Downregulation of miR‑98 contributes to hypoxic pulmonary hypertension by targeting ALK1

Li,Qingling; Zhou,Xincan; Zhou,Xianghui

doi:10.3892/mmr.2019.10482

Downregulation of miR‑98 contributes to hypoxic pulmonary hypertension by targeting ALK1

Authors:
- Qingling Li
- Xincan Zhou
- Xianghui Zhou
View Affiliations

Affiliations: Department of Respiratory Medicine, Xuzhou Municipal Hospital Affiliated to Xuzhou Medical University, Xuzhou, Jiangsu 221002, P.R. China
Published online on: July 9, 2019 https://doi.org/10.3892/mmr.2019.10482
Pages: 2167-2176
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Chronic hypoxia is one of the most common causes of secondary pulmonary hypertension, the mechanisms of which remain unclear. MicroRNAs (miRNAs) are small, noncoding RNAs that inhibit the translation or accelerate the degradation of mRNA. Previous studies have demonstrated that deregulated miRNA expression contributes to various cellular processes including cell apoptosis and proliferation, which are mediated by hypoxia. In the present study, the expression of miR‑98 was identified to be decreased in the lung tissue of a hypoxic pulmonary hypertension (HPH) rat model and pulmonary artery (PA) smooth muscle cells (PASMCs), which was induced by hypoxia. By transfecting miR‑98 mimics into PASMCs, the high expression of miR‑98 inhibited cell proliferation, but upregulated hypoxia‑induced PASMCs apoptosis. However, these effects of miR‑98 mimics on PASMCs were reversed by ALK1 (activin receptor‑like kinase‑1) overexpression. ALK1 was identified as a candidate target of miR‑98. In addition, overexpressing miR‑98 markedly decreased the pulmonary artery wall thickness and the right ventricular systolic pressure in rats induced by hypoxia. These results provided clear evidence that miR‑98 was a direct regulator of ALK1, and that the downregulation of miR‑98 contributed to the pathogenesis of HPH. These results provide a novel potential therapeutic strategy for the treatment of HPH.

