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Heterozygous mutations in valosin-containing protein (VCP) and resistance to VCP inhibitors

In recent years, multiple studies including ours have reported on the mechanism of resistance towards valosin-containing protein (VCP) inhibitors. While all these studies reported target alterations via mutations in VCP as the primary mechanism of resistance, discrepancies persist to date regarding...

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Autores principales: Bastola, Prabhakar, Bilkis, Rabeya, De Souza, Cristabelle, Minn, Kay, Chien, Jeremy
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6662852/
https://www.ncbi.nlm.nih.gov/pubmed/31358864
http://dx.doi.org/10.1038/s41598-019-47085-9
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author Bastola, Prabhakar
Bilkis, Rabeya
De Souza, Cristabelle
Minn, Kay
Chien, Jeremy
author_facet Bastola, Prabhakar
Bilkis, Rabeya
De Souza, Cristabelle
Minn, Kay
Chien, Jeremy
author_sort Bastola, Prabhakar
collection PubMed
description In recent years, multiple studies including ours have reported on the mechanism of resistance towards valosin-containing protein (VCP) inhibitors. While all these studies reported target alterations via mutations in VCP as the primary mechanism of resistance, discrepancies persist to date regarding the zygosity of these mutations responsible for the resistance. In addition, the extent to which resistant cells harbor additional mutations in other genes is not well described. In this study, we performed global transcript analysis of the parental and previously reported VCP inhibitor (CB-5083) resistant cells and found additional mutations in the resistant cells. However, our CRISPR-Cas9 gene editing studies indicate that specific mutations in VCP are sufficient to produce resistance to CB-5083 suggesting the importance of on-target mutations in VCP for resistance. Strikingly, our analysis indicates a preexisting heterozygous frameshift mutation at codon 616 (N616fs*) in one of the VCP alleles in HCT116 cells, and we showed that this mutant allele is subjected to the nonsense-mediated decay (NMD). Accordingly, we identified a heterozygous mutation at codon 526 (L526S) in genomic DNA sequencing but a homozygous L526S mutation in complementary DNA sequencing in our independently generated CB-5083 resistant HCT116 cells, implying that the L526S mutation occurs in the allele that does not harbor the frameshift N616fs* mutation. Our results suggest the NMD as a possible mechanism for achieving the homozygosity of VCP mutant responsible for the resistance to VCP inhibitors while resolving the discrepancies among previous studies. Our results also underscore the importance of performing simultaneous genomic and complementary DNA sequencing when attributing mutational effects on the functionality particularly for an oligomer protein like VCP.
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spelling pubmed-66628522019-08-02 Heterozygous mutations in valosin-containing protein (VCP) and resistance to VCP inhibitors Bastola, Prabhakar Bilkis, Rabeya De Souza, Cristabelle Minn, Kay Chien, Jeremy Sci Rep Article In recent years, multiple studies including ours have reported on the mechanism of resistance towards valosin-containing protein (VCP) inhibitors. While all these studies reported target alterations via mutations in VCP as the primary mechanism of resistance, discrepancies persist to date regarding the zygosity of these mutations responsible for the resistance. In addition, the extent to which resistant cells harbor additional mutations in other genes is not well described. In this study, we performed global transcript analysis of the parental and previously reported VCP inhibitor (CB-5083) resistant cells and found additional mutations in the resistant cells. However, our CRISPR-Cas9 gene editing studies indicate that specific mutations in VCP are sufficient to produce resistance to CB-5083 suggesting the importance of on-target mutations in VCP for resistance. Strikingly, our analysis indicates a preexisting heterozygous frameshift mutation at codon 616 (N616fs*) in one of the VCP alleles in HCT116 cells, and we showed that this mutant allele is subjected to the nonsense-mediated decay (NMD). Accordingly, we identified a heterozygous mutation at codon 526 (L526S) in genomic DNA sequencing but a homozygous L526S mutation in complementary DNA sequencing in our independently generated CB-5083 resistant HCT116 cells, implying that the L526S mutation occurs in the allele that does not harbor the frameshift N616fs* mutation. Our results suggest the NMD as a possible mechanism for achieving the homozygosity of VCP mutant responsible for the resistance to VCP inhibitors while resolving the discrepancies among previous studies. Our results also underscore the importance of performing simultaneous genomic and complementary DNA sequencing when attributing mutational effects on the functionality particularly for an oligomer protein like VCP. Nature Publishing Group UK 2019-07-29 /pmc/articles/PMC6662852/ /pubmed/31358864 http://dx.doi.org/10.1038/s41598-019-47085-9 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Bastola, Prabhakar
Bilkis, Rabeya
De Souza, Cristabelle
Minn, Kay
Chien, Jeremy
Heterozygous mutations in valosin-containing protein (VCP) and resistance to VCP inhibitors
title Heterozygous mutations in valosin-containing protein (VCP) and resistance to VCP inhibitors
title_full Heterozygous mutations in valosin-containing protein (VCP) and resistance to VCP inhibitors
title_fullStr Heterozygous mutations in valosin-containing protein (VCP) and resistance to VCP inhibitors
title_full_unstemmed Heterozygous mutations in valosin-containing protein (VCP) and resistance to VCP inhibitors
title_short Heterozygous mutations in valosin-containing protein (VCP) and resistance to VCP inhibitors
title_sort heterozygous mutations in valosin-containing protein (vcp) and resistance to vcp inhibitors
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6662852/
https://www.ncbi.nlm.nih.gov/pubmed/31358864
http://dx.doi.org/10.1038/s41598-019-47085-9
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