Peripheral blood mesenchymal stem cell-derived exosomes improve renal sympathetic denervation efficacy through β-catenin-mediated cardiac reprogramming

Abstract

Background: To investigate the role of self-peripheral blood mesenchymal stem cell (PBMSC)-derived exosomes (Exos) in enhancing renal sympathetic denervation (RD)-mediated heart regeneration following myocardial infarction (MI) in a porcine model.

Methods: Pigs (ejection fraction [EF] < 40% post-MI) were randomised to early sham RD or RD. At 2 weeks post-MI, autologous PBMSC-Exos were collected. At 30 days post-MI, pigs received either PBMSC-Exos (2 × 10¹³ particles) or phosphate-buffered saline and were followed until 90 days. Another cohort underwent myocardial biopsy at 14 days post-MI to assess PBMSC-Exos effects on ischaemic cardiomyocyte (CM) reprogramming, followed by adeno-associated viral therapy with miR-141-200-429 sponges or negative control sponges to explore the role of miR-141-200-429 clusters in reprogramming.

Results: Two weeks post-MI, RD hearts showed increased Exos uptake and inhibited the sympathetic nervous system. By 90 days, the RD+Exos group had 11%–26% higher EF than single-treatment groups (all p < .001), with improved survival and reduced fibrosis. Exos therapy enhanced RD effects by suppressing the renin‒angiotensin‒aldosterone system and transferring the miR-141-200-429 cluster into ischaemic CMs. CMs from RD-treated hearts cocultured with PBMSC-Exos^RD exhibited a more immature state, promoting reprogramming. β-Catenin overexpression further enhanced PBMSC-Exos^RD effects, while miR-141-200-429 inhibition blocked RD-induced CM reprogramming and survival. Ultimately, PBMSC-Exos^RD reduced dickkopf-1 (Dkk1) expression and activated GSK3β phosphorylation, thereby stimulating the Wnt/β-catenin pathway.

Conclusions: PBMSC-Exos^RD enhances RD-mediated cardiac repair through miR-141-200-429 cluster-dependent activation of the Wnt/β-catenin pathway, offering a novel therapeutic strategy for MI-induced heart failure. Our findings unveil a novel therapeutic strategy, highlighting that RD maintains its efficacy and safety when integrated with complementary approaches over extended periods.

Keywords

acute myocardial infarction / β-catenin / cardiac reprogramming / exosome / microRNA

References

[1] Azizi M, Sanghvi K, Saxena M, et al. Ultrasound renal denervation for hypertension resistant to a triple medication pill (RADIANCE-HTN TRIO): a randomised, multicentre, single-blind, sham-controlled trial. Lancet. 2021; 397: 2476-2486.

[2] Zhou Z, Liu C, Xu S, et al. Metabolism regulator adiponectin prevents cardiac remodeling and ventricular arrhythmias via sympathetic modulation in a myocardial infarction model. Basic Res Cardiol. 2022; 117: 34.

[3] Kwon JS, Schumacher SM, Gao E, et al. Characterization of βARKct engineered cellular extracellular vesicles and model specific cardioprotection. Am J Physiol Heart Circ Physiol. 2021; 320: H1276-H1289.

[4] Kimura K, Ieda M, Fukuda K. Development, maturation, and transdifferentiation of cardiac sympathetic nerves. Circ Res. 2012; 110: 325-336.

[5] Polhemus DJ, Trivedi RK, Sharp TE, et al. Repeated cell transplantation and adjunct renal denervation in ischemic heart failure: exploring modalities for improving cell therapy efficacy. Basic Res Cardiol. 2019; 114: 9.

[6] Böhm M, Ewen S, Wolf M. Renal denervation halts left ventricular remodeling and dysfunction in heart failure: new shores ahead. J Am Coll Cardiol. 2018; 72: 2622-2624.

[7] Sepe JJ, Gardner RT, Blake MR, et al. Therapeutics that promote sympathetic reinnervation modulate the inflammatory response after myocardial infarction. JACC Basic Transl Sci. 2022; 7: 915-930.

[8] Sharp TE, 3rdPolhemus DJ, Li Z, et al. Renal denervation prevents heart failure progression via inhibition of the renin‒angiotensin system. J Am Coll Cardiol. 2018; 72: 2609-26021.

