Metabolic alterations in human pulmonary artery smooth muscle cells treated with PDGF-BB

Abstract

Background: Metabolic abnormalities are considered to play a key regulatory role in vascular remodeling of pulmonary arterial hypertension. However, to date, there is a paucity of research documenting the changes in metabolome profiles within the supernatants of pulmonary artery smooth muscle cells (PASMC) during their transition from a contractile to a synthetic phenotype.

Methods: CCK-8 and Edu staining assays were used to evaluate the cell viability and proliferation of human PASMCs. IncuCyte ZOOM imaging system was used to continuously and automatically detect the migration of the PASMCs. A targeted metabolomics profiling was performed to quantitatively analyze 121 metabolites in the supernatant. Orthogonal partial least squares discriminant analysis was used to discriminate between PDGF-BB-induced PASMCs and controls. Metabolite set enrichment analysis was adapted to exploit the most disturbed metabolic pathways.

Results: Human PASMCs exhibited a transformation from contractile phenotype to synthetic phenotype after PDGF-BB induction, along with a significant increase in cell viability, proliferation, and migration. Metabolites in the supernatants of PASMCs treated with or without PDGF-BB were well profiled. Eleven metabolites were found to be significantly upregulated, whereas seven metabolites were downregulated in the supernatants of PASMCs induced by PDGF-BB compared to the vehicle-treated cells. Fourteen pathways were involved, and pyruvate metabolism pathway was ranked first with the highest enrichment impact followed by glycolysis/gluconeogenesis and pyrimidine metabolism.

Conclusions: Significant and extensive metabolic abnormalities occurred during the phenotypic transformation of PASMCs. Disturbance of pyruvate metabolism pathway might contribute to pulmonary vascular remodeling.

Keywords

metabolomics / pulmonary artery hypertension / pyruvate metabolism / vascular remodeling

References

[1] Humbert M, Kovacs G, Hoeper MM. Et al; ESC/ERS scientific document group. 2022 ESC/ERS guidelines for the diagnosis and treatment of pulmonary hypertension. Eur Heart J. 2022; 43(38): 3618-3731.

[2] Hassoun PM. Pulmonary arterial Hypertension. N Engl J Med. 2021; 385(25): 2361-2376.

[3] Poch D, Mandel J. Pulmonary hypertension. Ann Intern Med. 2021; 174: ITC49-ITC64.

[4] Benza RL, Miller DP, Barst RJ, Badesch DB, Frost AE, McGoon MD. An evaluation of long-term survival from time of diagnosis in pulmonary arterial hypertension from the REVEAL registry. Chest. 2012; 142(2): 448-456.

[5] McLaughlin VV, Hoeper MM, Channick RN, et al. Pulmonary arterial Hypertension-related morbidity is prognostic for mortality. J Am Coll Cardiol. 2018; 71(7): 752-763.

[6] Wu XH, He YY, Chen ZR, et al. Single-cell analysis of peripheral blood from high-altitude pulmonary hypertension patients identifies a distinct monocyte phenotype. Nat Commun. 2023; 14(1): 1820.

[7] Yan Y, He YY, Jiang X, et al. DNA methyltransferase 3B deficiency unveils a new pathological mechanism of pulmonary hypertension. Sci Adv. 2020; 6(50): eaba2470.

[8] Liu SF, Yan Y. Animal models of pulmonary hypertension due to left heart disease. Animal Model Exp Med. 2022; 5: 197-206.

[9] Liu Y, Shi JZ, Jiang R, et al. Regulatory T cell-related gene indicators in pulmonary Hypertension. Front Pharmacol. 2022; 31(13): 908783.

[10] Lewis GD, Ngo D, Hemnes AR, et al. Metabolic profiling of right ventricular-pulmonary vascular function reveals circulating biomarkers of pulmonary Hypertension. J Am Coll Cardiol. 2016; 67(2): 174-189.

[11] Li M, Riddle S, Zhang H, et al. Metabolic reprogramming regulates the proliferative and inflammatory phenotype of adventitial fibroblasts in pulmonary Hypertension through the transcriptional corepressor C-terminal binding Protein-1. Circulation. 2016; 134(15): 1105-1121.

[12] Talati MH, Brittain EL, Fessel JP, et al. Mechanisms of lipid accumulation in the bone morphogenetic protein receptor type 2 mutant right ventricle. Am J Respir Crit Care Med. 2016; 194(6): 719-728.

[13] Zhao YD, Yun HZH, Peng J, et al. De novo synthesize of bile acids in pulmonary arterial hypertension lung. Metabolomics. 2014; 10(6): 1169-1175.

[14] Rhodes CJ, Ghataorhe P, Wharton J, et al. Plasma metabolomics implicates modified transfer RNAs and altered bioenergetics in the outcomes of pulmonary arterial Hypertension. Circulation. 2017; 135(5): 460-475.

[15] He YY, Yan Y, Jiang X, et al. Spermine promotes pulmonary vascular remodelling and its synthase is a therapeutic target for pulmonary arterial hypertension. Eur Respir J. 2020; 56(5): 2000522.

[16] Rhodes CJ. The cancer hypothesis of pulmonary arterial hypertension: are polyamines the new Warburg? Eur Respir J. 2020; 56(5): 2002350.

