Extracellular vesicles from human cardiac stromal cells up-regulate cardiomyocyte protective responses to hypoxia.
Apoptosis
Exosome
Mesenchymal stromal cell
Multi-omics
RNA-sequencing
miRNA
Journal
Stem cell research & therapy
ISSN: 1757-6512
Titre abrégé: Stem Cell Res Ther
Pays: England
ID NLM: 101527581
Informations de publication
Date de publication:
12 Oct 2024
12 Oct 2024
Historique:
received:
27
08
2024
accepted:
07
10
2024
medline:
13
10
2024
pubmed:
13
10
2024
entrez:
12
10
2024
Statut:
epublish
Résumé
Cell therapy can protect cardiomyocytes from hypoxia, primarily via paracrine secretions, including extracellular vesicles (EVs). Since EVs fulfil specific biological functions based on their cellular origin, we hypothesised that EVs from human cardiac stromal cells (CMSCLCs) obtained from coronary artery bypass surgery may have cardioprotective properties. This study characterises CMSCLC EVs (C_EVs), miRNA cargo, cardioprotective efficacy and transcriptomic modulation of hypoxic human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). C_EVs are compared to bone marrow mesenchymal stromal cell EVs (B_EVs) which are a known therapeutic EV type. Cells were characterised for surface markers, gene expression and differentiation potential. EVs were compared for yield, phenotype, and ability to protect hiPSC-CMs from hypoxia/reoxygenation injury. EV dose was normalised by both protein concentration and particle count, allowing direct comparison. C_EV and B_EV miRNA cargo was profiled and RNA-seq was performed on EV-treated hypoxic hiPSC-CMs, then data were integrated by multi-omics. Confirmatory experiments were carried out using miRNA mimics. At the same dose, C_EVs were more effective than B_EVs at protecting CM integrity, reducing apoptotic markers, and cell death during hypoxia. While C_EVs and B_EVs shared 70-77% similarity in miRNA content, C_EVs contained unique miRNAs, including miR-202-5p, miR-451a and miR-142-3p. Delivering miRNA mimics confirmed that miR-1260a and miR-202/451a/142 were cardioprotective, and the latter upregulated protective pathways similar to whole C_EVs. This study demonstrates the potential of cardiac tissues, routinely discarded following surgery, as a valuable source of EVs for myocardial infarction therapy. We also identify miR-1260a as protective of CM hypoxia.
Sections du résumé
BACKGROUND
BACKGROUND
Cell therapy can protect cardiomyocytes from hypoxia, primarily via paracrine secretions, including extracellular vesicles (EVs). Since EVs fulfil specific biological functions based on their cellular origin, we hypothesised that EVs from human cardiac stromal cells (CMSCLCs) obtained from coronary artery bypass surgery may have cardioprotective properties.
OBJECTIVES
OBJECTIVE
This study characterises CMSCLC EVs (C_EVs), miRNA cargo, cardioprotective efficacy and transcriptomic modulation of hypoxic human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). C_EVs are compared to bone marrow mesenchymal stromal cell EVs (B_EVs) which are a known therapeutic EV type.
METHODS
METHODS
Cells were characterised for surface markers, gene expression and differentiation potential. EVs were compared for yield, phenotype, and ability to protect hiPSC-CMs from hypoxia/reoxygenation injury. EV dose was normalised by both protein concentration and particle count, allowing direct comparison. C_EV and B_EV miRNA cargo was profiled and RNA-seq was performed on EV-treated hypoxic hiPSC-CMs, then data were integrated by multi-omics. Confirmatory experiments were carried out using miRNA mimics.
RESULTS
RESULTS
At the same dose, C_EVs were more effective than B_EVs at protecting CM integrity, reducing apoptotic markers, and cell death during hypoxia. While C_EVs and B_EVs shared 70-77% similarity in miRNA content, C_EVs contained unique miRNAs, including miR-202-5p, miR-451a and miR-142-3p. Delivering miRNA mimics confirmed that miR-1260a and miR-202/451a/142 were cardioprotective, and the latter upregulated protective pathways similar to whole C_EVs.
