Lin28b delays vasculature aging by reducing platelet-derived growth factor-beta resistance in senescent vascular smooth muscle cells


      • PDGFB resistance happens in aging vascular and senescent VSMCs, and may be due to the declined expression of PDGFB and PDGFRB.
      • Lin28b decreases while the downstream let-7 family increases with aging.
      • Let-7 enhances VSMCs resistance to PDGFB and induces VSMCs senescence by downregulating PDGFB and PDGFRB.
      • Overexpression of Lin28b reverses PDGFB resistance and delays senescence in VSMCs, which can be blocked by let-7.


      Background and aims

      Platelet-derived growth factor-β (PDGFB) is an important mediator of vascular smooth muscle cell (VSMC) proliferation, and PDGFB resistance is observed in senescent VSMCs. Lin28b is a stemness regulator in the embryo; however, its role in vasculature aging and VSMC senescence is unknown. We aimed to investigate whether Lin28b could restore the VSMC response to PDGFB and delay vasculature aging.


      ApoE−/− mice were fed a high-fat diet for different weeks to establish an aging model. PDGFB resistance was observed using EdU staining in vessel culture in vitro. Quantitative polymerase chain reaction and in situ hybridization were used to detect let-7 expression. Senescence was identified by Western blotting, senescence-associated beta-galactosidase activity or Sudan Black B staining, and VSMC function was determined using CCK-8, migration, and enzyme-linked immunosorbent assays.


      Vessels from aged mice showed poor responses to PDGFB stimulation compared with those from young mice; similar results were found in senescent VSMCs. The expression levels of Lin28b and PDGF receptor-β were downregulated in aging vasculature and senescent VSMCs, whereas let-7 family levels increased with aging and VSMC passage growth. Transfection of VSMCs with let-7c induced PDGFB resistance and accelerated VSMC senescence, whereas blocking let-7c restored PDGFB reactions in VSMCs. Overexpression of Lin28b protein by lentivirus resulted in the restoration of PDGFB reactions and delayed VSMC senescence, which was blocked by a let-7c mimic.


      This study reveals the role of Lin28b in delaying vasculature aging by decreasing senescent VSMC PDGFB resistance mediated by let-7.