View Figures

View References

1	Simonneau G, Gatzoulis MA, Adatia I, Celermajer D, Denton C, Ghofrani A, Gomez Sanchez MA, Kumar RK, Landzberg M, Machado RF, et al: Updated clinical classification of pulmonary hypertension. Turk Kardiyol Dern Arsivi (Turkish). 42 (Suppl):S45–S54. 2014.
2	D'Alonzo GE, Barst RJ, Ayres SM, Bergofsky EH, Brundage BH, Detre KM, Fishman AP, Goldring RM, Groves BM, Kernis JT, et al: Survival in patients with primary pulmonary hypertension. Results from a national prospective registry. Ann Intern Med. 115:343–349. 1991. View Article : Google Scholar : PubMed/NCBI
3	McLaughlin VV, Sitbon O, Badesch DB, Barst RJ, Black C, Galiè N, Rainisio M, Simonneau G and Rubin LJ: Survival with first-line bosentan in patients with primary pulmonary hypertension. Eur Respir J. 25:244–249. 2005. View Article : Google Scholar : PubMed/NCBI
4	Chan SY and Loscalzo J: Pathogenic mechanisms of pulmonary arterial hypertension. J Mol Cell Cardiol. 44:14–30. 2008. View Article : Google Scholar : PubMed/NCBI
5	Tang H, Desai AA and Yuan JX: Genetic insights into pulmonary arterial hypertension. Application of whole-exome sequencing to the study of pathogenic mechanisms. Am J Respir Crit Care Med. 194:393–397. 2016. View Article : Google Scholar : PubMed/NCBI
6	Zhou W, Negash S, Liu J and Raj JU: Modulation of pulmonary vascular smooth muscle cell phenotype in hypoxia: Role of cGMP-dependent protein kinase and myocardin. Am J Physiol Lung Cell Mol Physiol. 296:L780–L789. 2009. View Article : Google Scholar : PubMed/NCBI
7	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
8	Bushati N and Cohen SM: microRNA functions. Annu Rev Cell Dev Biol. 23:175–205. 2007. View Article : Google Scholar : PubMed/NCBI
9	Krol J, Loedige I and Filipowicz W: The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet. 11:597–610. 2010. View Article : Google Scholar : PubMed/NCBI
10	Bertero T, Lu Y, Annis S, Hale A, Bhat B, Saggar R, Saggar R, Wallace WD, Ross DJ, Vargas SO, et al: Systems-level regulation of microRNA networks by miR-130/301 promotes pulmonary hypertension. J Clin Invest. 124:3514–3528. 2014. View Article : Google Scholar : PubMed/NCBI
11	Caruso P, Dempsie Y, Stevens HC, McDonald RA, Long L, Lu R, White K, Mair KM, McClure JD, Southwood M, et al: A role for miR-145 in pulmonary arterial hypertension: Evidence from mouse models and patient samples. Circ Res. 111:290–300. 2012. View Article : Google Scholar : PubMed/NCBI
12	Potus F, Graydon C, Provencher S and Bonnet S: Vascular remodeling process in pulmonary arterial hypertension, with focus on miR-204 and miR-126 (2013 Grover Conference series). Pulm Circ. 4:175–184. 2014. View Article : Google Scholar : PubMed/NCBI
13	Gore B, Izikki M, Mercier O, Dewachter L, Fadel E, Humbert M, Dartevelle P, Simonneau G, Naeije R, Lebrin F and Eddahibi S: Key role of the endothelial TGF-beta/ALK1/endoglin signaling pathway in humans and rodents pulmonary hypertension. PLoS One. 9:e1003102014. View Article : Google Scholar : PubMed/NCBI
14	Savage-Dunn C: TGF-beta signaling. WormBook. 9:1–12. 2005.
15	Yan Y, Wang XJ, Li SQ, Yang SH, Lv ZC, Wang LT, He YY, Jiang X, Wang Y and Jing ZC: Elevated levels of plasma transforming growth factor-β1 in idiopathic and heritable pulmonary arterial hypertension. Int J Cardiol. 222:368–374. 2016. View Article : Google Scholar : PubMed/NCBI
16	Massague J: TGF-beta signal transduction. Ann Rev Biochem. 67:753–791. 1998. View Article : Google Scholar : PubMed/NCBI
17	Oh SP, Seki T, Goss KA, Imamura T, Yi Y, Donahoe PK, Li L, Miyazono K, ten Dijke P, Kim S and Li E: Activin receptor-like kinase 1 modulates transforming growth factor-beta 1 signaling in the regulation of angiogenesis. Proc Natl Acad Sci U S A. 97:2626–2631. 2000. View Article : Google Scholar : PubMed/NCBI
18	Uznanska-Loch B, Wiklo K, Kulczycka-Wojdala D, Szymańska B, Chrzanowski Ł, Wierzbowska-Drabik K, Trzos E, Kasprzak JD and Kurpesa M: Genetic variants in a Polish population of patients with pulmonary arterial hypertension: Sequencing of BMPR2, ALK1, and ENG genes. Kardiol Pol. 76:852–859. 2018. View Article : Google Scholar : PubMed/NCBI
19	Chida A, Shintani M, Yagi H, Fujiwara M, Kojima Y, Sato H, Imamura S, Yokozawa M, Onodera N, Horigome H, et al: Outcomes of childhood pulmonary arterial hypertension in BMPR2 and ALK1 mutation carriers. Am J Cardiol. 110:586–593. 2012. View Article : Google Scholar : PubMed/NCBI
20	Zhang H, Du L, Zhong Y, Flanders KC and Roberts JD Jr: Transforming growth factor-β stimulates Smad1/5 signaling in pulmonary artery smooth muscle cells and fibroblasts of the newborn mouse through ALK1. Am J Physiol Lung Cell Mol Physiol. 313:L615–L627. 2017. View Article : Google Scholar : PubMed/NCBI
21	Zhu D, Mackenzie NC, Shanahan CM, Shroff RC, Farquharson C and MacRae VE: BMP-9 regulates the osteoblastic differentiation and calcification of vascular smooth muscle cells through an ALK1 mediated pathway. J Cell Mol Med. 19:165–174. 2015. View Article : Google Scholar : PubMed/NCBI
