WWC3 inhibits intimal proliferation following vascular injury via the Hippo signaling pathway

Chen,Beijia; Liu,Guinan

doi:10.3892/mmr.2018.8484

WWC3 inhibits intimal proliferation following vascular injury via the Hippo signaling pathway

Authors:
- Beijia Chen
- Guinan Liu
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Affiliations: Department of Cardiology, The First Affiliated Hospital of The China Medical University, Shenyang, Liaoning 110001, P.R. China
Published online on: January 25, 2018 https://doi.org/10.3892/mmr.2018.8484
Pages: 5175-5183
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The Hippo signaling pathway is involved in the formation and development of the cardiovascular system. In the present study, the effects of WWC family member 3 (WWC3) on vascular smooth muscle cells (VSMCs) following injury were investigated, in addition to the associated mechanisms underlying this process. Platelet‑derived growth factor BB (PDGF‑BB) was used as a cell injury factor, and rats with balloon injuries were used as a model of carotid intimal injury. Furthermore, the expression levels of WWC3 in VSMCs and arteries post‑injury were investigated, in addition to the effect of WWC3 on the proliferation and migration of VSMCs. The results demonstrated that following injury, WWC3 expression was suppressed in VSMCs and the rat carotid artery, and the activity of the Hippo signaling pathway was significantly downregulated. In addition, the expression of YY1‑associated protein‑1 (YAP) and a number of its downstream target genes, including connective tissue growth factor (CTGF), were enhanced, thus enhancing the proliferation and migration of VSMCs. Knockdown of WWC3 suppressed the levels of large tumor suppressor kinase 1 (LATS1) expression and YAP phosphorylation, and the expression of YAP, CTGF and cyclin E was subsequently enhanced, thus promoting cell proliferation and migration. Similar results were obtained following overexpression of WWC3. Treatment with PDGF‑BB was revealed to suppress the proliferation and migration of VSMCs transfected with the WWC3 plasmid, compared with VSMCs transfected with an empty vector. The present study demonstrated that WWC3 may interact with LATS1 in order to upregulate the Hippo signaling pathway via co‑immunoprecipitation and enhancement of the phosphorylation of LATS1, in addition to the corresponding suppression of the nuclear import of YAP. However, VSMCs transfected with WWC3 plasmid with a deletion of the WW domain fail to exhibit this effect. These results suggested that WWC3 expression is downregulated in VSMCs during neointimal hyperplasia following injury (PDGF‑BB stimulation or balloon injury). WWC3 upregulates the activity of the Hippo signaling pathway, and weakens the proliferation and migration of VSMCs. Furthermore, the results of the present study suggested that WWC3 may interact with LATS1 to promote the phosphorylation of YAP and reduce its nuclear translocation, upregulate the activity of the Hippo pathway, and suppress the proliferation and migration of VSMCs following injury.

