A kinome screen identifies checkpoint kinase 1 (CHK1) as a sensitizer for RRM1-dependent gemcitabine efficacy.

Gemcitabine is among the most efficacious and widely used antimetabolite agents. Its molecular targets are ribonucleotide reductase M1 (RRM1) and elongating DNA. Acquired and de novo resistance as a result of RRM1 overexpression are major obstacles to therapeutic efficacy. We deployed a synthetic le...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Jun Zhou, Zhengming Chen, Agnes Malysa, Xin Li, Paula Oliveira, Yingtao Zhang, Gerold Bepler
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1f6fb0eb1d5d46e69fff77bbf52c2a35
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:1f6fb0eb1d5d46e69fff77bbf52c2a35
record_format	dspace
spelling	oai:doaj.org-article:1f6fb0eb1d5d46e69fff77bbf52c2a352021-11-18T07:55:01ZA kinome screen identifies checkpoint kinase 1 (CHK1) as a sensitizer for RRM1-dependent gemcitabine efficacy.1932-620310.1371/journal.pone.0058091https://doaj.org/article/1f6fb0eb1d5d46e69fff77bbf52c2a352013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23483975/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Gemcitabine is among the most efficacious and widely used antimetabolite agents. Its molecular targets are ribonucleotide reductase M1 (RRM1) and elongating DNA. Acquired and de novo resistance as a result of RRM1 overexpression are major obstacles to therapeutic efficacy. We deployed a synthetic lethality screen to investigate if knockdown of 87 selected protein kinases by siRNA could overcome RRM1-dependent gemcitabine resistance in high and low RRM1-expressing model systems. The models included genetically RRM1-modified lung and breast cancer cell lines, cell lines with gemcitabine-induced RRM1 overexpression, and a series of naturally gemcitabine-resistant cell lines. Lead molecular targets were validated by determination of differential gemcitabine activity using cell lines with and without target knock down, and by assessing synergistic activity between gemcitabine and an inhibitor of the lead target. CHK1 was identified has the kinase with the most significant and robust interaction, and it was validated using AZD7762, a small-molecule ATP-competitive inhibitor of CHK1 activation. Synergism between CHK1 inhibition and RRM1-dependent gemcitabine efficacy was observed in cells with high RRM1 levels, while antagonism was observed in cells with low RRM1 levels. In addition, four cell lines with natural gemcitabine resistance demonstrated improved gemcitabine efficacy after CHK1 inhibition. In tumor specimens from 187 patients with non-small-cell lung cancer, total CHK1 and RRM1 in situ protein levels were significantly (p = 0.003) and inversely correlated. We conclude that inhibition of CHK1 may have its greatest clinical utility in malignancies where gemcitabine resistance is a result of elevated RRM1 levels. We also conclude that CHK1 inhibition in tumors with low RRM1 levels may be detrimental to gemcitabine efficacy.Jun ZhouZhengming ChenAgnes MalysaXin LiPaula OliveiraYingtao ZhangGerold BeplerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 3, p e58091 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jun Zhou Zhengming Chen Agnes Malysa Xin Li Paula Oliveira Yingtao Zhang Gerold Bepler A kinome screen identifies checkpoint kinase 1 (CHK1) as a sensitizer for RRM1-dependent gemcitabine efficacy.
description	Gemcitabine is among the most efficacious and widely used antimetabolite agents. Its molecular targets are ribonucleotide reductase M1 (RRM1) and elongating DNA. Acquired and de novo resistance as a result of RRM1 overexpression are major obstacles to therapeutic efficacy. We deployed a synthetic lethality screen to investigate if knockdown of 87 selected protein kinases by siRNA could overcome RRM1-dependent gemcitabine resistance in high and low RRM1-expressing model systems. The models included genetically RRM1-modified lung and breast cancer cell lines, cell lines with gemcitabine-induced RRM1 overexpression, and a series of naturally gemcitabine-resistant cell lines. Lead molecular targets were validated by determination of differential gemcitabine activity using cell lines with and without target knock down, and by assessing synergistic activity between gemcitabine and an inhibitor of the lead target. CHK1 was identified has the kinase with the most significant and robust interaction, and it was validated using AZD7762, a small-molecule ATP-competitive inhibitor of CHK1 activation. Synergism between CHK1 inhibition and RRM1-dependent gemcitabine efficacy was observed in cells with high RRM1 levels, while antagonism was observed in cells with low RRM1 levels. In addition, four cell lines with natural gemcitabine resistance demonstrated improved gemcitabine efficacy after CHK1 inhibition. In tumor specimens from 187 patients with non-small-cell lung cancer, total CHK1 and RRM1 in situ protein levels were significantly (p = 0.003) and inversely correlated. We conclude that inhibition of CHK1 may have its greatest clinical utility in malignancies where gemcitabine resistance is a result of elevated RRM1 levels. We also conclude that CHK1 inhibition in tumors with low RRM1 levels may be detrimental to gemcitabine efficacy.
format	article
author	Jun Zhou Zhengming Chen Agnes Malysa Xin Li Paula Oliveira Yingtao Zhang Gerold Bepler
author_facet	Jun Zhou Zhengming Chen Agnes Malysa Xin Li Paula Oliveira Yingtao Zhang Gerold Bepler
author_sort	Jun Zhou
title	A kinome screen identifies checkpoint kinase 1 (CHK1) as a sensitizer for RRM1-dependent gemcitabine efficacy.
title_short	A kinome screen identifies checkpoint kinase 1 (CHK1) as a sensitizer for RRM1-dependent gemcitabine efficacy.
title_full	A kinome screen identifies checkpoint kinase 1 (CHK1) as a sensitizer for RRM1-dependent gemcitabine efficacy.
title_fullStr	A kinome screen identifies checkpoint kinase 1 (CHK1) as a sensitizer for RRM1-dependent gemcitabine efficacy.
title_full_unstemmed	A kinome screen identifies checkpoint kinase 1 (CHK1) as a sensitizer for RRM1-dependent gemcitabine efficacy.
title_sort	kinome screen identifies checkpoint kinase 1 (chk1) as a sensitizer for rrm1-dependent gemcitabine efficacy.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/1f6fb0eb1d5d46e69fff77bbf52c2a35
work_keys_str_mv	AT junzhou akinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy AT zhengmingchen akinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy AT agnesmalysa akinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy AT xinli akinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy AT paulaoliveira akinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy AT yingtaozhang akinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy AT geroldbepler akinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy AT junzhou kinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy AT zhengmingchen kinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy AT agnesmalysa kinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy AT xinli kinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy AT paulaoliveira kinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy AT yingtaozhang kinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy AT geroldbepler kinomescreenidentifiescheckpointkinase1chk1asasensitizerforrrm1dependentgemcitabineefficacy
_version_	1718422716683386880

A kinome screen identifies checkpoint kinase 1 (CHK1) as a sensitizer for RRM1-dependent gemcitabine efficacy.

Ejemplares similares