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MicroRNA-302a promotes neointimal formation following carotid artery injury in mice by targeting PHLPP2 thus increasing Akt signaling

Abstract

The excessive proliferation and migration of smooth muscle cells (SMCs) play an important role in restenosis following percutaneous coronary interventions. MicroRNAs are able to target various genes and involved in the regulation of diverse cellular processes including cell growth and proliferation. In this study we investigated whether and how MicroRNAs regulated vascular SMC proliferation and vascular remodeling following carotid artery injury in mice. We showed that carotid artery injury-induced neointimal formation was remarkably ameliorated in microRNA (miR)-302 heterozygous mice and SMC-specific miR-302 knockout mice. In contrast, delivery of miR-302a adenovirus to the injured carotid artery enhanced neointimal formation. Upregulation of miR-302a enhanced the proliferation and migration of mouse aorta SMC (MASMC) in vitro by promoting cell cycle transition, whereas miR-302a inhibition caused the opposite results. Moreover, miR-302a promoted Akt activation by corporately decreasing Akt expression and increasing Akt phosphorylation in MASMCs. Application of the Akt inhibitor GSK690693 (5 μmol/L) counteracted the functions of miR-302a in promoting MASMC proliferation and migration. We further revealed that miR-302a directly targeted at the 3′ untranslated region of PH domain and leucine rich repeat protein phosphatase 2 (PHLPP2) and negatively regulated PHLPP2 expression. Restoration of PHLPP2 abrogated the effects of miR-302a on Akt activation and MASMC motility. Furthermore, knockdown of PHLPP2 largely abolished the inhibition of neointimal formation that was observed in miR-302 heterozygous mice. Our data demonstrate that miR-302a exacerbates SMC proliferation and restenosis through increasing Akt signaling by targeting PHLPP2.

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Fig. 1: Lack of miR-302 inhibits neointimal formation induced by carotid artery injury.
Fig. 2: MiR-302a potentiates PDGF-BB-induced MASMC proliferation and migration.
Fig. 3: MiR-302a regulates MASMC proliferation dependent on Akt signaling.
Fig. 4: MiR-302a targets PHLPP2 and negatively regulates its expression.
Fig. 5: Restoration of PHLPP2 or PTEN expression attenuates the effects of miR-302a on Akt activation, MASMC proliferation, and migration.
Fig. 6: MiR-302 knockdown ameliorates neointimal formation in a PHLPP2-dependent manner.

References

  1. 1.

    Alexander MR, Owens GK. Epigenetic control of smooth muscle cell differentiation and phenotypic switching in vascular development and disease. Annu Rev Physiol. 2012;74:13–40.

    CAS  Article  Google Scholar 

  2. 2.

    Head SJ, Milojevic M, Daemen J, Ahn JM, Boersma E, Christiansen EH, et al. Mortality after coronary artery bypass grafting versus percutaneous coronary intervention with stenting for coronary artery disease: a pooled analysis of individual patient data. Lancet. 2018;391:939–48.

    Article  Google Scholar 

  3. 3.

    Fattori R, Piva T. Drug-eluting stents in vascular intervention. Lancet. 2003;361:247–9.

    Article  Google Scholar 

  4. 4.

    Xu K, Al-Ani MK, Pan X, Chi Q, Dong N, Qiu X. Plant-derived products for treatment of vascular intima hyperplasia selectively inhibit vascular smooth muscle cell functions. Evid Based Complement Altern Med. 2018;2018:3549312.

    Google Scholar 

  5. 5.

    Lv XF, Zhang YJ, Liu X, Zheng HQ, Liu CZ, Zeng XL, et al. TMEM16A ameliorates vascular remodeling by suppressing autophagy via inhibiting Bcl-2–p62 complex formation. Theranostics. 2020;10:3980–93.

    CAS  Article  Google Scholar 

  6. 6.

    Huang Y, Shen XJ, Zou Q, Wang SP, Tang SM, Zhang GZ. Biological functions of microRNAs: a review. J Physiol Biochem. 2011;67:129–39.

    CAS  Article  Google Scholar 

  7. 7.

    Liu XY, Zhang FR, Shang JY, Liu YY, Lv XF, Yuan JN, et al. Renal inhibition of miR-181a ameliorates 5-fluorouracil-induced mesangial cell apoptosis and nephrotoxicity. Cell Death Dis. 2018;9:610.

    Article  Google Scholar 

  8. 8.

