CircPMS1 promotes proliferation of pulmonary artery smooth muscle cells, pulmonary microvascular endothelial cells, and pericytes under hypoxia

Abstract

Background: Circular RNAs (circRNAs) have been recognized as significant regulators of pulmonary hypertension (PH); however, the differential expression and function of circRNAs in different vascular cells under hypoxia remain unknown. Here, we identified co-differentially expressed circRNAs and determined their putative roles in the proliferation of pulmonary artery smooth muscle cells (PASMCs), pulmonary microvascular endothelial cells (PMECs), and pericytes (PCs) under hypoxia.

Methods: Whole transcriptome sequencing was performed to analyze the differential expression of circRNAs in three different vascular cell types. Bioinformatic analysis was used to predict their putative biological function. Quantitative real-time polymerase chain reaction, Cell Counting Kit-8, and EdU Cell Proliferation assays were carried out to determine the role of circular postmeiotic segregation 1 (circPMS1) as well as its potential sponge mechanism in PASMCs, PMECs, and PCs.

Results: PASMCs, PMECs, and PCs exhibited 16, 99, and 31 differentially expressed circRNAs under hypoxia, respectively. CircPMS1 was upregulated in PASMCs, PMECs, and PCs under hypoxia and enhanced the proliferation of vascular cells. CircPMS1 may upregulate DEP domain containing 1 (DEPDC1) and RNA polymerase II subunit D expression by targeting microRNA-432-5p (miR-432-5p) in PASMCs, upregulate MAX interactor 1 (MXI1) expression by targeting miR-433-3p in PMECs, and upregulate zinc finger AN1-type containing 5 (ZFAND5) expression by targeting miR-3613-5p in PCs.

Conclusions: Our results suggest that circPMS1 promotes cell proliferation through the miR-432-5p/DEPDC1 or miR-432-5p/POL2D axis in PASMCs, through the miR-433-3p/MXI1 axis in PMECs, and through the miR-3613-5p/ZFAND5 axis in PCs, which provides putative targets for the early diagnosis and treatment of PH.

Keywords

circular postmeiotic segregation 1 / circular RNAs / hypoxia / pulmonary hypertension / vascular cells

References

[1] Simonneau G, Gatzoulis MA, Adatia I, et al. Updated clinical classification of pulmonary hypertension. J Am Coll Cardiol. 2013;62:D34-D41.

[2] Wang Q, Sun Y, Zhao Q, et al. Circular RNAs in pulmonary hypertension: emerging biological concepts and potential mechanism. Animal Model Exp Med. 2022;5:38-47.

[3] Ma C, Wang X, He S, et al. Ubiquitinated AIF is a major mediator of hypoxia-induced mitochondrial dysfunction and pulmonary artery smooth muscle cell proliferation. Cell Biosci. 2022;12:9.

[4] Kurakula K, Smolders V, Tura-Ceide O, Jukema JW, Quax PHA, Goumans MJ. Endothelial dysfunction in pulmonary hypertension: cause or consequence? Biomedicine. 2021;9:57.

[5] Ricard N, Tu L, Le Hiress M, et al. Increased pericyte coverage mediated by endothelial-derived fibroblast growth factor-2 and interleukin-6 is a source of smooth muscle-like cells in pulmonary hypertension. Circulation. 2014;129:1586-1597.

[6] Memczak S, Jens M, Elefsinioti A, et al. Circular RNAs are a large class of animal RNAs with regulatory potency. Nature. 2013;495:333-338.

[7] Kristensen LS, Andersen MS, Stagsted LVW, Ebbesen KK, Hansen TB, Kjems J. The biogenesis, biology and characterization of circular RNAs. Nat Rev Genet. 2019;20:675-691.

[8] Geng Y, Jiang J, Wu C. Function and clinical significance of circRNAs in solid tumors. J Hematol Oncol. 2018;11:98.

[9] Zhang J, Li Y, Qi J, et al. Circ-calm4 serves as an miR-337-3p sponge to regulate Myo10 (myosin 10) and promote pulmonary artery smooth muscle proliferation. Hypertension. 2020;75:668-679.

[10] Jiang Y, Liu H, Yu H, et al. Circular RNA Calm4 regulates hypoxia-induced pulmonary arterial smooth muscle cells pyroptosis via the circ-Calm4/miR-124-3p/PDCD6 Axis. Arterioscler Thromb Vasc Biol. 2021;41:1675-1693.