[17] He YY, Yan Y, Chen JW, et al. Plasma metabolomics in the perioperative period of defect repair in patients with pulmonary arterial hypertension associated with congenital heart disease. Acta Pharmacol Sin. 2022; 43(7): 1710-1720.

[18] Zheng HK, Zhao JH, Yan Y, et al. Metabolic reprogramming of the urea cycle pathway in experimental pulmonary arterial hypertension rats induced by monocrotaline. Respir Res. 2018; 19: 94.

[19] Zhao JH, He YY, Guo SS, et al. Circulating plasma Metabolomic profiles differentiate rodent models of pulmonary Hypertension and idiopathic pulmonary arterial Hypertension patients. Am J Hypertens. 2019; 32(11): 1109-1117.

[20] Lechartier B, Berrebeh N, Huertas A, Humbert M, Guignabert C, Tu L. Phenotypic diversity of vascular smooth muscle cells in pulmonary arterial Hypertension: implications for therapy. Chest. 2022; 161(1): 219-231.

[21] He S, Zhu T, Fang Z. The role and regulation of pulmonary artery smooth muscle cells in pulmonary Hypertension. Int J Hypertens. 2020; 2020: 1478291.

[22] Han S, Chandel NS. Lessons from cancer metabolism for pulmonary arterial Hypertension and fibrosis. Am J Respir Cell Mol Biol. 2021; 65(2): 134-145.

[23] Shi J, Yang Y, Cheng A, Xu G, He F. Metabolism of vascular smooth muscle cells in vascular diseases. Am J Physiol Heart Circ Physiol. 2020; 319(3): H613-H631.

[24] James J, Zemskova M, Eccles CA, et al. Single mutation in the NFU1 gene metabolically reprograms pulmonary artery smooth muscle cells. Arterioscler Thromb Vasc Biol. 2021; 41(2): 734-754.

[25] Ahmad I, Mui E, Galbraith L, et al. Sleeping beauty screen reveals Pparg activation in metastatic prostate cancer. Proc Natl Acad Sci USA. 2016; 113(29): 8290-8295.

[26] Wang Z, Zhang J, Ren T, Dong Z. Targeted metabolomic profiling of cardioprotective effect of Ginkgo biloba L. extract on myocardial ischemia in rats. Phytomedicine. 2016; 23(6): 621-631.

[27] Wang S, Yan Y, Xu WJ, et al. The role of glutamine and Glutaminase in pulmonary Hypertension. Front Cardiovasc Med. 2022; 9: 838657.

[28] La Frano MR, Fahrmann JF, Grapov D, et al. Umbilical cord blood metabolomics reveal distinct signatures of dyslipidemia prior to bronchopulmonary dysplasia and pulmonary hypertension. Am J Physiol Lung Cell Mol Physiol. 2018; 315(5): L870-L881.

[29] He YY, Xie XM, Zhang HD, et al. Identification of hypoxia induced metabolism associated genes in pulmonary Hypertension. Front Pharmacol. 2021; 12: 753727.

[30] Yan Y, Jiang R, Yuan P, et al. Implication of proliferation gene biomarkers in pulmonary hypertension. Animal Model Exp Med. 2021; 4(4): 369-380.

[31] Tian L, Wu D, Dasgupta A, et al. Epigenetic metabolic reprogramming of right ventricular fibroblasts in pulmonary arterial Hypertension: a pyruvate dehydrogenase kinase-dependent shift in mitochondrial metabolism promotes right ventricular fibrosis. Circ Res. 2020; 126(12): 1723-1745.

[32] Michelakis ED, Gurtu V, Webster L, et al. Inhibition of pyruvate dehydrogenase kinase improves pulmonary arterial hypertension in genetically susceptible patients. Sci Transl Med. 2017; 9: eaao4583.

[33] Caruso P, Dunmore BJ, Schlosser K, et al. 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. Circulation. 2017; 136(25): 2451-2467.

[34] Zhang H, Wang D, Li M, et al. Metabolic and proliferative state of vascular adventitial fibroblasts in pulmonary Hypertension is regulated through a MicroRNA-124/PTBP1 (Polypyrimidine tract binding protein 1)/pyruvate kinase muscle Axis. Circulation. 2017; 136(25): 2468-2485.

[35] Culley MK, Chan SY. Mitochondrial metabolism in pulmonary hypertension: beyond mountains there are mountains. J Clin Invest. 2018; 128(9): 3704-3715.

[36] Cao Y, Zhang X, Wang L, et al. PFKFB3-mediated endothelial glycolysis promotes pulmonary hypertension. Proc Natl Acad Sci U S A. 2019; 116: 13394-13403.

[37] Kovacs L, Cao Y, Han W, et al. PFKFB3 in smooth muscle promotes vascular remodeling in pulmonary arterial Hypertension. Am J Respir Crit Care Med. 2019; 200(5): 617-627.

[38] Xu JZ, Conrey A, Frey I, et al. A phase 1 dose escalation study of the pyruvate kinase activator mitapivat (AG-348) in sickle cell disease. Blood. 2022; 140(19): 2053-2062.

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