22	Li L, Shi JY, Zhu GQ and Shi B: MiR-17-92 cluster regulates cell proliferation and collagen synthesis by targeting TGFB pathway in mouse palatal mesenchymal cells. J Cell Biochem. 113:1235–1244. 2012. View Article : Google Scholar : PubMed/NCBI
23	Wang X, Yan C, Xu X, Dong L, Su H, Hu Y, Zhang R and Ying K: Long noncoding RNA expression profiles of hypoxic pulmonary hypertension rat model. Gene. 579:23–28. 2016. View Article : Google Scholar : PubMed/NCBI
24	Krek A, Grun D, Poy MN, Wolf R, Rosenberg L, Epstein EJ, MacMenamin P, da Piedade I, Gunsalus KC, Stoffel M and Rajewsky N: Combinatorial microRNA target predictions. Nat Genet. 37:495–500. 2005. View Article : Google Scholar : PubMed/NCBI
25	Lewis BP, Shih IH, Jones-Rhoades MW, Bartel DP and Burge CB: Prediction of mammalian microRNA targets. Cell. 115:787–798. 2003. View Article : Google Scholar : PubMed/NCBI
26	Zhou S, Sun L, Cao C, Wu P, Li M, Sun G, Fei G, Ding X and Wang R: Hypoxia-induced microRNA-26b inhibition contributes to hypoxic pulmonary hypertension via CTGF. J Cell Biochem. 119:1942–1952. 2018. View Article : Google Scholar : PubMed/NCBI
27	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
28	Chen C, Ridzon DA, Broomer AJ, Zhou Z, Lee DH, Nguyen JT, Barbisin M, Xu NL, Mahuvakar VR, Andersen MR, et al: Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res. 33:e1792005. View Article : Google Scholar : PubMed/NCBI
29	Wang X: A PCR-based platform for microRNA expression profiling studies. RNA. 15:716–723. 2009. View Article : Google Scholar : PubMed/NCBI
30	Song Y, Jones JE, Beppu H, Keaney JF Jr, Loscalzo J and Zhang YY: Increased susceptibility to pulmonary hypertension in heterozygous BMPR2-mutant mice. Circulation. 112:553–562. 2005. View Article : Google Scholar : PubMed/NCBI
31	Fischer AH, Jacobson KA, Rose J and Zeller R: Hematoxylin and eosin staining of tissue and cell sections. CSH Protoc 2008: pdb prot4986. 2008.
32	Xu X, Hu H, Wang X, Ye W, Su H, Hu Y, Dong L, Zhang R and Ying K: Involvement of CapG in proliferation and apoptosis of pulmonary arterial smooth muscle cells and in hypoxia-induced pulmonary hypertension rat model. Exp Lung Res. 42:142–153. 2016. View Article : Google Scholar : PubMed/NCBI
33	Thum T, Galuppo P, Wolf C, Fiedler J, Kneitz S, van Laake LW, Doevendans PA, Mummery CL, Borlak J, Haverich A, et al: MicroRNAs in the human heart: A clue to fetal gene reprogramming in heart failure. Circulation. 116:258–267. 2007. View Article : Google Scholar : PubMed/NCBI
34	Mann DL: MicroRNAs and the failing heart. N Eng J Med. 356:2644–2645. 2007. View Article : Google Scholar
35	Sarkar J, Gou D, Turaka P, Viktorova E, Ramchandran R and Raj JU: MicroRNA-21 plays a role in hypoxia-mediated pulmonary artery smooth muscle cell proliferation and migration. Am J Physiol Lung Cell Mol Physiol. 299:L861–L871. 2010. View Article : Google Scholar : PubMed/NCBI
36	Courboulin A, Paulin R, Giguère NJ, Saksouk N, Perreault T, Meloche J, Paquet ER, Biardel S, Provencher S, Côté J, et al: Role for miR-204 in human pulmonary arterial hypertension. J Exp Med. 208:535–548. 2011. View Article : Google Scholar : PubMed/NCBI
37	Guo L, Qiu Z, Wei L, Yu X, Gao X, Jiang S, Tian H, Jiang C and Zhu D: The microRNA-328 regulates hypoxic pulmonary hypertension by targeting at insulin growth factor 1 receptor and L-type calcium channel-α1C. Hypertension. 59:1006–1013. 2012. View Article : Google Scholar : PubMed/NCBI
38	Siragam V, Rutnam ZJ, Yang W, Fang L, Luo L, Yang X, Li M, Deng Z, Qian J, Peng C and Yang BB: MicroRNA miR-98 inhibits tumor angiogenesis and invasion by targeting activin receptor-like kinase-4 and matrix metalloproteinase-11. Oncotarget. 3:1370–1385. 2012. View Article : Google Scholar : PubMed/NCBI
39	Trembath RC: Mutations in the TGF-beta type 1 receptor, ALK1, in combined primary pulmonary hypertension and hereditary haemorrhagic telangiectasia, implies pathway specificity. J Heart Lung Transplant. 20:1752001. View Article : Google Scholar : PubMed/NCBI
40	Fujiwara M, Yagi H, Matsuoka R, Akimoto K, Furutani M, Imamura S, Uehara R, Nakayama T, Takao A, Nakazawa M and Saji T: Implications of mutations of activin receptor-like kinase 1 gene (ALK1) in addition to bone morphogenetic protein receptor II gene (BMPR2) in children with pulmonary arterial hypertension. Circ J. 72:127–133. 2008. View Article : Google Scholar : PubMed/NCBI
41	Jerkic M, Kabir MG, Davies A, Yu LX, McIntyre BA, Husain NW, Enomoto M, Sotov V, Husain M, Henkelman M, et al: Pulmonary hypertension in adult Alk1 heterozygous mice due to oxidative stress. Cardiovasc Res. 92:375–384. 2011. View Article : Google Scholar : PubMed/NCBI
42	Soubrier F, Chung WK, Machado R, Grünig E, Aldred M, Geraci M, Loyd JE, Elliott CG, Trembath RC, Newman JH and Humbert M: Genetics and genomics of pulmonary arterial hypertension. J Am Coll Cardiol. 62:D13–D21. 2013. View Article : Google Scholar : PubMed/NCBI