Adipose stem cell-derived exosomes circular RNA circFryl attenuate atrial fibrosis and cardiomyocyte apoptosis in atrial fibrillation

Articles in Press

Document Type : Original Article

Authors

Department of Cardiology, Xinghua People’s Hospital Affiliated to Yangzhou University, Xinghua, Jiangsu 225700, China

10.22038/ijbms.2025.84766.18341

Abstract

Objective(s): Atrial fibrillation (AF) is a prevalent arrhythmia accompanied by structural and electrical remodeling of the heart. Here, we examined the possible mechanisms behind the protective role of adipose-derived stem cell (ADSC)-derived exosomes in AF therapy.
Materials and Methods: We isolated exosomes from ADSCs. Exosome treatment was given. The left atrial diameter was measured by echocardiographic imaging. Cardiac fibrosis and damage were detected. The interaction between miR-338-3p with circFryl and tissue inhibitor of metalloproteinase mRNA (4TIMP4 mRNA) was predicted and investigated using qPCR and western blotting assay. 
Results: The overexpression of circFryl in ADSCs elevated the level of circFryl in exosomes and the myocytes, whereas knockdown of circFryl exhibited the opposite effects. Treatment with ADSC-exosomes significantly elevated circFryl level and recovered left atrial diameter, whereas knockdown of circFryl in exosomes abolished these effects. ADSC-exosomes alleviated the cardiac fibrosis and cell apoptosis in the AF model, and the knockdown of circFryl abolished these effects. ADSC-exosomes treatment suppressed viability and fibrosis and enhanced cell apoptosis in Ang-II-induced fibroblasts, which was reversed by depletion of circFryl. Online analysis of miRNA interaction targets showed potential binding between miR-338-3p with circFryl and TIMP4 mRNA. Knockdown of circFryl notably suppressed TIMP4 level, and inhibition of miR-338-3p recovered TIMP4 level. TIMP4 overexpression and miR-338-3p inhibition abolished the effects of sicircFryl in vitro and in vivo.  
Conclusion: The ADSC-derived exosomes delivered circFryl to interact with miR-338-3p, subsequently enhancing TIMP4 mRNA stability and expression in cardiac fibroblasts and myocytes. The circFryl/miR-338-3p/TIMP4 axis mediated the protective effects of ADSC on AF.

