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Circulation. 2017 Dec 19;136(25):2451-2467. doi: 10.1161/CIRCULATIONAHA.117.028034. Epub 2017 Sep 26.

Identification of MicroRNA-124 as a Major Regulator of Enhanced Endothelial Cell Glycolysis in Pulmonary Arterial Hypertension via PTBP1 (Polypyrimidine Tract Binding Protein) and Pyruvate Kinase M2.

Author information

1
Division of Respiratory Medicine, Department of Medicine, University of Cambridge School of Clinical Medicine, Addenbrooke's Hospital, United Kingdom (P.C., B.J.D., L.L., P.D.U., C.H., F.N.K., L.A.H.., N.W.M.) pc468@medschl.cam.ac.uk nwm23@medschl.cam.ac.uk.
2
Division of Respiratory Medicine, Department of Medicine, University of Cambridge School of Clinical Medicine, Addenbrooke's Hospital, United Kingdom (P.C., B.J.D., L.L., P.D.U., C.H., F.N.K., L.A.H.., N.W.M.).
3
Ottawa Hospital Research Institute and University of Ottawa, Ontario, Canada (K.S., C.P.-I., J.R.L., M.T., D.J.S.).
4
Laboratory of Angiogenesis and Neurovascular Link, Vesalius Research Center, VIB, Leuven, Belgium (S.S., P.C.).
5
Laboratory of Angiogenesis and Neurovascular Link, Vesalius Research Center, Department of Oncology, University of Leuven, Belgium (S.S., P.C.).
6
Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Ontario, Canada (C.D.S., M.L.O.).
7
Cardiovascular Pulmonary Research Laboratories, Departments of Pediatrics and Medicine (H.Z., A.R.F., M.G.F., K.R.S.).
8
Department of Biochemistry and Molecular Genetics (A.D.).
9
University of Colorado, Anschutz Medical Campus, Aurora. Cardiovascular Research Institute, University of California, San Francisco (A.H.).

Abstract

BACKGROUND:

Pulmonary arterial hypertension (PAH) is characterized by abnormal growth and enhanced glycolysis of pulmonary artery endothelial cells. However, the mechanisms underlying alterations in energy production have not been identified.

METHODS:

Here, we examined the miRNA and proteomic profiles of blood outgrowth endothelial cells (BOECs) from patients with heritable PAH caused by mutations in the bone morphogenetic protein receptor type 2 (BMPR2) gene and patients with idiopathic PAH to determine mechanisms underlying abnormal endothelial glycolysis. We hypothesized that in BOECs from patients with PAH, the downregulation of microRNA-124 (miR-124), determined with a tiered systems biology approach, is responsible for increased expression of the splicing factor PTBP1 (polypyrimidine tract binding protein), resulting in alternative splicing of pyruvate kinase muscle isoforms 1 and 2 (PKM1 and 2) and consequently increased PKM2 expression. We questioned whether this alternative regulation plays a critical role in the hyperglycolytic phenotype of PAH endothelial cells.

RESULTS:

Heritable PAH and idiopathic PAH BOECs recapitulated the metabolic abnormalities observed in pulmonary artery endothelial cells from patients with idiopathic PAH, confirming a switch from oxidative phosphorylation to aerobic glycolysis. Overexpression of miR-124 or siRNA silencing of PTPB1 restored normal proliferation and glycolysis in heritable PAH BOECs, corrected the dysregulation of glycolytic genes and lactate production, and partially restored mitochondrial respiration. BMPR2 knockdown in control BOECs reduced the expression of miR-124, increased PTPB1, and enhanced glycolysis. Moreover, we observed reduced miR-124, increased PTPB1 and PKM2 expression, and significant dysregulation of glycolytic genes in the rat SUGEN-hypoxia model of severe PAH, characterized by reduced BMPR2 expression and endothelial hyperproliferation, supporting the relevance of this mechanism in vivo.

CONCLUSIONS:

Pulmonary vascular and circulating progenitor endothelial cells isolated from patients with PAH demonstrate downregulation of miR-124, leading to the metabolic and proliferative abnormalities in PAH ECs via PTPB1 and PKM1/PKM2. Therefore, the manipulation of this miRNA or its targets could represent a novel therapeutic approach for the treatment of PAH.

KEYWORDS:

endothelial cells; endothelial progenitor cells; glycolysis; hypertension, pulmonary; metabolism; microRNAs

PMID:
28971999
PMCID:
PMC5736425
[Available on 2018-12-19]
DOI:
10.1161/CIRCULATIONAHA.117.028034
[Indexed for MEDLINE]

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