[11] Yuan P, Wu WH, Gong SG, et al. Impact of circGSAP in peripheral blood mononuclear cells on idiopathic pulmonary arterial hypertension. Am J Respir Crit Care Med. 2021;203:1579-1583.

[12] Sun Y, Wu W, Zhao Q, et al. CircGSAP regulates the cell cycle of pulmonary microvascular endothelial cells via the miR-942-5p sponge in pulmonary hypertension. Front Cell Dev Biol. 2022;10:967708.

[13] Sun Y, Jiang R, Hu X, et al. CircGSAP alleviates pulmonary microvascular endothelial cells dysfunction in pulmonary hypertension via regulating miR-27a-3p/BMPR2 axis. Respir Res. 2022;23:322.

[14] Dang RY, Liu FL, Li Y. Circular RNA hsa_circ_0010729 regulates vascular endothelial cell proliferation and apoptosis by targeting the miR-186/HIF-1alpha axis. Biochem Biophys Res Commun. 2017;490:104-110.

[15] Zhou S, Jiang H, Li M, et al. Circular RNA hsa_circ_0016070 is associated with pulmonary arterial hypertension by promoting PASMC proliferation. Mol Ther Nucleic Acids. 2019;18:275-284.

[16] Jiang Q, Liu C, Li CP, et al. Circular RNA-ZNF532 regulates diabetes-induced retinal pericyte degeneration and vascular dysfunction. J Clin Invest. 2020;130:3833-3847.

[17] Lv Z, Jiang R, Hu X, et al. Dysregulated lncRNA TUG1 in different pulmonary artery cells under hypoxia. Ann Transl Med. 2021;9:879.

[18] Gao D, Qi X, Zhang X, Fang K, Guo Z, Li L. hsa_circRNA_0006528 as a competing endogenous RNA promotes human breast cancer progression by sponging miR-7-5p and activating the MAPK/ERK signaling pathway. Mol Carcinog. 2019;58:554-564.

[19] Zheng SQ, Qi Y, Wu J, et al. CircPCMTD1 acts as the sponge of miR-224-5p to promote glioma progression. Front Oncol. 2019;9:398.

[20] Zhang H, Wang X, Huang H, Wang Y, Zhang F, Wang S. Hsa_circ_0067997 promotes the progression of gastric cancer by inhibition of miR-515-5p and activation of X chromosome-linked inhibitor of apoptosis (XIAP). Artif Cells Nanomed Biotechnol. 2019;47:308-318.

[21] Hang D, Zhou J, Qin N, et al. A novel plasma circular RNA circFARSA is a potential biomarker for non-small cell lung cancer. Cancer Med. 2018;7:2783-2791.

[22] Jiricny J. Postreplicative mismatch repair. Cold Spring Harb Perspect Biol. 2013;5:a012633.

[23] O’Brien V, Brown R. Signalling cell cycle arrest and cell death through the MMR system. Carcinogenesis. 2006;27:682-692.

[24] Liu JY, Qian CY, Gao YF, Chen J, Zhou HH, Yin JY. Association between DNA mismatch repair gene polymorphisms and platinum-based chemotherapy toxicity in non-small cell lung cancer patients. Chin J Cancer. 2017;36:12.

[25] Vymetalkova V, Pardini B, Rosa F, et al. Variations in mismatch repair genes and colorectal cancer risk and clinical outcome. Mutagenesis. 2014;29:259-265.

[26] Erdeniz N, Dudley S, Gealy R, Jinks-Robertson S, Liskay RM. Novel PMS1 alleles preferentially affect the repair of primer strand loops during DNA replication. Mol Cell Biol. 2005;25:9221-9231.

[27] Wang Y, Zhou X, Song Y, et al. The mismatch repair gene hPMS1 (human postmeiotic segregation1) is down regulated in oral squamous cell carcinoma. Gene. 2013;524:28-34.

[28] Dong J, Hu Z, Shu Y, et al. Potentially functional polymorphisms in DNA repair genes and non-small-cell lung cancer survival: a pathway-based analysis. Mol Carcinog. 2012;51:546-552.

