Cargando…

Targeting the NPL4 Adaptor of p97/VCP Segregase by Disulfiram as an Emerging Cancer Vulnerability Evokes Replication Stress and DNA Damage while Silencing the ATR Pathway

Research on repurposing the old alcohol-aversion drug disulfiram (DSF) for cancer treatment has identified inhibition of NPL4, an adaptor of the p97/VCP segregase essential for turnover of proteins involved in multiple pathways, as an unsuspected cancer cell vulnerability. While we reported that NPL...

Descripción completa

Detalles Bibliográficos
Autores principales: Majera, Dusana, Skrott, Zdenek, Chroma, Katarina, Merchut-Maya, Joanna Maria, Mistrik, Martin, Bartek, Jiri
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7072750/
https://www.ncbi.nlm.nih.gov/pubmed/32085572
http://dx.doi.org/10.3390/cells9020469
_version_ 1783506479323545600
author Majera, Dusana
Skrott, Zdenek
Chroma, Katarina
Merchut-Maya, Joanna Maria
Mistrik, Martin
Bartek, Jiri
author_facet Majera, Dusana
Skrott, Zdenek
Chroma, Katarina
Merchut-Maya, Joanna Maria
Mistrik, Martin
Bartek, Jiri
author_sort Majera, Dusana
collection PubMed
description Research on repurposing the old alcohol-aversion drug disulfiram (DSF) for cancer treatment has identified inhibition of NPL4, an adaptor of the p97/VCP segregase essential for turnover of proteins involved in multiple pathways, as an unsuspected cancer cell vulnerability. While we reported that NPL4 is targeted by the anticancer metabolite of DSF, the bis-diethyldithiocarbamate-copper complex (CuET), the exact, apparently multifaceted mechanism(s) through which the CuET-induced aggregation of NPL4 kills cancer cells remains to be fully elucidated. Given the pronounced sensitivity to CuET in tumor cell lines lacking the genome integrity caretaker proteins BRCA1 and BRCA2, here we investigated the impact of NPL4 targeting by CuET on DNA replication dynamics and DNA damage response pathways in human cancer cell models. Our results show that CuET treatment interferes with DNA replication, slows down replication fork progression and causes accumulation of single-stranded DNA (ssDNA). Such a replication stress (RS) scenario is associated with DNA damage, preferentially in the S phase, and activates the homologous recombination (HR) DNA repair pathway. At the same time, we find that cellular responses to the CuET-triggered RS are seriously impaired due to concomitant malfunction of the ATRIP-ATR-CHK1 signaling pathway that reflects an unorthodox checkpoint silencing mode through ATR (Ataxia telangiectasia and Rad3 related) kinase sequestration within the CuET-evoked NPL4 protein aggregates.
format Online
Article
Text
id pubmed-7072750
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-70727502020-03-19 Targeting the NPL4 Adaptor of p97/VCP Segregase by Disulfiram as an Emerging Cancer Vulnerability Evokes Replication Stress and DNA Damage while Silencing the ATR Pathway Majera, Dusana Skrott, Zdenek Chroma, Katarina Merchut-Maya, Joanna Maria Mistrik, Martin Bartek, Jiri Cells Article Research on repurposing the old alcohol-aversion drug disulfiram (DSF) for cancer treatment has identified inhibition of NPL4, an adaptor of the p97/VCP segregase essential for turnover of proteins involved in multiple pathways, as an unsuspected cancer cell vulnerability. While we reported that NPL4 is targeted by the anticancer metabolite of DSF, the bis-diethyldithiocarbamate-copper complex (CuET), the exact, apparently multifaceted mechanism(s) through which the CuET-induced aggregation of NPL4 kills cancer cells remains to be fully elucidated. Given the pronounced