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Circulation. 2018 Jul 17;138(3):287-304. doi: 10.1161/CIRCULATIONAHA.117.031258. Epub 2018 Feb 5.

Epigenetic Dysregulation of the Dynamin-Related Protein 1 Binding Partners MiD49 and MiD51 Increases Mitotic Mitochondrial Fission and Promotes Pulmonary Arterial Hypertension: Mechanistic and Therapeutic Implications.

Author information

1
Department of Medicine (K.-H.C., A.D., F.P., J.F., J.M., D.W., K.D.-S., A.L.T., M.L.O., S.L.A.).
2
Queen's University, Kingston, Canada. Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, UK (J.L., J.I, A.L.).
3
Pulmonary Hypertension Research Group of the University Cardiology and Pulmonary Institute of the Quebec Research Centre, Laval University, Quebec City, Canada (S.B.).
4
Queen's Cardiopulmonary Unit, Translational Institute of Medicine, Department of Medicine (J.M., C.H., M.L.O., S.L.A.).
5
State Key Laboratory of Cardiovascular Disease, FuWai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Z.-C.J.).
6
Department of Medicine (K.-H.C., A.D., F.P., J.F., J.M., D.W., K.D.-S., A.L.T., M.L.O., S.L.A.) stephen.archer@queensu.ca.

Abstract

BACKGROUND:

Mitotic fission is increased in pulmonary arterial hypertension (PAH), a hyperproliferative, apoptosis-resistant disease. The fission mediator dynamin-related protein 1 (Drp1) must complex with adaptor proteins to cause fission. Drp1-induced fission has been therapeutically targeted in experimental PAH. Here, we examine the role of 2 recently discovered, poorly understood Drp1 adapter proteins, mitochondrial dynamics protein of 49 and 51 kDa (MiD49 and MiD51), in normal vascular cells and explore their dysregulation in PAH.

METHODS:

Immunoblots of pulmonary artery smooth muscle cells (control, n=6; PAH, n=8) and immunohistochemistry of lung sections (control, n=6; PAH, n=6) were used to assess the expression of MiD49 and MiD51. The effects of manipulating MiDs on cell proliferation, cell cycle, and apoptosis were assessed in human and rodent PAH pulmonary artery smooth muscle cells with flow cytometry. Mitochondrial fission was studied by confocal imaging. A microRNA (miR) involved in the regulation of MiD expression was identified using microarray techniques and in silico analyses. The expression of circulatory miR was assessed with quantitative reverse transcription-polymerase chain reaction in healthy volunteers (HVs) versus patients with PAH from Sheffield, UK (plasma: HV, n=29, PAH, n=27; whole blood: HV, n=11, PAH, n=14) and then confirmed in a cohort from Beijing, China (plasma: HV, n=19, PAH, n=36; whole blood: HV, n=20, PAH, n=39). This work was replicated in monocrotaline and Sugen 5416-hypoxia, preclinical PAH models. Small interfering RNAs targeting MiDs or an miR mimic were nebulized to rats with monocrotaline-induced PAH (n=4-10).

RESULTS:

MiD expression is increased in PAH pulmonary artery smooth muscle cells, which accelerates Drp1-mediated mitotic fission, increases cell proliferation, and decreases apoptosis. Silencing MiDs (but not other Drp1 binding partners, fission 1 or mitochondrial fission factor) promotes mitochondrial fusion and causes G1-phase cell cycle arrest through extracellular signal-regulated kinases 1/2- and cyclin-dependent kinase 4-dependent mechanisms. Augmenting MiDs in normal cells causes fission and recapitulates the PAH phenotype. MiD upregulation results from decreased miR-34a-3p expression. Circulatory miR-34a-3p expression is decreased in both patients with PAH and preclinical models of PAH. Silencing MiDs or augmenting miR-34a-3p regresses experimental PAH.

CONCLUSIONS:

In health, MiDs regulate Drp1-mediated fission, whereas in disease, epigenetic upregulation of MiDs increases mitotic fission, which drives pathological proliferation and apoptosis resistance. The miR-34a-3p-MiD pathway offers new therapeutic targets for PAH.

KEYWORDS:

cell cycle; cyclin-dependent kinases; dynamin-related protein 1; microRNAs; mitochondrial dynamics; mitochondrial fission; pulmonary arterial hypertension

PMID:
29431643
PMCID:
PMC6050130
[Available on 2019-07-17]
DOI:
10.1161/CIRCULATIONAHA.117.031258
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