    Barroso-delJesus A, Romero-Lopez C, Lucena-Aguilar G, Melen GJ, Sanchez L, Ligero G, et al. Embryonic stem cell-specific miR302-367 cluster: human gene structure and functional characterization of its core promoter. Mol Cell Biol. 2008;28:6609–19.

    CAS  Article  Google Scholar 

  9. 9.

    Barroso-del Jesus A, Lucena-Aguilar G, Menendez P. The miR-302-367 cluster as a potential stemness regulator in ESCs. Cell Cycle. 2009;8:394–8.

    CAS  Article  Google Scholar 

  10. 10.

    Parchem RJ, Moore N, Fish JL, Parchem JG, Braga TT, Shenoy A, et al. MiR-302 is required for timing of neural differentiation, neural tube closure, and embryonic viability. Cell Rep. 2015;12:760–73.

    CAS  Article  Google Scholar 

  11. 11.

    Li X, Liu LL, Yao JL, Wang K, Ai H. Human umbilical cord mesenchymal stem cell-derived extracellular vesicles inhibit endometrial cancer cell proliferation and migration through delivery of exogenous miR-302a. Stem Cells Int. 2019;2019:8108576.

    PubMed  PubMed Central  Google Scholar 

  12. 12.

    Ma YS, Lv ZW, Yu F, Chang ZY, Cong XL, Zhong XM, et al. MicroRNA-302a/d inhibits the self-renewal capability and cell cycle entry of liver cancer stem cells by targeting the E2F7/AKT axis. J Exp Clin Cancer Res. 2018;37:252.

    CAS  Article  Google Scholar 

  13. 13.

    Zhang GM, Bao CY, Wan FN, Cao DL, Qin XJ, Zhang HL, et al. MicroRNA-302a suppresses tumor cell proliferation by inhibiting AKT in prostate cancer. PLoS ONE. 2015;10:e0124410.

    Article  Google Scholar 

  14. 14.

    Li HH, Lin SL, Huang CN, Lu FJ, Chiu PY, Huang WN, et al. miR-302 attenuates amyloid-beta-induced neurotoxicity through activation of Akt signaling. J Alzheimers Dis. 2016;50:1083–98.

    CAS  Article  Google Scholar 

  15. 15.

    Stabile E, Zhou YF, Saji M, Castagna M, Shou M, Kinnaird TD, et al. Akt controls vascular smooth muscle cell proliferation in vitro and in vivo by delaying G1/S exit. Circ Res. 2003;93:1059–65.

    CAS  Article  Google Scholar 

  16. 16.

    Liang SJ, Zeng DY, Mai XY, Shang JY, Wu QQ, Yuan JN, et al. Inhibition of Orai1 store-operated calcium channel prevents foam cell formation and atherosclerosis. Arterioscler Thromb Vasc Biol. 2016;36:618–28.

    CAS  Article  Google Scholar 

  17. 17.

    Guo JW, Liu X, Zhang TT, Lin XC, Hong Y, Yu J, et al. Hepatocyte TMEM16A deletion retards NAFLD progression by ameliorating hepatic glucose metabolic disorder. Adv Sci. 2020;7:1903657.

  18. 18.

    Kessenbrock K, Plaks V, Werb Z. Matrix metalloproteinases: regulators of the tumor microenvironment. Cell. 2010;141:52–67.

    CAS  Article  Google Scholar 

  19. 19.

    Kim K, Qiang L, Hayden MS, Sparling DP, Purcell NH, Pajvani UB. mTORC1-independent Raptor prevents hepatic steatosis by stabilizing PHLPP2. Nat Commun. 2016;7:10255.

    CAS  Article  Google Scholar 

  20. 20.

    Newton AC, Trotman LC. Turning off AKT: PHLPP as a drug target. Annu Rev Pharmacol Toxicol. 2014;54:537–58.

    CAS  Article  Google Scholar 

  21. 21.

    Guo Y, Cui J, Ji Z, Cheng C, Zhang K, Zhang C, et al. miR-302/367/LATS2/YAP pathway is essential for prostate tumor-propagating cells and promotes the development of castration resistance. Oncogene. 2017;36:6336–47.

    CAS  Article  Google Scholar 

  22. 22.

    Ham O, Lee SY, Song BW, Lee CY, Lee J, Seo HH, et al. Small molecule-mediated induction of miR-9 suppressed vascular smooth muscle cell proliferation and neointima formation after balloon injury. Oncotarget. 2017;8:93360–72.

    Article  Google Scholar 

  23. 23.