sensitivity to CuET in tumor cell lines lacking the genome integrity caretaker proteins BRCA1 and BRCA2, here we investigated the impact of NPL4 targeting by CuET on DNA replication dynamics and DNA damage response pathways in human cancer cell models. Our results show that CuET treatment interferes with DNA replication, slows down replication fork progression and causes accumulation of single-stranded DNA (ssDNA). Such a replication stress (RS) scenario is associated with DNA damage, preferentially in the S phase, and activates the homologous recombination (HR) DNA repair pathway. At the same time, we find that cellular responses to the CuET-triggered RS are seriously impaired due to concomitant malfunction of the ATRIP-ATR-CHK1 signaling pathway that reflects an unorthodox checkpoint silencing mode through ATR (Ataxia telangiectasia and Rad3 related) kinase sequestration within the CuET-evoked NPL4 protein aggregates. MDPI 2020-02-18 /pmc/articles/PMC7072750/ /pubmed/32085572 http://dx.doi.org/10.3390/cells9020469 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Majera, Dusana
Skrott, Zdenek
Chroma, Katarina
Merchut-Maya, Joanna Maria
Mistrik, Martin
Bartek, Jiri
Targeting the NPL4 Adaptor of p97/VCP Segregase by Disulfiram as an Emerging Cancer Vulnerability Evokes Replication Stress and DNA Damage while Silencing the ATR Pathway
title Targeting the NPL4 Adaptor of p97/VCP Segregase by Disulfiram as an Emerging Cancer Vulnerability Evokes Replication Stress and DNA Damage while Silencing the ATR Pathway
title_full Targeting the NPL4 Adaptor of p97/VCP Segregase by Disulfiram as an Emerging Cancer Vulnerability Evokes Replication Stress and DNA Damage while Silencing the ATR Pathway
title_fullStr Targeting the NPL4 Adaptor of p97/VCP Segregase by Disulfiram as an Emerging Cancer Vulnerability Evokes Replication Stress and DNA Damage while Silencing the ATR Pathway
title_full_unstemmed Targeting the NPL4 Adaptor of p97/VCP Segregase by Disulfiram as an Emerging Cancer Vulnerability Evokes Replication Stress and DNA Damage while Silencing the ATR Pathway
title_short Targeting the NPL4 Adaptor of p97/VCP Segregase by Disulfiram as an Emerging Cancer Vulnerability Evokes Replication Stress and DNA Damage while Silencing the ATR Pathway
title_sort targeting the npl4 adaptor of p97/vcp segregase by disulfiram as an emerging cancer vulnerability evokes replication stress and dna damage while silencing the atr pathway
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7072750/
https://www.ncbi.nlm.nih.gov/pubmed/32085572
http://dx.doi.org/10.3390/cells9020469
work_keys_str_mv AT majeradusana targetingthenpl4adaptorofp97vcpsegregasebydisulfiramasanemergingcancervulnerabilityevokesreplicationstressanddnadamagewhilesilencingtheatrpathway
AT skrottzdenek targetingthenpl4adaptorofp97vcpsegregasebydisulfiramasanemergingcancervulnerabilityevokesreplicationstressanddnadamagewhilesilencingtheatrpathway
AT chromakatarina targetingthenpl4adaptorofp97vcpsegregasebydisulfiramasanemergingcancervulnerabilityevokesreplicationstressanddnadamagewhilesilencingtheatrpathway
AT merchutmayajoannamaria targetingthenpl4adaptorofp97vcpsegregasebydisulfiramasanemergingcancervulnerabilityevokesreplicationstressanddnadamagewhilesilencingtheatrpathway
AT mistrikmartin targetingthenpl4adaptorofp97vcpsegregasebydisulfiramasanemergingcancervulnerabilityevokesreplicationstressanddnadamagewhilesilencingtheatrpathway
AT bartekjiri targetingthenpl4adaptorofp97vcpsegregasebydisulfiramasanemergingcancervulnerabilityevokesreplicationstressanddnadamagewhilesilencingtheatrpathway