Scientific Reports (May 2017)

Molecular and functional characterization of the BMPR2 gene in Pulmonary Arterial Hypertension

  • Guillermo Pousada,
  • Vincenzo Lupo,
  • Sheila Cástro-Sánchez,
  • María Álvarez-Satta,
  • Ana Sánchez-Monteagudo,
  • Adolfo Baloira,
  • Carmen Espinós,
  • Diana Valverde

DOI
https://doi.org/10.1038/s41598-017-02074-8
Journal volume & issue
Vol. 7, no. 1
pp. 1 – 19

Abstract

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Abstract Pulmonary arterial hypertension is a progressive disease that causes the obstruction of precapillary pulmonary arteries and a sustained increase in pulmonary vascular resistance. The aim was to analyze functionally the variants found in the BMPR2 gene and to establish a genotype-phenotype correlation. mRNA expression studies were performed using pSPL3 vector, studies of subcellular localization were performed using pEGFP-N1 vector and luciferase assays were performed using pGL3-Basic vector. We have identified 30 variants in the BMPR2 gene in 27 of 55 patients. In 16 patients we detected pathogenic mutations. Minigene assays revealed that 6 variants (synonymous, missense) result in splicing defect. By immunofluorescence assay, we observed that 4 mutations affect the protein localization. Finally, 4 mutations located in the 5′UTR region showed a decreased transcriptional activity in luciferase assays. Genotype-phenotype correlation, revealed that patients with pathogenic mutations have a more severe phenotype (sPaP p = 0.042, 6MWT p = 0.041), a lower age at diagnosis (p = 0.040) and seemed to have worse response to phosphodiesterase-5-inhibitors (p = 0.010). Our study confirms that in vitro expression analysis is a suitable approach in order to investigate the phenotypic consequences of the nucleotide variants, especially in cases where the involved genes have a pattern of expression in tissues of difficult access.