      Graphical abstract


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        • Bao M.H.
        • Zhang Y.W.
        • Lou X.Y.
        • Cheng Y.
        • Zhou H.H.
        Protective effects of let-7a and let-7b on oxidized low-density lipoprotein induced endothelial cell injuries.
        PLoS One. 2014; 9e106540
        • Basatemur G.L.
        • Jorgensen H.F.
        • Clarke M.C.H.
        • Bennett M.R.
        • Mallat Z.
        Vascular smooth muscle cells in atherosclerosis.
        Nat. Rev. Cardiol. 2019; 16: 727-744
        • Bernstein D.L.
        • Jiang X.
        • Rom S.
        let-7 microRNAs: their role in cerebral and cardiovascular diseases, inflammation, cancer, and their regulation.
        Biomedicines. 2021; 9
        • Chen P.Y.
        • Qin L.
        • Barnes C.
        • Charisse K.
        • Yi T.
        • et al.
        FGF regulates TGF-beta signaling and endothelial-to-mesenchymal transition via control of let-7 miRNA expression.
        Cell Rep. 2012; 2: 1684-1696
        • Chi C.
        • Li D.J.
        • Jiang Y.J.
        • Tong J.
        • Fu H.
        • et al.
        Vascular smooth muscle cell senescence and age-related diseases: state of the art.
        Biochim. Biophys. Acta, Mol. Basis Dis. 2019; 1865: 1810-1821
        • Chun-Hsu Pan C.-J.C.
        • Shih Chun-Ming
        • Wang Ming-Fu
        • Wang Jie-Yu
        • Wu Chieh-Hsi
        Oxidative stress-induced cellular senescence desensitizes cell growth and migration of vascular smooth muscle cells through down-regulation of platelet-derived growth factor receptor-beta.
        Aging. 2019; 11: 8085-8102
        • Coleman Jr., L.G.
        • Zou J.
        • Crews F.T.
        Microglial-derived miRNA let-7 and HMGB1 contribute to ethanol-induced neurotoxicity via TLR7.
        J. Neuroinflammation. 2017; 14: 22
        • Di Micco R.
        • Krizhanovsky V.
        • Baker D.
        • d'Adda di Fagagna F.
        Cellular senescence in ageing: from mechanisms to therapeutic opportunities.
        Nat. Rev. Mol. Cell Biol. 2021; 22: 75-95
        • Ding Y.N.
        • Tang X.
        • Chen H.Z.
        • Liu D.P.
        Epigenetic regulation of vascular aging and age-related vascular diseases.
        Adv. Exp. Med. Biol. 2018; 1086: 55-75
        • Ding Y.N.
        • Wang H.Y.
        • Chen H.Z.
        • Liu D.P.
        Targeting senescent cells for vascular aging and related diseases.
        J. Mol. Cell. Cardiol. 2021; 162: 43-52
        • Dutta S.
        • Sengupta P.
        Men and mice: relating their ages.
        Life Sci. 2016; 152: 244-248
        • Eoin Brennan B.W.
        • McClelland Aaron
        • Mohan Muthukumar
        • Marai Mariam
        • Beuscart Ophelie
        • Derouiche Sinda
        • Gray Stephen
        • Pickering Raelene
        • Tikellis Chris
        • de Gaetano Monica
        • Barry Mary
        • Belton Orina
        • Ali-Shah Syed Tasadaque
        • Guiry Patrick
        • Jandeleit-Dahm Karin A.M.
        • Cooper Mark E.
        Catherine godson, phillip kantharidis, protective effect of let-7 miRNA family in regulating inflammation in diabetes-associated atherosclerosis.
        Diabetes. 2017; 66: 2266-2277
        • Evangelou K.
        • Gorgoulis V.G.
        Sudan black B, the specific histochemical stain for lipofuscin: a novel method to detect senescent cells.
        Methods Mol. Biol. 2017; 1534: 111-119
        • Evangelou K.
        • Lougiakis N.
        • Rizou S.V.
        • Kotsinas A.
        • Kletsas D.
        • et al.
        Robust, universal biomarker assay to detect senescent cells in biological specimens.
        Aging Cell. 2017; 16: 192-197
        • Gerthoffer W.T.
        Mechanisms of vascular smooth muscle cell migration.
        Circ. Res. 2007; 100: 607-621
        • Gomez D.
        • Owens G.K.
        Smooth muscle cell phenotypic switching in atherosclerosis.
        Cardiovasc. Res. 2012; 95: 156-164
        • Grootaert M.O.J.
        • Moulis M.
        • Roth L.
        • Martinet W.
        • Vindis C.
        • et al.
        Vascular smooth muscle cell death, autophagy and senescence in atherosclerosis.
        Cardiovasc. Res. 2018; 114: 622-634
        • Jay M Edelberg S.H.L.
        • Kaur Manmeen
        • Tang Lilong
        • Feirt Nikki M.
        • McCabe Samuel
        • Bramwell Orville
        • Wong S Chiu
        • Hong Mun K.
        Platelet-derived growth factor-AB limits the extent of myocardial infarction in a rat model: feasibility of restoring impaired angiogenic capacity in the aging heart.
        Circulation. 2002; 105: 608-613
        • Jay D.C.
        • Edelberg M.
        Munira xaymardan, translation of PDGF cardioprotective pathways.
        Cardiovasc. Toxicol. 2003; (03): 27-35
        • Jiang S.
        • Baltimore D.
        RNA-binding protein Lin28 in cancer and immunity.
        Cancer Lett. 2016; 375: 108-113
        • Johnson A.A.
        • Stolzing A.
        The role of lipid metabolism in aging, lifespan regulation, and age-related disease.
        Aging Cell. 2019; 18e13048
        • Jun-Hao E.T.
        • Gupta R.R.
        • Shyh-Chang N.
        Lin28 and let-7 in the metabolic physiology of aging.
        Trends Endocrinol. Metabol. 2016; 27: 132-141
        • Kim J.H.
        • Kim H.S.
        Downregulation of angiotensin II-induced 12-lipoxygenase expression and cell proliferation in vascular smooth muscle cells from spontaneously hypertensive rats by CCL5.