[9] Garbern JC, Lee RT. Heart regeneration: 20 years of progress and renewed optimism. Dev Cell. 2022; 57: 424-439.

[10] Wang P, Deng Z, Li A, et al. β-Catenin promotes long-term survival and angiogenesis of peripheral blood mesenchymal stem cells via the Oct4 signaling pathway. Exp Mol Med. 2022; 54: 1434-1449.

[11] Sahoo S, Losordo DW. Exosomes and cardiac repair after myocardial infarction. Circ Res. 2014; 114: 333-344.

[12] Sharp TE, 3rdPolhemus DJ, Li Z, et al. Hypoxia induces heart regeneration in adult mice. Nature. 2017; 541: 222-227.

[13] Wang WE, Li L, Xia X, et al. Dedifferentiation, proliferation, and redifferentiation of adult mammalian cardiomyocytes after ischemic injury. Circulation. 2017; 136: 834-848.

[14] Li J, Sun S, Zhu D, et al. Inhalable stem cell exosomes promote heart repair after myocardial infarction. Circulation. 2024; 150(9): 710-723.

[15] Messerli FH, Bavishi C, Brguljan J, et al. Renal denervation in the antihypertensive arsenal—knowns and known unknowns. J Hypertens. 2022; 40(10): 1859-1875.

[16] Zhang S, Zhao L, Shen L, et al. Comparison of various niches for endothelial progenitor cell therapy on ischemic myocardial repair: coexistence of host collateralization and Akt-mediated angiogenesis produces a superior microenvironment. Arterioscler Thromb Vasc Biol. 2012; 32: 910-923.

[17] Peng Y, Zhao JL, Peng ZY, Xu WF, Yu GL. Exosomal miR-25-3p from mesenchymal stem cells alleviates myocardial infarction by targeting pro-apoptotic proteins and EZH2. Cell Death Dis. 2020; 11: 317.

[18] Théry C, Witwer KW, Aikawa E, et al. Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles. 2018; 7(1): 1535750.

[19] Hessvik NP, Sagini K, Romero S, et al. siRNA screening reveals that SNAP29 contributes to exosome release. Cell Mol Life Sci. 2023; 80(7): 177.

[20] Kumar R, Tang Q, Müller SA, et al. Fibroblast growth factor 2-mediated regulation of neuronal exosome release depends on VAMP3/cellubrevin in hippocampal neurons. Adv Sci. 2020; 7(6): 1902372.

[21] Tian Y, Liu Y, Wang T, et al. A microRNA-Hippo pathway that promotes cardiomyocyte proliferation and cardiac regeneration in mice. Sci Transl Med. 2015; 7: 279ra38.

[22] Patel V, Hajarnis S, Williams D, Hunter R, Huynh D, Igarashi P. MicroRNAs regulate renal tubule maturation through modulation of Pkd1. J Am Soc Nephrol. 2012; 23: 1941-1948.

[23] Pecot CV, Rupaimoole R, Yang D, et al. Tumour angiogenesis regulation by the miR-200 family. Nat Commun. 2013; 4: 2427.

[24] Akbar N, Razzaq SS, Salim A, Haneef K. Mesenchymal stem cell-derived exosomes and their microRNAs in heart repair and regeneration. J Cardiovasc Transl Res. 2024; 17(3): 505-522.

[25] Wan J, Lin S, Yu Z, et al. Protective effects of microRNA-200b-3p encapsulated by mesenchymal stem cells-secreted extracellular vesicles in myocardial infarction via regulating BCL2L11. J Am Heart Assoc. 2022; 11(12): e024330.

[26] Barrère-Lemaire S, Vincent A, Jorgensen C, Piot C, Nargeot J, Djouad F. Mesenchymal stromal cells for improvement of cardiac function following acute myocardial infarction: a matter of timing. Physiol Rev. 2024; 104(2): 659-725.

[27] Miller I, Min M, Yang C, et al. Ki67 is a graded rather than a binary marker of proliferation versus quiescence. Cell Rep. 2018; 24(5): 1105-1112. e5.

