jeongwu

AU - Pollard, Steven M.

AU - Zhu, Jun

AU - Delrow, Jeffery J.

AU - Clurman, Bruce E.

AU - Olson, James M.

AU - Paddison, Patrick J.

PY - 2015/12/22

Y1 - 2015/12/22

N2 - 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.

AB - 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.

KW - CRISPR-Cas9

KW - Cancer therapeutics

KW - Functional genomics

KW - Gene editing

KW - Glioblastoma

KW - Myt1

KW - PKMYT1

KW - WEE1

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DO - 10.1016/j.celrep.2015.11.021

M3 - Article

C2 - 26673326

AN - SCOPUS:84952872645

SN - 2211-1247

VL - 13

SP - 2425

EP - 2439

JO - Cell Reports

JF - Cell Reports

IS - 11

ER -

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

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Keywords

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