        KOREAN J. PHYSIOL. PHARMACOL. 2009; 13: 385-392
        • Lu Q.B.
        • Wan M.Y.
        • Wang P.Y.
        • Zhang C.X.
        • Xu D.Y.
        • et al.
        Chicoric acid prevents PDGF-BB-induced VSMC dedifferentiation, proliferation and migration by suppressing ROS/NFkappaB/mTOR/P70S6K signaling cascade.
        Redox Biol. 2018; 14: 656-668
        • Bennett G.I.E.M.R.
        • Schwartz S.M.
        Apoptosis of human vascular smooth muscle cells derived from normal vessels and coronary atherosclerotic plaques.
        Comparative Study. 1995; 95
        • Markopoulos G.S.
        • Roupakia E.
        • Tokamani M.
        • Vartholomatos G.
        • Tzavaras T.
        • et al.
        Senescence-associated microRNAs target cell cycle regulatory genes in normal human lung fibroblasts.
        Exp. Gerontol. 2017; 96: 110-122
        • Minamino T.
        Role of cellular senescence in lifestyle-related disease.
        Circ. J. 2010; 74: 2527-2533
        • Newman A.A.C.
        • Serbulea V.
        • Baylis R.A.
        • Shankman L.S.
        • Bradley X.
        • et al.
        Multiple cell types contribute to the atherosclerotic lesion fibrous cap by PDGFRbeta and bioenergetic mechanisms.
        Nat Metab. 2021; 3: 166-181
        • Noureddine H.
        • Gary-Bobo G.
        • Alifano M.
        • Marcos E.
        • Saker M.
        • et al.
        Pulmonary artery smooth muscle cell senescence is a pathogenic mechanism for pulmonary hypertension in chronic lung disease.
        Circ. Res. 2011; 109: 543-553
        • Ovadya Y.
        • Krizhanovsky V.
        Strategies targeting cellular senescence.
        J. Clin. Invest. 2018; 128: 1247-1254
        • Paredes F.
        • Williams H.C.
        • Quintana R.A.
        • San Martin A.
        Mitochondrial protein Poldip2 (polymerase delta interacting protein 2) controls vascular smooth muscle differentiated phenotype by O-linked GlcNAc (N-acetylglucosamine) transferase-dependent inhibition of a ubiquitin proteasome system.
        Circ. Res. 2020; 126: 41-56
        • Partridge L.
        • Deelen J.
        • Slagboom P.E.
        Facing up to the global challenges of ageing.
        Nature. 2018; 561: 45-56
        • Perinthottathil Sreejith W.J.
        Van To, Yong Hun Jo, Benoit Biteau, Changsoo Kim Lin28 is a critical factor in the function and aging of Drosophila testis stem cell niche.
        Aging (Albany NY). 2019; 11: 855-873
        • Qin B.
        • Xiao B.
        • Liang D.
        • Li Y.
        • Jiang T.
        • et al.
        MicroRNA let-7c inhibits Bcl-xl expression and regulates ox-LDL-induced endothelial apoptosis.
        BMB Rep. 2012; 45: 464-469
        • Rea I.M.
        • Gibson D.S.
        • McGilligan V.
        • McNerlan S.E.
        • Alexander H.D.
        • et al.
        Age and age-related diseases: role of inflammation triggers and cytokines.
        Front. Immunol. 2018; 9: 586
        • Shyh-Chang N.
        • Daley G.Q.
        Lin28: primal regulator of growth and metabolism in stem cells.
        Cell Stem Cell. 2013; 12: 395-406
        • Thornton J.E.
        • Gregory R.I.
        How does Lin28 let-7 control development and disease?.
        Trends Cell Biol. 2012; 22: 474-482
        • Wang D.
        • Hou L.
        • Nakamura S.
        • Su M.
        • Li F.
        • et al.
        LIN-28 balances longevity and germline stem cell number in Caenorhabditis elegans through let-7/AKT/DAF-16 axis.
        Aging Cell. 2017; 16: 113-124
        • Wang S.
        • Lai X.
        • Deng Y.
        • Song Y.
        Correlation between mouse age and human age in anti-tumor research: significance and method establishment.
        Life Sci. 2020; 242117242
        • Weng J.
        • Wang C.
        • Zhong W.
        • Li B.
        • Wang Z.
        • et al.
        Activation of CD137 signaling promotes angiogenesis in atherosclerosis via modulating endothelial smad1/5-NFATc1 pathway.
        J. Am. Heart Assoc. 2017; 6
        • Xaymardan M.
        • Zheng J.
        • Duignan I.
        • Chin A.
        • Holm J.M.
        • et al.
        Senescent impairment in synergistic cytokine pathways that provide rapid cardioprotection in the rat heart.
        J. Exp. Med. 2004; 199: 797-804
        • Xu F.
        • Pang L.
        • Cai X.
        • Liu X.
        • Yuan S.
        • et al.
        let-7-repressesed Shc translation delays replicative senescence.
        Aging Cell. 2014; 13: 185-192
        • Zaidi M.
        • Lizneva D.
        • Yuen T.
        The role of PDGF-BB in the bone-vascular relationship during aging.
        J. Clin. Invest. 2021; 131
        • Zhong W.
        • Li B.
        • Xu Y.
        • Yang P.
        • Chen R.
        • et al.
        Hypermethylation of the micro-RNA 145 promoter is the key regulator for NLRP3 inflammasome-induced activation and plaque formation.
        JACC Basic Transl Sci. 2018; 3: 604-624
        • Zhong W.
        • Li B.
        • Yang P.
        • Chen R.
        • Wang C.
        • et al.
        CD137-CD137L interaction modulates neointima formation and the phenotype transformation of vascular smooth muscle cells via NFATc1 signaling.
        Mol. Cell. Biochem. 2018; 439: 65-74
        • Zhou J.L.
        • Deng S.
        • Fang H.S.
        • Yu G.
        • Peng H.
        Hsa-let-7g promotes osteosarcoma by reducing HOXB1 to activate NF-kB pathway.
        Biomed. Pharmacother. 2019; 109: 2335-2341