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Original article
Published online: 2024-08-12
Enhanced therapeutic efficacy of SERCA2a gene-modified adipose-derived mesenchymal stem cell exosomes in doxorubicin-induced cardiomyopathy in male albino rats
DOI: 10.5603/fm.101080
Pubmed: 39136373
Abstract
Background: Worldwide, cancer is still the primary cause of death, and one of the most widely used anthracyclines for treating cancer is doxorubicin (DOX). But a major worry is DOX-induced cardiomyopathy, which is primarily resulted from an excess of reactive oxygen species. Heart sarcoplasmic reticulum calcium ion ATPase2a (SERCA2a) controls the amount of calcium ions stored in the sarcoplasmic reticulum (SR). This study aims to evaluate and compare the efficacy of SERCA2a gene modified adipose mesenchymal stem cell-derived exosomes (AMSCs-dE) to nontransfected AMSCs-dE, in DOX induced cardiomyopathy in adult male albino rat.
Materials and methods: Thirty one adult male albino rats were randomly divided into control group and DOX group that further subdivided into three DOX, AMSCs-dE and SERCA2a AMSCs-dE subgroups. AMSCs-dE were administered intravenously (IV). The levels of serum creatine kinase MB (CK-MB) were assessed after DOX injection and before sacrifice. Cardiac muscle samples were taken for histological analysis using Masson’s trichrome and hematoxylin and eosin stains two months after the experiment. Proliferating cell nuclear antigen (PCNA) and connexin 43 were stained using immunohistochemistry. The expression of TNF and SERCA2a genes and proteins was measured by real-time polymerase chain reaction (PCR) and Western blot (Wb) analysis, respectively. Fluorescent microscopy demonstrated non-transfected and transfected exosomes labeled with PKH26 and GFP, respectively, in culture and cardiac muscle.
Results: DOX induced myocarditis progressing to degenerative and fibrotic changes in cardiac muscle that regressed in response to AMSCs-dE therapy. However, SERCA2a gene modified AMSCs-dE treatment reversed the mentioned parameters to nearly its normal level.
Conclusions: These findings suggest that SERCA2a gene modification enhances the therapeutic efficacy of AMSCs-dE in treating DOX-induced cardiomyopathy.
Keywords: DOXAMSCsSERCA2aPCNAexosomesTNF
References
- Abouelnaga H, El-Khateeb D, Moemen Y, et al. Characterization of mesenchymal stem cells isolated from Wharton’s jelly of the human umbilical cord. Egypt Liver J. 2022; 12(1): 28–36.
- Angsutararux P, Luanpitpong S, Issaragrisil S. Chemotherapy-induced cardiotoxicity: overview of the roles of oxidative stress. Oxid Med Cell Longev. 2015; 2015: 795602.
- Samy W, Yousef M, El-Azab N, et al. Possible therapeutic effect of ghrelin and exosomes derived from mesenchymal stem cells on induced chronic pancreatitis of adult albino rats (histological and immunohistochemical study). Benha Medical Journal. 2022; 0(0): 0–0.
- Bancroft JD, Gamble M. Connective tissue stains. In: Bancroft JD, Gamble M. ed. Theory and Practice of Histological Techniques (6th ed). Churchill Livingstone, Edinburgh 2007.
- Bugrova AE, Strelnikova PA, Indeykina MI, et al. The dynamics of β-amyloid proteoforms accumulation in the brain of a 5xFAD mouse model of Alzheimer's disease. Int J Mol Sci. 2021; 23(1).
- de Carvalho TG, Pellenz FM, Laureano A, et al. A simple protocol for transfecting human mesenchymal stem cells. Biotechnol Lett. 2018; 40(3): 617–622.
- Christidi E, Brunham LR. Regulated cell death pathways in doxorubicin-induced cardiotoxicity. Cell Death Dis. 2021; 12(4): 339.
- Ebrahim N, El-Halim HE, Helal OK, et al. Effect of bone marrow mesenchymal stem cells-derived exosomes on diabetes-induced retinal injury: Implication of Wnt/ b-catenin signaling pathway. Biomed Pharmacother. 2022; 154: 113554.
- El-Derany MO, AbdelHamid SG. Bone marrow mesenchymal stem cells and their derived extracellular vesicles attenuate non-alcoholic steatohepatitis-induced cardiotoxicity via modulating cardiac mechanisms. Life (Basel). 2022; 12(3): 355.
