Pri-microRNA-124 rs531564 polymorphism minor allele increases the risk of pulmonary artery hypertension by abnormally enhancing proliferation of pulmonary artery smooth muscle cells

Abstract

MicroRNA-124 (miR-124) has been reported to be downregulated in the cells exposed to hypoxia, which was confirmed in our study. We then used online microRNA target prediction tools to identify GRB2, SMAD5, and JAG1 as the candidate target genes of miR-124, and we next validated GRB2 as a direct gene by using luciferase reporter system. We also established the regulatory relationship between miR-124 and GRB2 by showing the negative linear relationship between GRB2 and miR-124 expression. Furthermore, we investigated the miR-124 and GRB2 expression levels of different genotypes including CC (n=30), GC (n=18), and GG (n=4), which supported the hypothesis that the presence of minor allele (C) of rs531564 polymorphism compromised the expression of miR-124. Meanwhile, we also conducted real-time polymerase chain reaction and Western blot analysis to study the expression of GRB2 among different genotypes or pulmonary artery smooth muscle cells (PASMCs) treated with miR-124 mimics, GRB2 small interfering RNA, and miR-124 inhibitors, respectively, and found that introduction of miR-124 or GRB2 small interfering RNA could reduce the expression of GRB2 and inhibit the proliferation of PASMCs, while miR-124 upregulated the expression of GRB2 and promoted the proliferation of PASMCs. A total of 412 COPD patients with PAH (n=182) or without PAH (n=230) were recruited in this study, and more individuals carrying at least one minor allele of rs531564 were found in the COPD patients with PAH than in those without PAH (odds ratio: 0.61, 95% confidence interval: 0.41–0.91; P=0.166). In conclusion, the presence of rs531564 minor allele may increase the risk of PAH in COPD by reducing miR-124 expression, increasing GRB2 expression, and promoting the proliferation of PASMCs.

Circulating miRNAs as molecular markers of occupational grain dust exposure

“…In our miRNA-disease association analysis, coronary artery disease was one of the disease terms associated with has-miR-124-3p. In human pulmonary artery smooth muscle cells www.nature.com/scientificreports/ in vitro, hsa-miR-124-3p was shown to target GRB2 34 . The ERK2/GRB2/Shc pathway is critical in vascular smooth muscle cell proliferation, and this indicates that differential expression of hsa-miR-124-3p might be involved in the pathogenesis of pulmonary artery hypertension in patients with COPD.…”

“…Age (year) a 45 49 Height (cm) a 180 178 Weight (kg) a 89 (63-121) 90 Body mass index (kg/m 2 ) a 27 (22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36) 27 (23)(24)(25)(26)(27)(28)(29)(30)(31)(32) Cold (in %) 19 27 Farm childhood (in %) 36 46…”

Correlation between pri-miR-124 (rs531564) polymorphism and congenital heart disease susceptibility in Chinese population at two different altitudes: a case-control and in silico study

“…In addition, previous studies have suggested that hypoxia induces miR-124 downregulation which promotes prostate cancer cell survival, leads to chondrogenesis, and promotes proliferation of pulmonary artery smooth muscle cells. These functions are mediated by the regulation of PIM1 , NFATc1 , and GRB2 , respectively (Gong et al 2017; Gu et al 2016; Li et al 2017). The downregulated miR-124 also exhibits a cardio-protective effect by attenuating endoplasmic reticulum stress (Bao et al 2017).…”

“…Hu et al 2010 reported that upregulated hypoxia-responsive miR-210 exhibits a cardio-protective effect by inhibiting Efna3 and Ptp1b , in a mouse model of myocardial infarction. Besides, it is widely reported that miR-124 is a downregulated hypoxia-responsive miRNA involved in the regulation of biological processes in a variety of cells (Gong et al 2017; Gu et al 2016; Li et al 2017). Moreover, a line of evidence has demonstrated that rs531564 (G>C), a functional SNP of the primary miR-124 ( pri-miR-124 ) sequence, is associated with the expression of mature miR-124.…”

“…Moreover, a line of evidence has demonstrated that rs531564 (G>C), a functional SNP of the primary miR-124 ( pri-miR-124 ) sequence, is associated with the expression of mature miR-124. The G allele carriers, which represent the major population, have a higher expression level of miR-124 transcript than the C allele carriers, which represent a minor population (Chen et al 2018; Li et al 2017; Zou et al 2017). The rs531564 polymorphism of pri-miR-124 is also one of the risk factors for human cervical cancer, esophageal squamous cell carcinoma, colorectal cancer, and type 2 diabetes (Gao et al 2015; Li et al 2015; Wu and Zhang 2014; Ye et al 2008).…”