[28] Cho KW, Andrade M, Bae S, et al. Polycomb group protein CBX7 represses cardiomyocyte proliferation through modulation of the TARDBP/RBM38 axis. Circulation. 2023; 147(24): 1823-1842.

[29] Chen Y, Lüttmann FF, Schoger E, et al. Reversible reprogramming of cardiomyocytes to a fetal state drives heart regeneration in mice. Science. 2021; 373(6562): 1537-1540.

[30] Wang J, Pei B, Yan J, et al. hucMSC-derived exosomes alleviate the deterioration of colitis via the miR-146a/SUMO1 axis. Mol Pharm. 2022; 19(2): 484-493.

[31] Ibrahim AGE, Li C, Rogers R, et al. Augmenting canonical Wnt signalling in therapeutically inert cells converts them into therapeutically potent exosome factories. Nat Biomed Eng. 2019; 3(9): 695-705.

[32] Lian X, Zhang J, Azarin SM, et al. Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/β-catenin signaling under fully defined conditions. Nat Protoc. 2013; 8(1): 162-175.

[33] Zhu C, Yuan T, Krishnan J. Targeting cardiomyocyte cell cycle regulation in heart failure. Basic Res Cardiol. 2024; 119(3): 349-369.

[34] Gabisonia K, Prosdocimo G, Aquaro GD, et al. MicroRNA therapy stimulates uncontrolled cardiac repair after myocardial infarction in pigs. Nature. 2019; 569: 418-422.

[35] Dzau VJ, Hodgkinson CP. RNA therapeutics for the cardiovascular system. Circulation. 2024; 149(9): 707-716.

[36] Fiebeler A, Schmidt F, Müller DN, et al. Mineralocorticoid receptor affects AP-1 and nuclear factor-kappaB activation in angiotensin II-induced cardiac injury. Hypertension. 2001; 37(2 pt 2): 787-793.

[37] Li X, Wu F, Günther S, et al. Inhibition of fatty acid oxidation enables heart regeneration in adult mice. Nature. 2023; 622: 619-626.

[38] Liu J, Xiao Q, Xiao J, et al. Wnt/β-catenin signalling: function, biological mechanisms, and therapeutic opportunities. Signal Transduct Target Ther. 2022; 7: 3.

[39] Huang M, Wang X, Banerjee M, et al. Regulation of myogenesis by a Na/K-ATPase α1 caveolin-binding motif. Stem Cells. 2022; 40: 133-148.

[40] Deng F, Peng L, Li Z, et al. YAP triggers the Wnt/β-catenin signalling pathway and promotes enterocyte self-renewal, regeneration and tumorigenesis after DSS-induced injury. Cell Death Dis. 2018; 9: 153.

[41] Kim S, Song G, Lee T, et al. PARsylated transcription factor EB (TFEB) regulates the expression of a subset of Wnt target genes by forming a complex with β-catenin-TCF/LEF1. Cell Death Differ. 2021; 28: 2555-2570.

[42] Polhemus DJ, Gao J, Scarborough AL, et al. Radiofrequency renal denervation protects the ischemic heart via inhibition of GRK2 and increased nitric oxide signaling. Circ Res. 2016; 119: 470-480.

[43] Okamura K, Shimada H, Imazato K, et al. Impact of renal denervation on quality of life (How does renal denervation contribute to improving hypertension treatment affected by poor medication adherence?). Hypertens Res. 2024; 47: 2652-2658.

[44] Taru V, Szabo G, Mehal W, Reiberger T. Inflammasomes in chronic liver disease: hepatic injury, fibrosis progression and systemic inflammation. J Hepatol. 2024; 81(5): 895-910.

[45] Bhatt DL, Kandzari DE, O'Neill WW, et al. A controlled trial of renal denervation for resistant hypertension. N Engl J Med. 2014; 370: 1393-1401.

[46] Sharp TE, 3rdLefer DJ. Renal denervation to treat heart failure. Annu Rev Physiol. 2021; 83: 39-58.

[47] Davidson SM, Padró T, Bollini S, et al. Progress in cardiac research: from rebooting cardiac regeneration to a complete cell atlas of the heart. Cardiovasc Res. 2021; 117: 2161-2174.