[29] Yang J, Huang Y, Feng Y, et al. Associations of genetic variations in mismatch repair genes MSH3 and PMS1 with acute adverse events and survival in patients with rectal cancer receiving postoperative chemoradiotherapy. Cancer Res Treat. 2019;51:1198-1206.

[30] Wu S, Chen L, Zhou X. Circular RNAs in the regulation of cardiac hypertrophy. Mol Ther Nucleic Acids. 2022;27:484-490.

[31] Bella ED, Koch J, Baerenfaller K. Translation and emerging functions of non-coding RNAs in inflammation and immunity. Allergy. 2022;77:2025-2037.

[32] Liu J, Feng G, Li Z, Li R, Xia P. Knockdown of CircCRIM1 inhibits HDAC4 to impede osteosarcoma proliferation, migration, and invasion and facilitate autophagy by targeting miR-432-5p. Cancer Manag Res. 2020;12:10199-10210.

[33] Jin M, Zhang F, Li Q, Xu R, Liu Y, Zhang Y. Circ_0011292 knockdown mitigates progression and drug resistance in PTX-resistant non-small-cell lung cancer cells by regulating miR-433-3p/CHEK1 axis. Thorac Cancer. 2022;13:1276-1288.

[34] Cao R, Wang K, Long M, et al. miR-3613-5p enhances the metastasis of pancreatic cancer by targeting CDK6. Cell Cycle. 2020;19:3086-3095.

[35] Caruso P, Dempsie Y, Stevens HC, et al. A role for miR-145 in pulmonary arterial hypertension: evidence from mouse models and patient samples. Circ Res. 2012;111:290-300.

[36] McLendon JM, Joshi SR, Sparks J, et al. Lipid nanoparticle delivery of a microRNA-145 inhibitor improves experimental pulmonary hypertension. J Control Release. 2015;210:67-75.

[37] Yue Y, Zhang Z, Zhang L, Chen S, Guo Y, Hong Y. miR-143 and miR-145 promote hypoxia-induced proliferation and migration of pulmonary arterial smooth muscle cells through regulating ABCA1 expression. Cardiovasc Pathol. 2018;37:15-25.

[38] Valadi H, Ekstrom K, Bossios A, Sjostrand M, Lee JJ, Lotvall JO. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol. 2007;9:654-659.

[39] Deng L, Blanco FJ, Stevens H, et al. MicroRNA-143 activation regulates smooth muscle and endothelial cell crosstalk in pulmonary arterial hypertension. Circ Res. 2015;117:870-883.

[40] Shen J, Xi M. DEPDC1 is highly expressed in lung adenocarcinoma and promotes tumor cell proliferation. Zhongguo Fei Ai Za Zhi. 2021;24:453-460.

[41] Li Y, Tian Y, Zhong W, et al. Artemisia argyi essential oil inhibits hepatocellular carcinoma metastasis via suppression of DEPDC1 dependent Wnt/beta-catenin signaling pathway. Front Cell Dev Biol. 2021;9:664791.

[42] Maeta M, Kataoka M, Nishiya Y, Ogino K, Kashima M, Hirata H. RNA polymerase II subunit D is essential for zebrafish development. Sci Rep. 2020;10:13213.

[43] Yamada Y, Nishikawa R, Kato M, et al. Regulation of HMGB3 by antitumor miR-205-5p inhibits cancer cell aggressiveness and is involved in prostate cancer pathogenesis. J Hum Genet. 2018;63:195-205.

[44] Maiuthed A, Prakhongcheep O, Chanvorachote P. Microarray-based analysis of genes, transcription factors, and epigenetic modifications in lung cancer exposed to nitric oxide. Cancer Genomics Proteomics. 2020;17:401-415.

[45] Dong L, Liu X, Wu B, et al. Mxi1-0 promotes hypoxic pulmonary hypertension via ERK/c-Myc-dependent proliferation of arterial smooth muscle cells. Front Genet. 2022;13:810157.

[46] Liu P, Wang Y, Duan L. ZFAND5 is an independent prognostic biomarker of perihilar cholangiocarcinoma. Front Oncol. 2022;12:955670.

RIGHTS & PERMISSIONS

2023 The Author(s). Animal Models and Experimental Medicine published by John Wiley & Sons Australia, Ltd on behalf of The Chinese Association for Laboratory Animal Sciences.