Decreased miR‑92a‑3p expression potentially mediates the pro‑angiogenic effects of oxidative stress‑activated endothelial cell‑derived exosomes by targeting tissue factor

Li,Sufang; Yuan,Lan; Su,Lina; Lian,Zheng; Liu,Chuanfen; Zhang,Feng; Cui,Yuxia; Wu,Manyan; Chen,Hong

doi:10.3892/ijmm.2020.4713

Decreased miR‑92a‑3p expression potentially mediates the pro‑angiogenic effects of oxidative stress‑activated endothelial cell‑derived exosomes by targeting tissue factor

Authors:
- Sufang Li
- Lan Yuan
- Lina Su
- Zheng Lian
- Chuanfen Liu
- Feng Zhang
- Yuxia Cui
- Manyan Wu
- Hong Chen
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Affiliations: Department of Cardiology, Peking University People's Hospital, Beijing 100191, P.R. China, Medical and Healthy Analytical Center, Peking University, Beijing 100191, P.R. China
Published online on: August 27, 2020 https://doi.org/10.3892/ijmm.2020.4713
Pages: 1886-1898
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Angiogenesis is an essential pathological feature of vulnerable atherosclerotic plaque. Exosome‑derived microRNAs (miRNAs or miRs) have been proven to be important regulators of angiogenesis. However, the role of exosomes, which are secreted by endothelial cells (ECs) under conditions of oxidative stress, in angiogenesis remain unclear. The present study aimed to investigate the effects and mechanism of oxidative stress‑activated endothelial‑derived exosomes in angiogenesis. Exosomes were isolated from H2O2‑stimulated human umbilical vein ECs (HUVECs; termed Exo-H2O2) by differential centrifugation and characterized by transmission electron microscopy, nanoparticle tracking analysis and western blot analysis. Exo-H2O2 enhanced HUVEC proliferation, migration and tube formation, as determined by EdU incorporation assay, scratch wound migration assay and tube formation assay, respectively. miR‑92a‑3p was identified as the predominantly downregulated miRNA in the Exo-H2O2‑treated HUVECs by small RNA sequencing, and the expression of primary miR‑92a (pri‑miR‑92a‑1) was also decreased, as shown by RT‑qPCR. Similarly, the inhibition of miR‑92a‑3p promoted angiogenesis in vitro and in vivo. miR‑92a‑3p overexpression blocked the pro‑angiogenic effects of Exo-H2O2 on target ECs. Tissue factor (TF), a molecule involved in angiogenesis, was increased in HUVECs in which miR‑92a‑3p expression was downregulated, as shown by mRNA sequencing. TF was also predicted as a target of miR‑92a‑3p by using the RNAhybrid program. The overexpression or suppression of miR‑92a‑3p modified TF expression at both the mRNA and protein level, as measured by RT‑qPCR and western blot analysis, respectively. Luciferase reporter assays suggested that miR‑92a‑3p inhibited TF expression by binding to the 3' untranslated region of TF. On the whole, the findings of the present study demonstrate that exosomes released from oxidative stress‑activated ECs stimulate angiogenesis by inhibiting miR‑92a‑3p expression in recipient ECs, and TF may be involved in the regulatory effects of miR‑92a‑3p on angiogenesis.