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1	Osherov AB, Gotha L, Cheema AN, Qiang BP and Strauss BH: Proteins mediating collagen biosynthesis and accumulation in arterial repair: Novel targets for anti-restenosis therapy. Cardiovasc Res. 91:16–26. 2011. View Article : Google Scholar : PubMed/NCBI
2	Rodriguez-Menocal L, St-Pierre M, Wei Y, Khan S, Mateu D, Calfa M, Rahnemai-Azar AA, Striker G, Pham SM and Vazquez-Padron RI: The origin of post-injury neointimal cells in the rat balloon injury model. Cardiovasc Res. 81:46–53. 2009. View Article : Google Scholar : PubMed/NCBI
3	Zhang L, Yang S, Wennmann DO, Chen Y, Kremerskothen J and Dong J: KIBRA: In the brain and beyond. Cell Signal. 26:1392–1399. 2014. View Article : Google Scholar : PubMed/NCBI
4	Buttitta LA and Edgar BA: How size is controlled: From hippos to yorkies. Nat Cell Biol. 9:1225–1227. 2007. View Article : Google Scholar : PubMed/NCBI
5	Pan D: Hippo signaling in organ size control. Genes Dev. 21:886–897. 2007. View Article : Google Scholar : PubMed/NCBI
6	Mo JS, Park HW and Guan KL: The Hippo signaling pathway in stem cell biology and cancer. EMBO Rep. 15:642–656. 2014.PubMed/NCBI
7	Dong J, Feldmann G, Huang J, Wu S, Zhang N, Comerford SA, Gayyed MF, Anders RA, Maitra A and Pan D: Elucidation of a universal size control mechanism in Drosophila and mammals. Cell. 130:1120–1133. 2007. View Article : Google Scholar : PubMed/NCBI
8	Harvey KF, Zhang X and Thomas DM: The Hippo pathway and human cancer. Nat Rev Cancer. 13:246–257. 2013. View Article : Google Scholar : PubMed/NCBI
9	Han Q, Lin X, Zhang X, Jiang G, Zhang Y, Miao Y, Rong X, Zheng X, Han Y, Han X, et al: WWC3 regulates the Wnt and Hippo pathways via Dishevelled proteins and large tumour suppressor 1, to suppress lung cancer invasion and metastasis. J Pathol. 242:435–447. 2017. View Article : Google Scholar : PubMed/NCBI
10	Doherty TM, Shah PK and Rajavashisth TB: Cellular origins of atherosclerosis: Towards ontogenetic endgame? FASEB J. 17:592–597. 2003. View Article : Google Scholar : PubMed/NCBI
11	Heallen T, Zhang M, Wang J, Bonilla-Claudio M, Klysik E, Johnson RL and Martin JF: Hippo pathway inhibits Wnt signaling to restrain cardiomyocyte proliferation and heart size. Science. 332:458–461. 2011. View Article : Google Scholar : PubMed/NCBI
12	Wang X, Hu G, Gao X, Wang Y, Zhang W, Harmon EY, Zhi X, Xu Z, Lennartz MR, Barroso M, et al: The induction of yes-associated protein expression after arterial injury is crucial for smooth muscle phenotypic modulation and neointima formation. Arterioscler Thromb Vasc Biol. 32:2662–2669. 2012. View Article : Google Scholar : PubMed/NCBI
13	Xie C, Guo Y, Zhu T, Zhang J, Ma PX and Chen YE: Yap1 protein regulates vascular smooth muscle cell phenotypic switch by interaction with myocardin. J Biol Chem. 287:14598–14605. 2012. View Article : Google Scholar : PubMed/NCBI
14	Genevet A, Wehr MC, Brain R, Thompson BJ and Tapon N: Kibra is a regulator of the Salvador/Warts/Hippo signaling network. Dev Cell. 18:300–308. 2010. View Article : Google Scholar : PubMed/NCBI
15	Xiao L, Chen Y, Ji M, Volle DJ, Lewis RE, Tsai MY and Dong J: KIBRA protein phosphorylation is regulated by mitotic kinase aurora and protein phosphatase 1. J Biol Chem. 286:36304–36315. 2011. View Article : Google Scholar : PubMed/NCBI
16	Wennmann DO, Schmitz J, Wehr MC, Krahn MP, Koschmal N, Gromnitza S, Schulze U, Weide T, Chekuri A, Skryabin BV, et al: Evolutionary and molecular facts link the WWC protein family to hippo signaling. Mol Biol Evol. 31:1710–1723. 2014. View Article : Google Scholar : PubMed/NCBI