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References

1. Bizhanov KA, Аbzaliyev KB, Baimbetov AK, Sarsenbayeva AB, Lyan E. Atrial fibrillation: Epidemiology, pathophysiology, and clinical complications (literature review). J Cardiovasc Electrophysiol 2023; 34: 153-165.
2. Brundel BJJM, Ai X, Hills MT, Kuipers MF, Lip GYH, de Groot NMS. Atrial fibrillation. Nat Rev Dis Primers 2022; 8: 21.
3. Chyou JY, Barkoudah E, Dukes JW, Goldstein LB, Joglar JA, Lee AM, et al. Atrial fibrillation occurring during acute hospitalization: A scientific statement from the american heart association. Circulation 2023; 147: e676-e698.
4. Wijesurendra RS, Casadei B. Mechanisms of atrial fibrillation. Heart 2019; 105: 1860-1867.
5. Andersen JH, Andreasen L, Olesen MS. Atrial fibrillation-a complex polygenetic disease. Eur J Hum Genet 2021; 29: 1051-1060.
6. Kallistratos MS, Poulimenos LE, Manolis AJ. Atrial fibrillation and arterial hypertension. Pharmacol Res 2018; 128: 322-326.
7. Saleh K, Haldar S. Atrial fibrillation: A contemporary update. Clin Med (Lond) 2023; 23: 437-441.
8. Qin Y, Ge G, Yang P, Wang L, Qiao Y, Pan G, et al. An update on adipose-derived stem cells for regenerative medicine: Where challenge meets opportunity. Adv Sci (Weinh) 2023; 10: e2207334-2207361. 
9. Alió Del Barrio JL, De la Mata A, De Miguel MP, Arnalich-Montiel F, Nieto-Miguel T, El Zarif M, et al. Corneal regeneration using adipose-derived mesenchymal stem cells. Cells 2022; 11: 2549-2570.
10. Zhang M, Johnson-Stephenson TK, Wang W, Wang Y, Li J, Li L, et al. Mesenchymal stem cell-derived exosome-educated macrophages alleviate systemic lupus erythematosus by promoting efferocytosis and recruitment of IL-17+ regulatory T cell. Stem Cell Res Ther 2022; 13: 484-502.
11. Zhou C, Zhang B, Yang Y, Jiang Q, Li T, Gong J, et al. Stem cell-derived exosomes: Emerging therapeutic opportunities for wound healing. Stem Cell Res Ther 2023; 14: 107-125. 
12. Jin J, Shi Y, Gong J, Zhao L, Li Y, He Q, et al. Exosome secreted from adipose-derived stem cells attenuates diabetic nephropathy by promoting autophagy flux and inhibiting apoptosis in podocyte. Stem Cell Res Ther 2019; 10: 95-110.
13. Bacakova L, Zarubova J, Travnickova M, Musilkova J, Pajorova J, Slepicka P, et al. Stem cells: Their source, potency and use in regenerative therapies with focus on adipose-derived stem cells - a review. Biotechnol Adv 2018; 36: 1111-1126.
14. An Y, Lin S, Tan X, Zhu S, Nie F, Zhen Y, et al. Exosomes from adipose-derived stem cells and application to skin wound healing. Cell Prolif 2021; 54: e12993-13008.
15. Al-Ghadban S, Bunnell BA. Adipose tissue-derived stem cells: Immunomodulatory effects and therapeutic potential. Physiology (Bethesda) 2020; 35: 125-133.
16. Shen K, Wang X, Wang Y, Jia Y, Zhang Y, Wang K, et al. miR-125b-5p in adipose derived stem cells exosome alleviates pulmonary microvascular endothelial cells ferroptosis via Keap1/Nrf2/GPX4 in sepsis lung injury. Redox Biol 2023; 62: 102655-102672.
17. Li Q, Hu W, Huang Q, Yang J, Li B, Ma K, et al. MiR146a-loaded engineered exosomes released from silk fibroin patch promote diabetic wound healing by targeting IRAK1. Signal Transduct Target Ther 2023; 8: 62-75.
18. Yan C, Chen J, Wang C, Yuan M, Kang Y, Wu Z, et al. Milk exosomes-mediated miR-31-5p delivery accelerates diabetic wound healing through promoting angiogenesis. Drug Deliv 2022; 29: 214-228.
19. Youn SW, Li Y, Kim YM, Sudhahar V, Abdelsaid K, Kim HW, et al. Modification of cardiac progenitor cell-derived exosomes by miR-322 provides protection against myocardial infarction through Nox2-dependent angiogenesis. Antioxidants (Basel) 2019; 8: 18-33.
20. Greening DW, Xu R, Ji H, Tauro BJ, Simpson RJ. A protocol for exosome isolation and characterization: Evaluation of ultracentrifugation, density-gradient separation, and immunoaffinity capture methods. Methods Mol Biol 2015; 1295: 179-209.