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1	Potente M and Mäkinen T: Vascular heterogeneity and specialization in development and disease. Nat Rev Mol Cell Biol. 18:477–494. 2017. View Article : Google Scholar : PubMed/NCBI
2	Münzel T, Camici GG, Maack C, Bonetti NR, Fuster V and Kovacic JC: Impact of oxidative stress on the heart and vasculature: Part 2 of a 3-part series. J Am Coll Cardiol. 70:212–229. 2017. View Article : Google Scholar
3	Todorova D, Simoncini S, Lacroix R, Sabatier F and Dignat-George F: Extracellular vesicles in angiogenesis. Circ Res. 120:1658–1673. 2017. View Article : Google Scholar : PubMed/NCBI
4	Mathieu M, Martin-Jaular L, Lavieu G and Théry C: Specificities of secretion and uptake of exosomes and other extracellular vesicles for cell-to-cell communication. Nat Cell Biol. 21:9–17. 2019. View Article : Google Scholar : PubMed/NCBI
5	Li J, Zhang Y, Liu Y, Dai X, Li W, Cai X, Yin Y, Wang Q, Xue Y, Wang C, et al: Microvesicle-mediated transfer of microRNA-150 from monocytes to endothelial cells promotes angiogenesis. J Biol Chem. 288:23586–23596. 2013. View Article : Google Scholar : PubMed/NCBI
6	Zheng B, Yin WN, Suzuki T, Zhang XH, Zhang Y, Song LL, Jin LS, Zhan H, Zhang H, Li JS and Wen JK: Exosome-mediated miR-155 transfer from smooth muscle cells to endothelial cells induces endothelial injury and promotes atherosclerosis. Mol Ther. 25:1279–1294. 2017. View Article : Google Scholar : PubMed/NCBI
7	Deregibus MC, Cantaluppi V, Calogero R, Lo Iacono M, Tetta C, Biancone L, Bruno S, Bussolati B and Camussi G: Endothelial progenitor cell derived microvesicles activate an angiogenic program in endothelial cells by a horizontal transfer of mRNA. Blood. 110:2440–2448. 2007. View Article : Google Scholar : PubMed/NCBI
8	Gong M, Yu B, Wang J, Wang Y, Liu M, Paul C, Millard RW, Xiao DS, Ashraf M and Xu M: Mesenchymal stem cells release exosomes that transfer miRNAs to endothelial cells and promote angiogenesis. Oncotarget. 8:45200–45212. 2017. View Article : Google Scholar : PubMed/NCBI
9	Umezu T, Tadokoro H, Azuma K, Yoshizawa S, Ohyashiki K and Ohyashiki JH: Exosomal miR-135b shed from hypoxic multiple myeloma cells enhances angiogenesis by targeting factor-inhib-iting HIF-1. Blood. 124:3748–3757. 2014. View Article : Google Scholar : PubMed/NCBI
10	van Balkom BW, de Jong OG, Smits M, Brummelman J, den Ouden K, de Bree PM, van Eijndhoven MA, Pegtel DM, Stoorvogel W, Würdinger T and Verhaar MC: Endothelial cells require miR-214 to secrete exosomes that suppress senescence and induce angiogenesis in human and mouse endothelial cells. Blood. 121:3997–4006. 2013. View Article : Google Scholar : PubMed/NCBI
11	Lombardo G, Dentelli P, Togliatto G, Rosso A, Gili M, Gallo S, Deregibus MC, Camussi G and Brizzi MF: Activated Stat5 trafficking via endothelial cell-derived extracellular vesicles controls IL-3 pro-angiogenic paracrine action. Sci Rep. 6:256892016. View Article : Google Scholar : PubMed/NCBI
12	Kim YW and Byzova TV: Oxidative stress in angiogenesis and vascular disease. Blood. 123:625–631. 2014. View Article : Google Scholar :
13	Sun LL, Li WD, Lei FR and Li XQ: The regulatory role of microRNAs in angiogenesis-related diseases. J Cell Mol Med. 22:4568–4587. 2018. View Article : Google Scholar : PubMed/NCBI
14	Bonauer A, Carmona G, Iwasaki M, Mione M, Koyanagi M, Fischer A, Burchfield J, Fox H, Doebele C, Ohtani K, et al: MicroRNA-92a controls angiogenesis and functional recovery of ischemic tissues in mice. Science. 324:1710–1713. 2009. View Article : Google Scholar : PubMed/NCBI
15	Iaconetti C, Polimeni A, Sorrentino S, Sabatino J, Pironti G, Esposito G, Curcio A and Indolfi C: Inhibition of miR-92a increases endothelial proliferation and migration in vitro as well as reduces neointimal proliferation in vivo after vascular injury. Basic Res Cardiol. 107:2962012. View Article : Google Scholar : PubMed/NCBI
16	Hinkel R, Penzkofer D, Zühlke S, Fischer A, Husada W, Xu QF, Baloch E, van Rooij E, Zeiher AM, Kupatt C and Dimmeler S: Inhibition of microRNA-92a protects against ischemia/reperfusion injury in a large-animal model. Circulation. 128:1066–1075. 2013. View Article : Google Scholar : PubMed/NCBI
17	Daniel JM, Penzkofer D, Teske R, Dutzmann J, Koch A, Bielenberg W, Bonauer A, Boon RA, Fischer A, Bauersachs J, et al: Inhibition of miR-92a improves re-endothelialization and prevents neointima formation following vascular injury. Cardiovasc Res. 103:564–572. 2014. View Article : Google Scholar : PubMed/NCBI