[48] Feng Y, Wang Y, Li L, Yang Y, Tan X, Chen T. Exosomes induce crosstalk between multiple types of cells and cardiac fibroblasts: therapeutic potential for remodeling after myocardial infarction. Int J Nanomed. 2024; 19: 10605-10621.

[49] Mohamed TMA, Ang YS, Radzinsky E, et al. Regulation of cell cycle to stimulate adult cardiomyocyte proliferation and cardiac regeneration. Cell. 2018; 173: 104-116. e12.

[50] Gallet R, Dawkins J, Valle J, et al. Exosomes secreted by cardiosphere-derived cells reduce scarring, attenuate adverse remodelling, and improve function in acute and chronic porcine myocardial infarction. Eur Heart J. 2017; 38: 201-211.

[51] Gao L, Mei S, Zhang S, et al. Cardio-renal exosomes in myocardial infarction serum regulate proangiogenic paracrine signaling in adipose mesenchymal stem cells. Theranostics. 2020; 10: 1060-1073.

[52] Crewe C, Joffin N, Rutkowski JM, et al. An endothelial-to-adipocyte extracellular vesicle axis governed by metabolic state. Cell. 2018; 175(3): 695-708.

[53] Liang X, Li C, Song J, Liu A, et al. HucMSC-Exo promote mucosal healing in experimental colitis by accelerating intestinal stem cells and epithelium regeneration via Wnt signaling pathway. Int J Nanomed. 2023; 18: 2799-2818.

[54] Feng D, Yan K, Liang H, et al. CBP-mediated Wnt3a/β-catenin signaling promotes cervical oncogenesis initiated by Piwil2. Neoplasia. 2021; 23(1): 1-11.

[55] Kharaziha P, Ceder S, Li Q, Panaretakis T. Tumor cell-derived exosomes: a message in a bottle. Biochim Biophys Acta. 2012; 1826: 103-111.

[56] Wang J, Pei B, Yan J, et al. hucMSC-derived exosomes alleviate the deterioration of colitis via the miR-146a/SUMO1 axis. Mol Pharm. 2022; 19: 484-493.

[57] Zheng X, Li X, Lyu Y, He Y, Wan W, Jiang X. Renal sympathetic denervation in rats ameliorates cardiac dysfunction and fibrosis post-myocardial infarction involving microRNAs. Med Sci Monit. 2016; 22: 2751-2760.

[58] Gao L, Qiu F, Cao H, et al. Therapeutic delivery of microRNA-125a-5p oligonucleotides improves recovery from myocardial ischemia/reperfusion injury in mice and swine. Theranostics. 2023; 13: 685-703.

[59] Braga L, Ali H, Secco I, Giacca M. Non-coding RNA therapeutics for cardiac regeneration. Cardiovasc Res. 2021; 117: 674-693.

[60] Papukashvili D, Rcheulishvili N, Liu C, et al. Perspectives on miRNAs targeting DKK1 for developing hair regeneration therapy. Cells. 2021; 10: 2957.

[61] Li S, Yin Y, Yao L, et al. TNF-α treatment increases DKK1 protein levels in primary osteoblasts via upregulation of DKK1 mRNA levels and downregulation of miR-335-5p. Mol Med Rep. 2020; 22: 1017-1025.

[62] Liang XL, Wang YL, Wang PR. MiR-200a with CDC7 as a direct target declines cell viability and promotes cell apoptosis in Wilm's tumor via Wnt/β-catenin signaling pathway. Mol Cell Biochem. 2021; 476: 2409-2420.

[63] Kook SH, Heo JS, Lee JC. Crucial roles of canonical Runx2-dependent pathway on Wnt1-induced osteoblastic differentiation of human periodontal ligament fibroblasts. Mol Cell Biochem. 2015; 402: 213-223.

[64] Dharavath B, Butle A, Chaudhary A, et al. Recurrent UBE3C-LRP5 translocations in head and neck cancer with therapeutic implications. NPJ Precis Oncol. 2024; 8: 63.

[65] Frangogiannis NG. The inflammatory response in myocardial injury, repair, and remodelling. Nat Rev Cardiol. 2014; 11(5): 255-265.

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