21. Schüttler D, Bapat A, Kääb S, Lee K, Tomsits P, Clauss S, et al. Animal models of atrial fibrillation. Circ Res 2020; 127: 91-110.
22. Hahn RT, Leipsic J, Douglas PS, Jaber WA, Weissman NJ, Pibarot P, et al. Comprehensive echocardiographic assessment of normal transcatheter valve function. JACC Cardiovasc Imaging 2019; 12: 25-34.
23. Feng J, Li Y, Nie Y. Methods of mouse cardiomyocyte isolation from postnatal heart. J Mol Cell Cardiol 2022; 168: 35-43.
24. da Silva AMG, Cruz MS, de Souza KSC, Silbiger VN. Long non-coding RNA and circular RNA: new perspectives for molecular pathophysiology of atrial fibrillation. Mol Biol Rep 2023; 50: 2835-2845.
25. Zhu X, Tang X, Chong H, Cao H, Fan F, Pan J, et al. Expression profiles of circular RNA in human atrial fibrillation with valvular heart diseases. Front Cardiovasc Med 2020; 7: 597932-597942.
26. Wu N, Li J, Chen X, Xiang Y, Wu L, Li C, et al. Identification of long non-coding RNA and circular RNA expression profiles in atrial fibrillation. Heart Lung Circ 2020; 29: e157-e167.
27. Hao H, Yan S, Zhao X, Han X, Fang N, Zhang Y, et al. Atrial myocyte-derived exosomal microRNA contributes to atrial fibrosis in atrial fibrillation. J Transl Med 2022; 20: 407.
28. Wang Y, Liu B. Circular RNA in diseased heart. Cells 2020; 9: 1240-1257.
29. Hu X, Chen L, Wu S, Xu K, Jiang W, Qin M, et al. Integrative analysis reveals key circular RNA in atrial fibrillation. Front Genet 2019; 10: 108-120.
30. Hu X, Qin H, Yan Y, Wu W, Gong S, Wang L, et al. Exosomal circular RNAs: Biogenesis, effect, and application in cardiovascular diseases. Front Cell Dev Biol 2022; 10: 948256-948268.
31. Liu X, Wu M, He Y, Gui C, Wen W, Jiang Z, et al. Construction and integrated analysis of the ceRNA network hsa_circ_0000672/miR-516a-5p/TRAF6 and its potential function in atrial fibrillation. Sci Rep 2023; 13: 7701-7716.
32. Wu N, Li C, Xu B, Xiang Y, Jia X, Yuan Z, et al. Circular RNA mmu_circ_0005019 inhibits fibrosis of cardiac fibroblasts and reverses electrical remodeling of cardiomyocytes. BMC Cardiovasc Disord 2021; 21: 308-320.
33. Shen W, Zhao X, Li S. Exosomes derived from ADSCs attenuate sepsis-induced lung injury by delivery of Circ-Fryl and regulation of the miR-490-3p/SIRT3 pathway. Inflammation 2022; 45: 331-342.
34. Moore L, Fan D, Basu R, Kandalam V, Kassiri Z. Tissue inhibitor of metalloproteinases (TIMPs) in heart failure. Heart Fail Rev 2012; 17: 693-706. 
35. Xiao H, Lei H, Qin S, Ma K, Wang X. TGF-beta1 expression and atrial myocardium fibrosis increase in atrial fibrillation secondary to rheumatic heart disease. Clin Cardiol 2010; 33: 149-156.
36. Kim SK, Park JH, Kim JY, Choi JI, Joung B, Lee MH, et al. High plasma concentrations of transforming growth factor-β and tissue inhibitor of metalloproteinase-1: Potential non-invasive predictors for electroanatomical remodeling of atrium in patients with non-valvular atrial fibrillation. Circ J 2011; 75: 557-564.
37. Gramley F, Lorenzen J, Koellensperger E, Kettering K, Weiss C, Munzel T. Atrial fibrosis and atrial fibrillation: The role of the TGF-β1 signaling pathway. Int J Cardiol 2010; 143: 405-413.
38. Rahmutula D, Marcus GM, Wilson EE, Ding CH, Xiao Y, Paquet AC, et al. Molecular basis of selective atrial fibrosis due to overexpression of transforming growth factor-β1. Cardiovasc Res 2013; 99: 769-779.
39. Verheule S, Sato T, Everett T 4th, Engle SK, Otten D, Rubart-von der Lohe M, et al. Increased vulnerability to atrial fibrillation in transgenic mice with selective atrial fibrosis caused by overexpression of TGF-beta1. Circ Res 2004; 94: 1458-1465.
Iranian Journal of Basic Medical Sciences

Articles in Press, Accepted Manuscript
Available Online from 22 April 2025

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