18	Bellera N, Barba I, Rodriguez-Sinovas A, Ferret E, Asín MA, Gonzalez-Alujas MT, Pérez-Rodon J, Esteves M, Fonseca C, Toran N, et al: Single intracoronary injection of encapsulated antagomir-92a promotes angiogenesis and prevents adverse infarct remodeling. J Am Heart Assoc. 3:e0009462014. View Article : Google Scholar : PubMed/NCBI
19	Liu Y, Li Q, Hosen MR, Zietzer A, Flender A, Levermann P, Schmitz T, Frühwald D, Goody P, Nickenig G, et al: Atherosclerotic conditions promote the packaging of functional MicroRNA-92a-3p into endothelial microvesicles. Circ Res. 124:575–587. 2019. View Article : Google Scholar
20	Bang C, Batkai S, Dangwal S, Gupta SK, Foinquinos A, Holzmann A, Just A, Remke J, Zimmer K, Zeug A, et al: Cardiac fibroblast-derived microRNA passenger strand-enriched exosomes mediate cardiomyocyte hypertrophy. J Clin Invest. 124:2136–2146. 2014. View Article : Google Scholar : PubMed/NCBI
21	Li S, Chen H, Ren J, Geng Q, Song J, Lee C, Cao C, Zhang J and Xu N: MicroRNA-223 inhibits tissue factor expression in vascular endothelial cells. Atherosclerosis. 237:514–520. 2014. View Article : Google Scholar : PubMed/NCBI
22	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
23	Li S, Geng Q, Chen H, Zhang J, Cao C, Zhang F, Song J, Liu C and Liang W: The potential inhibitory effects of miR-19b on vulnerable plaque formation via the suppression of STAT3 transcriptional activity. Int J Mol Med. 41:859–867. 2018.
24	Rabiolo A, Bignami F, Rama P and Ferrari G: VesselJ: A new tool for semiautomatic measurement of corneal neovascularization. Invest Ophthalmol Vis Sci. 56:8199–8206. 2015. View Article : Google Scholar
25	van Balkom BW, Eisele AS, Pegtel DM, Bervoets S and Verhaar MC: Quantitative and qualitative analysis of small RNAs in human endothelial cells and exosomes provides insights into localized RNA processing, degradation and sorting. J Extracell Vesicles. 4:267602015. View Article : Google Scholar : PubMed/NCBI
26	Rehmsmeier M, Steffen P, Hochsmann M and Giegerich R: Fast and effective prediction of microRNA/target duplexes. RNA. 10:1507–1517. 2004. View Article : Google Scholar : PubMed/NCBI
27	Eisenreich A and Rauch U: Regulation and differential role of the tissue factor isoforms in cardiovascular biology. Trends Cardiovasc Med. 20:199–203. 2010. View Article : Google Scholar : PubMed/NCBI
28	Raposo G and Stahl PD: Extracellular vesicles: A new communication paradigm? Nat Rev Mol Cell Biol. 20:509–510. 2019. View Article : Google Scholar : PubMed/NCBI
29	Chen X, Yang F, Zhang T, Wang W, Xi W, Li Y, Zhang D, Huo Y, Zhang J, Yang A and Wang T: MiR-9 promotes tumorigenesis and angiogenesis and is activated by MYC and OCT4 in human glioma. J Exp Clin Cancer Res. 38:992019. View Article : Google Scholar : PubMed/NCBI
30	Zhuang G, Wu X, Jiang Z, Kasman I, Yao J, Guan Y, Oeh J, Modrusan Z, Bais C, Sampath D and Ferrara N: Tumour-secreted miR-9 promotes endothelial cell migration and angiogenesis by activating the JAK-STAT pathway. EMBO J. 31:3513–3523. 2012. View Article : Google Scholar : PubMed/NCBI
31	Madelaine R, Sloan SA, Huber N, Notwell JH, Leung LC, Skariah G, Halluin C, Paşca SP, Bejerano G, Krasnow MA, et al: MicroRNA-9 couples brain neurogenesis and angiogenesis. . Cell Rep. 20:1533–1542. 2017. View Article : Google Scholar : PubMed/NCBI
32	Zhang H, Qi M, Li S, Qi T, Mei H, Huang K, Zheng L and Tong Q: MicroRNA-9 targets matrix metalloproteinase 14 to inhibit invasion, metastasis, and angiogenesis of neuroblastoma cells. Mol Cancer Ther. 11:1454–1466. 2012. View Article : Google Scholar : PubMed/NCBI
33	Shyu KG, Wang BW, Pan CM, Fang WJ and Lin CM: Hyperbaric oxygen boosts long noncoding RNA MALAT1 exosome secretion to suppress microRNA-92a expression in therapeutic angiogenesis. Int J Cardiol. 274:271–278. 2019. View Article : Google Scholar
34	Eisenreich A, Bolbrinker J and Leppert U: Tissue factor: A conventional or alternative target in cancer therapy. Clin Chem. 62:563–570. 2016. View Article : Google Scholar : PubMed/NCBI
35	Versteeg HH, Schaffner F, Kerver M, Petersen HH, Ahamed J, Felding-Habermann B, Takada Y, Mueller BM and Ruf W: Inhibition of tissue factor signaling suppresses tumor growth. Blood. 111:190–199. 2008. View Article : Google Scholar
36	van den Berg YW, van den Hengel LG, Myers HR, Ayachi O, Jordanova E, Ruf W, Spek CA, Reitsma PH, Bogdanov VY and Versteeg HH: Alternatively spliced tissue factor induces angiogenesis through integrin ligation. Proc Natl Acad Sci USA. 106:19497–19502. 2009. View Article : Google Scholar : PubMed/NCBI