- El-Sayed ESM, Mansour AM, Abdul-Hameed MS. Thymol and Carvacrol Prevent Doxorubicin-Induced Cardiotoxicity by Abrogation of Oxidative Stress, Inflammation, and Apoptosis in Rats. J Biochem Mol Toxicol. 2016; 30(1): 37–44.
- Femminò S, Bonelli F, Brizzi MF. Extracellular vesicles in cardiac repair and regeneration: Beyond stem-cell-based approaches. Front Cell Dev Biol. 2022; 10: 996887.
- García-Pedraza JÁ, García-Domingo M, Gómez-Roso M, et al. Hypertension exhibits 5-HT receptor as a modulator of sympathetic neurotransmission in the rat mesenteric vasculature. Hypertens Res. 2019; 42(5): 618–627.
- González-Cubero E, González-Fernández ML, Gutiérrez-Velasco L, et al. Isolation and characterization of exosomes from adipose tissue-derived mesenchymal stem cells. J Anat. 2021; 238(5): 1203–1217.
- Guo Q, Guo J, Yang R, et al. Cyclovirobuxine D attenuates doxorubicin-induced cardiomyopathy by suppression of oxidative damage and mitochondrial biogenesis impairment. Oxid Med Cell Longev. 2015; 2015: 151972.
- Kiernan JA. Histological and histochemical methods: theory and practice (3rd ed). Arnold Publisher, London 2001: 111–162.
- Kruse JM, Enghard P, Schröder T, et al. Weak diagnostic performance of troponin, creatine kinase and creatine kinase-MB to diagnose or exclude myocardial infarction after successful resuscitation. Int J Cardiol. 2014; 173(2): 216–221.
- Liu G, Liu Y, Wang R, et al. Spironolactone attenuates doxorubicin-induced cardiotoxicity in rats. Cardiovasc Ther. 2016; 34(4): 216–224.
- Liu S, Lan Y, Zhao Y, et al. Expression of connexin 43 protein in cardiomyocytes of heart failure mouse model. Front Cardiovasc Med. 2022; 9: 1028558.
- Liu W, Xu S, Liang S, et al. Hypertensive vascular and cardiac remodeling protection by allicin in spontaneous hypertension rats via CaMK Ⅱ/NF-κB pathway. Biomed Pharmacother. 2022; 155: 113802.
- Lodewijks F, McKinsey TA, Robinson EL. Fat-to-heart crosstalk in health and disease. Front Genet. 2023; 14: 990155.
- Lokman Hakim NY, Noble S, Thomas NV, et al. Genetic modification as a new approach to ameliorate the therapeutic efficacy of stem cells in diabetic retinopathy. Eur J Ophthalmol. 2022 [Epub ahead of print]: 11206721211073430.
- Lucero CM, Andrade DC, Toledo C, et al. Cardiac remodeling and arrhythmogenesis are ameliorated by administration of Cx43 mimetic peptide Gap27 in heart failure rats. Sci Rep. 2020; 10(1): 6878.
- Ma S, Li X, Dong L, et al. Protective effect of Sheng-Mai Yin, a traditional Chinese preparation, against doxorubicin-induced cardiac toxicity in rats. BMC Complement Altern Med. 2016; 16: 61.
- Mandziuk S, Gieroba R, Korga A, et al. The differential effects of green tea on dose-dependent doxorubicin toxicity. Food Nutr Res. 2015; 59: 29754.
- Mitry MA, Edwards JG. Doxorubicin induced heart failure: Phenotype and molecular mechanisms. Int J Cardiol Heart Vasc. 2016; 10: 17–24.
- Mullard A. Addressing cancer's grand challenges. Nat Rev Drug Discov. 2020; 19(12): 825–826.
- Sabry D, Marzouk S, Zakaria R, et al. The effect of exosomes derived from mesenchymal stem cells in the treatment of induced type 1 diabetes mellitus in rats. Biotechnol Lett. 2020; 42(8): 1597–1610.
- Saeed NM, El-Naga RN, El-Bakly WM, et al. Epigallocatechin-3-gallate pretreatment attenuates doxorubicin-induced cardiotoxicity in rats: a mechanistic study. Biochem Pharmacol. 2015; 95(3): 145–155.