    Wang DW, Wang YQ, Shu HS. MiR-16 inhibits pituitary adenoma cell proliferation via the suppression of ERK/MAPK signal pathway. Eur Rev Med Pharmacol Sci. 2018;22:1241–8.

    PubMed  Google Scholar 

  24. 24.

    Chen T, Zhou Q, Tang H, Bozkanat M, Yuan JX, Raj JU, et al. miR-17/20 Controls Prolyl hydroxylase 2 (PHD2)/hypoxia-inducible factor 1 (HIF1) to regulate pulmonary artery smooth muscle cell proliferation. J Am Heart Assoc. 2016;5:e004510.

    PubMed  PubMed Central  Google Scholar 

  25. 25.

    Liang Z, Bian X, Shim H. Inhibition of breast cancer metastasis with microRNA-302a by downregulation of CXCR4 expression. Breast Cancer Res Treat. 2014;146:535–42.

    Article  Google Scholar 

  26. 26.

    Kuppusamy KT, Sperber H, Ruohola-Baker H. MicroRNA regulation and role in stem cell maintenance, cardiac differentiation and hypertrophy. Curr Mol Med. 2013;13:757–64.

    CAS  Article  Google Scholar 

  27. 27.

    Fang YC, Yeh CH. Inhibition of miR-302 suppresses hypoxia-reoxygenation-induced H9c2 cardiomyocyte death by regulating Mcl-1 expression. Oxid Med Cell Longev. 2017;2017:7968905.

    PubMed  PubMed Central  Google Scholar 

  28. 28.

    Li G, Song Y, Li YD, Jie LJ, Wu WY, Li JZ, et al. Circulating miRNA-302 family members as potential biomarkers for the diagnosis of acute heart failure. Biomark Med. 2018;12:871–80.

    CAS  Article  Google Scholar 

  29. 29.

    Xu F, Yang J, Shang J, Lan F, Li M, Shi L, et al. MicroRNA-302d promotes the proliferation of human pluripotent stem cell-derived cardiomyocytes by inhibiting LATS2 in the Hippo pathway. Clin Sci. 2019;133:1387–99.

    CAS  Article  Google Scholar 

  30. 30.

    Shi L, Ji Y, Jiang X, Zhou L, Xu Y, Li Y, et al. Liraglutide attenuates high glucose-induced abnormal cell migration, proliferation, and apoptosis of vascular smooth muscle cells by activating the GLP-1 receptor, and inhibiting ERK1/2 and PI3K/Akt signaling pathways. Cardiovasc Diabetol. 2015;14:18.

    Article  Google Scholar 

  31. 31.

    Gao MH, Miyanohara A, Feramisco JR, Tang T. Activation of PH-domain leucine-rich protein phosphatase 2 (PHLPP2) by agonist stimulation in cardiac myocytes expressing adenylyl cyclase type 6. Biochem Biophys Res Commun. 2009;384:193–8.

    CAS  Article  Google Scholar 

  32. 32.

    Zhang J, Bai R, Li M, Ye H, Wu C, Wang C, et al. Excessive miR-25-3p maturation via N(6)-methyladenosine stimulated by cigarette smoke promotes pancreatic cancer progression. Nat Commun. 2019;10:1858.

    Article  Google Scholar 

  33. 33.

    Deng J, Ma M, Jiang W, Zhang H, Cui S. MiR-493 promotes prostate cancer cells proliferation by targeting PHLPP2 and activating Akt signaling pathway. Clin Lab. 2019;65. https://doi.org/10.7754/Clin.Lab.2018.180806.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (81525025, 81930106, 91739104, 81773723 and 81603098), National Key R&D Program of China (2017YFC0909302), Science and Technology Program of Guangzhou (201707010023), Fundamental Research Funds for the Central Universities (17ykjc29 and 17ykpy05), and High-level Health Teams of Zhuhai (2018).

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JGZ and SJL designed the research; YYL performedthe research; YYL and XL analyzed the data; SJL wrote the paper.

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Correspondence to Si-jia Liang.

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The authors declare no competing interests.

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Liu, Yy., Liu, X., Zhou, Jg. et al. MicroRNA-302a promotes neointimal formation following carotid artery injury in mice by targeting PHLPP2 thus increasing Akt signaling. Acta Pharmacol Sin 42, 550–559 (2021). https://doi.org/10.1038/s41401-020-0440-4

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Keywords

  • microRNA-302a
  • neointimal formation
  • vascular smooth muscle cell
  • proliferation
  • Akt
  • PHLPP2

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