Novel Loss-of-Function Variants in Pulmonary Arterial Hypertension.

Alba Vera-Zambrano, Mauro Lago-Docampo, Natalia Gallego, Juan Felipe Franco-Gonzalez, Daniel Morales-Cano, Alejandro Cruz-Utrilla, Marta Villegas-Esguevillas, Edgar Fernández-Malavé, Pilar Escribano-Subías, Jair Antonio Tenorio-Castaño, Francisco Perez-Vizcaino, Diana Valverde, Teresa González, Angel Cogolludo

Journal: American journal of respiratory cell and molecular biology 2023;69(2):147-158

PMID: 36917789

Full Text on Atypon View on PubMed

Abstract

Reduced expression and/or activity of Kv1.5 channels (encoded by ) is a common hallmark in human or experimental pulmonary arterial hypertension (PAH). Likewise, genetic variants in have been found in patients with PAH, but their functional consequences and potential impact on the disease are largely unknown. Herein, this study aimed to characterize the functional consequences of seven variants found in a cohort of patients with PAH. Potassium currents were recorded by patch-clamp technique in HEK293 cells transfected with wild-type or mutant Kv1.5 cDNA. Flow cytometry, Western blot, and confocal microscopy techniques were used for measuring protein expression and cell apoptosis in HEK293 and human pulmonary artery smooth muscle cells. variants (namely, Arg184Pro and Gly384Arg) found in patients with PAH resulted in a clear loss of potassium channel function as assessed by electrophysiological and molecular modeling analyses. The Arg184Pro variant also resulted in a pronounced reduction of Kv1.5 expression. Transfection with Arg184Pro or Gly384Arg variants decreased apoptosis of human pulmonary artery smooth muscle cells compared with the wild-type cells, demonstrating that dysfunction in both variants affects cell viability. Thus, in addition to affecting channel activity, both variants were associated with impaired apoptosis, a crucial process linked to the disease. The estimated prevalence of dysfunctional variants in the PAH population analyzed was around 1%. The data indicate that some variants found in patients with PAH have critical consequences for channel function, supporting the idea that pathogenic variants may be a causative or contributing factor for PAH.

Address: Department of Pharmacology and Toxicology and.; Department of Biochemistry, School of Medicine, University Autonoma of Madrid, Madrid, Spain.; Instituto de Investigaciones Biomédicas "Alberto Sols" Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, Madrid, Spain.; Centro de Investigación en Nanomateriais e Biomedicina (CINBIO), Universidade de Vigo, Vigo, Spain.; Rare Diseases and Pediatric Medicine, Galicia Sur Health Research Institute, SERGAS-UVIGO, Vigo, Spain.; Institute of Medical and Molecular Genetics-IdiPAZ, Universitary Hospital La Paz, University Autonoma of Madrid, Madrid, Spain.; Centro de Investigación Biomédica en Red de Enfermedades Raras, Instituto de Salud Carlos III, Madrid, Spain.; ITHACA, European Reference Network on Rare Congenital Malformations and Rare Intellectual Disability, University Hospital La Paz, Madrid, Spain.; Department of Structural and Chemical Biology, Margarita Salas Center for Biological Research, Scientific Research Council, Madrid, Spain.; Experimental Pathology of Atherosclerosis Laboratory, National Center for Cardiovascular Research Carlos III, Madrid, Spain.; Atherosclerosis Research Unit, Department of Clinical Medicine, Aarhus University, Aarhus, Denmark.; Centro de Investigación Biomédica en Red de Enfermedades Cardiovasculares, and.; Unidad Multidisciplinar de Hipertensión Pulmonar, Servicio de Cardiología, Hospital Universitario 12 de Octubre, Madrid, Spain.; Centro de Investigación Biomédica en Red de Enfermedades Respitatorias, Instituto de Salud Carlos III, Madrid, Spain.; Instituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain; and.; Department of Immunology, Ophthalmology and ENT, School of Medicine, University Complutense of Madrid, Madrid, Spain.; Instituto de Investigación Sanitaria Hospital 12 de Octubre, Madrid, Spain.

DOI: 10.1165/rcmb.2022-0245OC

Link outs

Free resources

Medical:

https://medlineplus.gov/pulmonaryhypertension.html

Molecular Biology Databases:

https://glygen.org/publication/PubMed/36917789/

Subscription / membership required

Full Text Sources:

MeSH Terms: Humans, Pulmonary Arterial Hypertension, HEK293 Cells, Hypertension, Pulmonary, Kv1.5 Potassium Channel, Familial Primary Pulmonary Hypertension, Pulmonary Artery