- Sahu BD, Mahesh Kumar J, Sistla R. Baicalein, a bioflavonoid, prevents cisplatin-induced acute kidney injury by up-regulating antioxidant defenses and down-regulating the MAPKs and NF-κB pathways. PLoS One. 2015; 10(7): e0134139.
- Sariyer I. Transfection of neuronal cultures. Methods Mol Biol. 2021; 2311: 147–153.
- Percie du Sert N, Ahluwalia A, Alam S, et al. Reporting animal research: explanation and elaboration for the ARRIVE guidelines 2.0. PLoS Biol. 2020; 18(7): e3000411.
- Sheahan BJ, Freeman AN, Keeley TM, et al. Epithelial regeneration after doxorubicin arises primarily from early progeny of active intestinal stem cells. Cell Mol Gastroenterol Hepatol. 2021; 12(1): 119–140.
- Sheta A, Elsakkar M, Hamza M, et al. Effect of metformin and sitagliptin on doxorubicin-induced cardiotoxicity in adult male albino rats. Hum Exp Toxicol. 2016; 35(11): 1227–1239.
- Shi H, Qi C, Meng L, et al. Bone marrow-derived mesenchymal stem cells promote Helicobacter pylori-associated gastric cancer progression by secreting thrombospondin-2. Cell Prolif. 2021; 54(10): e13114.
- Sikkel MB, Hayward C, MacLeod KT, et al. SERCA2a gene therapy in heart failure: an antiarrythmic positive inotrope. Br J Pharmacol. 2014; 171(1): 38–54.
- Sun SJ, Wei R, Li F, et al. Mesenchymal stromal cell-derived exosomes in cardiac regeneration and repair. Stem Cell Reports. 2021; 16(7): 1662–1673.
- Takehara Y, Yabuuchi A, Ezoe K, et al. The restorative effects of adipose-derived mesenchymal stem cells on damaged ovarian function. Lab Invest. 2013; 93(2): 181–193.
- Torre E, Arici M, Lodrini AM, et al. SERCA2a stimulation by istaroxime improves intracellular Ca2+ handling and diastolic dysfunction in a model of diabetic cardiomyopathy. Cardiovasc Res. 2022; 118(4): 1020–1032.
- Wang K, Xu BC, Duan HY, et al. Late cardioprotection of exercise preconditioning against exhaustive exercise-induced myocardial injury by up-regulatation of connexin 43 expression in rat hearts. Asian Pac J Trop Med. 2015; 8(8): 658–663.
- Wang S, Zhou Y, Luo Y, et al. SERCA2a ameliorates cardiomyocyte T-tubule remodeling via the calpain/JPH2 pathway to improve cardiac function in myocardial ischemia/reperfusion mice. Sci Rep. 2021; 11(1): 2037.
- Wei W, Guo X, Gu L, et al. Bone marrow mesenchymal stem cell exosomes suppress phosphate-induced aortic calcification via SIRT6-HMGB1 deacetylation. Stem Cell Res Ther. 2021; 12(1): 235.
- Yang L, Deng J, Ma W, et al. Ablation of lncRNA attenuates pathological hypertrophy and heart failure. Theranostics. 2021; 11(16): 7995–8007.
- Yaşar M, Çakmak H, Dündar S, et al. The role of microRNAs in corneal neovascularization and its relation to VEGF. Cutan Ocul Toxicol. 2020; 39(4): 341–347.
- Zhao H, Li Z, Wang Y, et al. Bioengineered MSC-derived exosomes in skin wound repair and regeneration. Front Cell Dev Biol. 2023; 11: 1029671.
- Zhihao L, Jingyu Ni, Lan Li, et al. SERCA2a: a key protein in the Ca cycle of the heart failure. Heart Fail Rev. 2020; 25(3): 523–535.
- Zhou Y, Wen LL, Li YF, et al. Exosomes derived from bone marrow mesenchymal stem cells protect the injured spinal cord by inhibiting pericyte pyroptosis. Neural Regen Res. 2022; 17(1): 194–202.
- Zickri MB, Zaghloul S, Farouk M, et al. Effect of stem cell therapy on adriamycin induced tubulointerstitial injury. Int J Stem Cells. 2012; 5(2): 130–139.
- Zuris JA, Thompson DB, Shu Y, et al. Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol. 2015; 33(1): 73–80.