17	Wang TR, Yang G and Liu GN: DNA enzyme ED5 depletes egr-1 and inhibits neointimal hyperplasia in rats. Cardiology. 125:192–200. 2013. View Article : Google Scholar : PubMed/NCBI
18	Zhang W, Gao Y, Li P, Shi Z, Guo T, Li F, Han X, Feng Y, Zheng C, Wang Z, et al: VGLL4 functions as a new tumor suppressor in lung cancer by negatively regulating the YAP-TEAD transcriptional complex. Cell Res. 24:331–343. 2014. View Article : Google Scholar : PubMed/NCBI
19	Morin-Kensicki EM, Boone BN, Howell M, Stonebraker JR, Teed J, Alb JG, Magnuson TR, O'Neal W and Milgram SL: Defects in yolk sac vasculogenesis, chorioallantoic fusion, and embryonic axis elongation in mice with targeted disruption of yap65. Mol Cell Biol. 26:77–87. 2006. View Article : Google Scholar : PubMed/NCBI
20	von Gise A, Lin Z, Schlegelmilch K, Honor LB, Pan GM, Buck JN, Ma Q, Ishiwata T, Zhou B, Camargo FD and Pu WT: YAP1, the nuclear target of Hippo signaling, stimulates heart growth through cardiomyocyte proliferation but not hypertrophy. Proc Natl Acad Sci USA. 109:pp. 2394–2399. 2012; View Article : Google Scholar : PubMed/NCBI
21	Yamamoto S, Yang G, Zablocki D, Liu J, Hong C, Kim SJ, Soler S, Odashima M, Thaisz J, Yehia G, et al: Activation of mst1 causes dilated cardiomyopathy by stimulating apoptosis without compensatory ventricular myocyte hypertrophy. J Clin Invest. 111:1463–1474. 2003. View Article : Google Scholar : PubMed/NCBI
22	Odashima M, Usui S, Takagi H, Hong C, Liu J, Yokota M and Sadoshima J: Inhibition of endogenous mst1 prevents apoptosis and cardiac dysfunction without affecting cardiac hypertrophy after myocardial infarction. Circ Res. 100:1344–1352. 2007. View Article : Google Scholar : PubMed/NCBI
23	Ono H, Ichiki T, Ohtsubo H, Fukuyama K, Imayama I, Hashiguchi Y, Sadoshima J and Sunagawa K: Critical role of Mst1 in vascular remodeling after injury. Arterioscler Thromb Vasc Biol. 25:1871–1876. 2005. View Article : Google Scholar : PubMed/NCBI
24	Liu X, Cheng Y, Chen X, Yang J, Xu L and Zhang C: MicroRNA-31 regulated by the extracellular regulated kinase is involved in vascular smooth muscle cell growth via large tumor suppressor homolog 2. J Biol Chem. 286:42371–42380. 2011. View Article : Google Scholar : PubMed/NCBI
25	Sudol M: Yes-associated protein (YAP65) is a proline-rich phosphoprotein that binds to the SH3 domain of the Yes proto-oncogene product. Oncogene. 9:2145–2152. 1994.PubMed/NCBI
26	Wang Y, Hu G, Liu F, Wang X, Wu M, Schwarz J and Zhou J: Deletion of yes-associated protein (YAP) specifically in cardiac and vascular smooth muscle cells reveals a crucial role for YAP in mouse cardiovascular development. Circ Res. 114:957–965. 2014. View Article : Google Scholar : PubMed/NCBI
27	Zhao B, Lei QY and Guan KL: The Hippo-YAP pathway: New connections between regulation of organ size and cancer. Curr Opin Cell Biol. 20:638–646. 2008. View Article : Google Scholar : PubMed/NCBI
28	Zhao B, Wei X, Li W, Udan RS, Yang Q, Kim J, Xie J, Ikenoue T, Yu J, Li L, et al: Inactivation of YAP oncoprotein by the Hippo pathway is involved in cell contact inhibition and tissue growth control. Genes Dev. 21:2747–2761. 2007. View Article : Google Scholar : PubMed/NCBI
29	Zhao B, Tumaneng K and Guan KL: The Hippo pathway in organ size control, tissue regeneration and stem cell self-renewal. Nat Cell Biol. 13:877–883. 2011. View Article : Google Scholar : PubMed/NCBI
30	Kudryashova TV, Goncharov DA, Pena A, Kelly N, Vanderpool R, Baust J, Kobir A, Shufesky W, Mora AL, Morelli AE, et al: HIPPO-integrin linked kinase cross-talk controls self-sustaining proliferation and survival in pulmonary hypertension. Am J Respir Crit Care Med. 194:866–877. 2016. View Article : Google Scholar : PubMed/NCBI