Assessment of microRNA and gene dysregulation in pulmonary hypertension by endoarterial biopsy

Abstract

MicroRNAs (miRNAs) may regulate a number of genes, each of which may have a variety of functions. We utilized an endoarterial biopsy catheter to assess the dysregulation of miRNAs in a porcine shunt model of pulmonary hypertension (PH). Two Yucatan micropigs underwent surgical anastomosis of the left pulmonary artery to the descending aorta. Endoarterial biopsy samples were obtained at baseline, and at regular intervals during the progression of PH. RNA, isolated from biopsy samples, was analyzed by Illumina miRNA expression microarrays (containing ∼1200 human miRNAs), Affymetrix Porcine GeneChips, Bioconductor, and GeneSpring. We examined a total of 925 genes in a PH whole genome microarray. Biopsy samples showed that 39 miRNAs were downregulated and 34 miRNAs were upregulated compared to baseline. The number of PH-associated genes reported to be controlled by each of the dysregulated miRNAs was in the range of 1–113. The five miRNAs that had the largest number of PH-associated genes were: miR-548c-3p, miR-520d-3p, miR-130a-5p, miR-30a-3p, and miR-let-7g-3p. Several of the dysregulated miRNAs have been associated with molecular pathways and biologic processes involved in PH. Among 29 miRNAs, which were predicted to be dysregulated by a systems biology approach, we found four that were dysregulated in our porcine shunt model. An endoarterial biopsy technique was successful in showing that a large number of miRNAs are dysregulated in a porcine shunt model of PH. Many of these miRNAs control multiple PH-associated genes, molecular pathways, and biologic processes. Endoarterial biopsy offers potential experimental and clinical diagnostic value.