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Identification of Novel Mutations and Expressions of EPAS1 in Phaeochromocytomas and Paragangliomas

Endothelial PAS domain-containing protein 1 (EPAS1) is an oxygen-sensitive component of the hypoxia-inducible factors (HIFs) having reported implications in many cancers by inducing a pseudo-hypoxic microenvironment. However, the molecular dysregulation and clinical significance of EPAS1 has never b...

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Autores principales: Islam, Farhadul, Pillai, Suja, Gopalan, Vinod, Lam, Alfred King-Yin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7693385/
https://www.ncbi.nlm.nih.gov/pubmed/33114456
http://dx.doi.org/10.3390/genes11111254
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author Islam, Farhadul
Pillai, Suja
Gopalan, Vinod
Lam, Alfred King-Yin
author_facet Islam, Farhadul
Pillai, Suja
Gopalan, Vinod
Lam, Alfred King-Yin
author_sort Islam, Farhadul
collection PubMed
description Endothelial PAS domain-containing protein 1 (EPAS1) is an oxygen-sensitive component of the hypoxia-inducible factors (HIFs) having reported implications in many cancers by inducing a pseudo-hypoxic microenvironment. However, the molecular dysregulation and clinical significance of EPAS1 has never been investigated in depth in phaeochromocytomas/paragangliomas. This study aims to identify EPAS1 mutations and alterations in DNA copy number, mRNA and protein expression in patients with phaeochromocytomas/paragangliomas. The association of molecular dysregulations of EPAS1 with clinicopathological factors in phaeochromocytomas and paragangliomas were also analysed. High-resolution melt-curve analysis followed by Sanger sequencing was used to detect mutations in EPAS1. EPAS1 DNA number changes and mRNA expressions were examined by polymerase chain reaction (PCR). Immunofluorescence assay was used to study EPAS1 protein expression. In phaeochromocytomas, 12% (n = 7/57) of patients had mutations in the EPAS1 sequence, which includes two novel mutations (c.1091A>T; p.Lys364Met and c.1129A>T; p.Ser377Cys). Contrastingly, in paragangliomas, 7% (n = 1/14) of patients had EPAS1 mutations and only the c.1091A>T; p.Lys364Met mutation was detected. In silico analysis revealed that the p.Lys364Met mutation has pathological potential based on the functionality of the protein, whereas the p.Ser377Cys mutation was predicted to be neutral or tolerated. The majority of the patients had EPAS1 DNA amplification (79%; n = 56/71) and 53% (n = 24/45) patients shown mRNA overexpression. Most of the patients with EPAS1 mutations exhibited aberrant DNA changes, mRNA and protein overexpression. In addition, these alterations of EPAS1 were associated with tumour weight and location. Thus, the molecular dysregulation of EPAS1 could play crucial roles in the pathogenesis of phaeochromocytomas and paragangliomas.
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spelling pubmed-76933852020-11-28 Identification of Novel Mutations and Expressions of EPAS1 in Phaeochromocytomas and Paragangliomas Islam, Farhadul Pillai, Suja Gopalan, Vinod Lam, Alfred King-Yin Genes (Basel) Article Endothelial PAS domain-containing protein 1 (EPAS1) is an oxygen-sensitive component of the hypoxia-inducible factors (HIFs) having reported implications in many cancers by inducing a pseudo-hypoxic microenvironment. However, the molecular dysregulation and clinical significance of EPAS1 has never been investigated in depth in phaeochromocytomas/paragangliomas. This study aims to identify EPAS1 mutations and alterations in DNA copy number, mRNA and protein expression in patients with phaeochromocytomas/paragangliomas. The association of molecular dysregulations of EPAS1 with clinicopathological factors in phaeochromocytomas and paragangliomas were also analysed. High-resolution melt-curve analysis followed by Sanger sequencing was used to detect mutations in EPAS1. EPAS1 DNA number changes and mRNA expressions were examined by polymerase chain reaction (PCR). Immunofluorescence assay was used to study EPAS1 protein expression. In phaeochromocytomas, 12% (n = 7/57) of patients had mutations in the EPAS1 sequence, which includes two novel mutations (c.1091A>T; p.Lys364Met and c.1129A>T; p.Ser377Cys). Contrastingly, in paragangliomas, 7% (n = 1/14) of patients had EPAS1 mutations and only the c.1091A>T; p.Lys364Met mutation was detected. In silico analysis revealed that the p.Lys364Met mutation has pathological potential based on the functionality of the protein, whereas the p.Ser377Cys mutation was predicted to be neutral or tolerated. The majority of the patients had EPAS1 DNA amplification (79%; n = 56/71) and 53% (n = 24/45) patients shown mRNA overexpression. Most of the patients with EPAS1 mutations exhibited aberrant DNA changes, mRNA and protein overexpression. In addition, these alterations of EPAS1 were associated with tumour weight and location. Thus, the molecular dysregulation of EPAS1 could play crucial roles in the pathogenesis of phaeochromocytomas and paragangliomas. MDPI 2020-10-24 /pmc/articles/PMC7693385/ /pubmed/33114456 http://dx.doi.org/10.3390/genes11111254 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
Islam, Farhadul
Pillai, Suja
Gopalan, Vinod
Lam, Alfred King-Yin
Identification of Novel Mutations and Expressions of EPAS1 in Phaeochromocytomas and Paragangliomas
title Identification of Novel Mutations and Expressions of EPAS1 in Phaeochromocytomas and Paragangliomas
title_full Identification of Novel Mutations and Expressions of EPAS1 in Phaeochromocytomas and Paragangliomas
title_fullStr Identification of Novel Mutations and Expressions of EPAS1 in Phaeochromocytomas and Paragangliomas
title_full_unstemmed Identification of Novel Mutations and Expressions of EPAS1 in Phaeochromocytomas and Paragangliomas
title_short Identification of Novel Mutations and Expressions of EPAS1 in Phaeochromocytomas and Paragangliomas
title_sort identification of novel mutations and expressions of epas1 in phaeochromocytomas and paragangliomas
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7693385/
https://www.ncbi.nlm.nih.gov/pubmed/33114456
http://dx.doi.org/10.3390/genes11111254
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