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A haploid genetic screen identifies the G1/S regulatory machinery as a determinant of Wee1 inhibitor sensitivity

Heijink, Anne Margriet ; Blomen, Vincent A. ; Bisteau, Xavier ; Degener, Fabian ; Matsushita, Felipe Yu ; Kaldis, Philipp LU ; Foijer, Floris and Van Vugt, Marcel A.T.M. (2015) In Proceedings of the National Academy of Sciences of the United States of America 112(49). p.15160-15165
Abstract

The Wee1 cell cycle checkpoint kinase prevents premature mitotic entry by inhibiting cyclin-dependent kinases. Chemical inhibitors of Wee1 are currently being tested clinically as targeted anticancer drugs. Wee1 inhibition is thought to be preferentially cytotoxic in p53-defective cancer cells. However, TP53 mutant cancers do not respond consistently to Wee1 inhibitor treatment, indicating the existence of genetic determinants of Wee1 inhibitor sensitivity other than TP53 status. To optimally facilitate patient selection for Wee1 inhibition and uncover potential resistance mechanisms, identification of these currently unknown genes is necessary. The aim of this study was therefore to identify gene mutations that determine Wee1 inhibitor... (More)

The Wee1 cell cycle checkpoint kinase prevents premature mitotic entry by inhibiting cyclin-dependent kinases. Chemical inhibitors of Wee1 are currently being tested clinically as targeted anticancer drugs. Wee1 inhibition is thought to be preferentially cytotoxic in p53-defective cancer cells. However, TP53 mutant cancers do not respond consistently to Wee1 inhibitor treatment, indicating the existence of genetic determinants of Wee1 inhibitor sensitivity other than TP53 status. To optimally facilitate patient selection for Wee1 inhibition and uncover potential resistance mechanisms, identification of these currently unknown genes is necessary. The aim of this study was therefore to identify gene mutations that determine Wee1 inhibitor sensitivity. We performed a genome-wide unbiased functional genetic screen in TP53 mutant near-haploid KBM-7 cells using gene-trap insertional mutagenesis. Insertion site mapping of cells that survived long-term Wee1 inhibition revealed enrichment of G1/S regulatory genes, including SKP2, CUL1, and CDK2. Stable depletion of SKP2, CUL1, or CDK2 or chemical Cdk2 inhibition rescued the γ-H2AX induction and abrogation of G2 phase as induced by Wee1 inhibition in breast and ovarian cancer cell lines. Remarkably, live cell imaging showed that depletion of SKP2, CUL1, or CDK2 did not rescue the Wee1 inhibition-induced karyokinesis and cytokinesis defects. These data indicate that the activity of the DNA replication machinery, beyond TP53 mutation status, determines Wee1 inhibitor sensitivity, and could serve as a selection criterion for Wee1-inhibitor eligible patients. Conversely, loss of the identified S-phase genes could serve as a mechanism of acquired resistance, which goes along with development of severe genomic instability.

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author
publishing date
type
Contribution to journal
publication status
published
subject
keywords
AZD-1775, Cell cycle, Checkpoint, MK-1775, Polyploidy
in
Proceedings of the National Academy of Sciences of the United States of America
volume
112
issue
49
pages
15160 - 15165
publisher
National Acad Sciences
external identifiers
  • pmid:26598692
  • scopus:84949466942
ISSN
0027-8424
DOI
10.1073/pnas.1505283112
language
English
LU publication?
no
id
42cfba3c-8cde-4eec-98b6-8acb845e9c9d
date added to LUP
2019-09-18 13:49:26
date last changed
2020-01-30 03:54:40
@article{42cfba3c-8cde-4eec-98b6-8acb845e9c9d,
  abstract     = {<p>The Wee1 cell cycle checkpoint kinase prevents premature mitotic entry by inhibiting cyclin-dependent kinases. Chemical inhibitors of Wee1 are currently being tested clinically as targeted anticancer drugs. Wee1 inhibition is thought to be preferentially cytotoxic in p53-defective cancer cells. However, TP53 mutant cancers do not respond consistently to Wee1 inhibitor treatment, indicating the existence of genetic determinants of Wee1 inhibitor sensitivity other than TP53 status. To optimally facilitate patient selection for Wee1 inhibition and uncover potential resistance mechanisms, identification of these currently unknown genes is necessary. The aim of this study was therefore to identify gene mutations that determine Wee1 inhibitor sensitivity. We performed a genome-wide unbiased functional genetic screen in TP53 mutant near-haploid KBM-7 cells using gene-trap insertional mutagenesis. Insertion site mapping of cells that survived long-term Wee1 inhibition revealed enrichment of G<sub>1</sub>/S regulatory genes, including SKP2, CUL1, and CDK2. Stable depletion of SKP2, CUL1, or CDK2 or chemical Cdk2 inhibition rescued the γ-H2AX induction and abrogation of G<sub>2</sub> phase as induced by Wee1 inhibition in breast and ovarian cancer cell lines. Remarkably, live cell imaging showed that depletion of SKP2, CUL1, or CDK2 did not rescue the Wee1 inhibition-induced karyokinesis and cytokinesis defects. These data indicate that the activity of the DNA replication machinery, beyond TP53 mutation status, determines Wee1 inhibitor sensitivity, and could serve as a selection criterion for Wee1-inhibitor eligible patients. Conversely, loss of the identified S-phase genes could serve as a mechanism of acquired resistance, which goes along with development of severe genomic instability.</p>},
  author       = {Heijink, Anne Margriet and Blomen, Vincent A. and Bisteau, Xavier and Degener, Fabian and Matsushita, Felipe Yu and Kaldis, Philipp and Foijer, Floris and Van Vugt, Marcel A.T.M.},
  issn         = {0027-8424},
  language     = {eng},
  month        = {12},
  number       = {49},
  pages        = {15160--15165},
  publisher    = {National Acad Sciences},
  series       = {Proceedings of the National Academy of Sciences of the United States of America},
  title        = {A haploid genetic screen identifies the G<sub>1</sub>/S regulatory machinery as a determinant of Wee1 inhibitor sensitivity},
  url          = {http://dx.doi.org/10.1073/pnas.1505283112},
  doi          = {10.1073/pnas.1505283112},
  volume       = {112},
  year         = {2015},
}