Characterization of rare ABCC8 variants identified in Spanish pulmonary arterial hypertension patients.
Sci Rep
; 10(1): 15135, 2020 09 15.
Article
em En
| MEDLINE
| ID: mdl-32934261
ABSTRACT
Pulmonary Arterial Hypertension (PAH) is a rare and fatal disease where knowledge about its genetic basis continues to increase. In this study, we used targeted panel sequencing in a cohort of 624 adult and pediatric patients from the Spanish PAH registry. We identified 11 rare variants in the ATP-binding Cassette subfamily C member 8 (ABCC8) gene, most of them with splicing alteration predictions. One patient also carried another variant in SMAD1 gene (c.27delinsGTAAAG). We performed an ABCC8 in vitro biochemical analyses using hybrid minigenes to confirm the correct mRNA processing of 3 missense variants (c.211C > T p.His71Tyr, c.298G > A p.Glu100Lys and c.1429G > A p.Val477Met) and the skipping of exon 27 in the novel splicing variant c.3394G > A. Finally, we used structural protein information to further assess the pathogenicity of the variants. The results showed 11 novel changes in ABCC8 and 1 in SMAD1 present in PAH patients. After in silico and in vitro biochemical analyses, we classified 2 as pathogenic (c.3288_3289del and c.3394G > A), 6 as likely pathogenic (c.211C > T, c.1429G > A, c.1643C > T, c.2422C > A, c.2694 + 1G > A, c.3976G > A and SMAD1 c.27delinsGTAAAG) and 3 as Variants of Uncertain Significance (c.298G > A, c.2176G > A and c.3238G > A). In all, we show that coupling in silico tools with in vitro biochemical studies can improve the classification of genetic variants.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Marcadores Genéticos
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Splicing de RNA
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Éxons
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Mutação de Sentido Incorreto
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Receptores de Sulfonilureias
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Hipertensão Arterial Pulmonar
Tipo de estudo:
Incidence_studies
/
Prognostic_studies
Limite:
Adult
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Female
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Humans
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Male
País como assunto:
Europa
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article