CONCLUSIONS
CONCLUSIONS
This study demonstrates the potential of cardiac tissues, routinely discarded following surgery, as a valuable source of EVs for myocardial infarction therapy. We also identify miR-1260a as protective of CM hypoxia.
Identifiants
pubmed: 39396003
doi: 10.1186/s13287-024-03983-y
pii: 10.1186/s13287-024-03983-y
doi:
Substances chimiques
MicroRNAs
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
363Subventions
Organisme : National Science and Technology Council (TW)
ID : 112-2636-B-038-005
Organisme : Office of Research and Development, Taipei Medical University
ID : 112-3805-006-111
Informations de copyright
© 2024. The Author(s).
Références
Barile L, Moccetti T, Marbán E, Vassalli G. Roles of exosomes in cardioprotection. Eur Heart J. 2017;38:1372–9.
pubmed: 27443883
Tariq U, Gupta M, Pathak S, Patil R, Dohare A, Misra SK. Role of Biomaterials in Cardiac Repair and Regeneration: therapeutic intervention for myocardial infarction. ACS Biomater Sci Eng. 2022;8:3271–98.
pubmed: 35867701
doi: 10.1021/acsbiomaterials.2c00454
Han MA, Jeon JH, Shin JY, Kim HJ, Lee JS, Seo CW, et al. Intramyocardial delivery of human cardiac stem cell spheroids with enhanced cell engraftment ability and cardiomyogenic potential for myocardial infarct repair. J Controlled Release. 2021;336:499–509.
doi: 10.1016/j.jconrel.2021.06.040
Oldershaw R, Owens WA, Sutherland R, Linney M, Liddle R, Magana L, et al. Human Cardiac-Mesenchymal Stem Cell-Like cells, a Novel Cell Population with therapeutic potential. Stem Cells Dev. 2019;28:593–607.
pubmed: 30803370
doi: 10.1089/scd.2018.0170
Nguyen H, Hsu C-C, Meeson A, Oldershaw R, Richardson G, Czosseck A, et al. Differentiation, metabolism, and cardioprotective secretory functions of human cardiac stromal cells from ischemic and Endocarditis patients. Stem Cells Dev. 2024. https://doi.org/10.1089/scd.2024.0103 .
doi: 10.1089/scd.2024.0103
pubmed: 38940748
Czosseck A, Chen MM, Nguyen H, Meeson A, Hsu C, Chen C, et al. Porous scaffold for mesenchymal cell encapsulation and exosome-based therapy of ischemic diseases. J Controlled Release. 2022;352:879–92.
doi: 10.1016/j.jconrel.2022.10.057
Kompa AR, Greening DW, Kong AM, McMillan PJ, Fang H, Saxena R, et al. Sustained subcutaneous delivery of secretome of human cardiac stem cells promotes cardiac repair following myocardial infarction. Cardiovasc Res. 2021;117:918–29.
pubmed: 32251516
doi: 10.1093/cvr/cvaa088
Chien KR, Frisén J, Fritsche-Danielson R, Melton DA, Murry CE, Weissman IL. Regenerating the field of cardiovascular cell therapy. Nat Biotechnol. 2019;37:232–7.
pubmed: 30778231
doi: 10.1038/s41587-019-0042-1
Davidson SM, Boulanger CM, Aikawa E, Badimon L, Barile L, Binder CJ, et al. Methods for the identification and characterization of extracellular vesicles in cardiovascular studies: from exosomes to microvesicles. Cardiovasc Res. 2022. https://doi.org/10.1093/cvr/cvac031 .
doi: 10.1093/cvr/cvac031
pubmed: 35916078
Adamiak M, Cheng G, Bobis-Wozowicz S, Zhao L, Kedracka-Krok S, Samanta A, et al. Induced Pluripotent Stem cell (iPSC)-derived extracellular vesicles are safer and more effective for cardiac repair than iPSCs. Circ Res. 2018;122:296–309.
pubmed: 29118058
doi: 10.1161/CIRCRESAHA.117.311769
Huang P, Wang L, Li Q, Xu J, Xu J, Xiong Y, et al. Combinatorial treatment of acute myocardial infarction using stem cells and their derived exosomes resulted in improved heart performance. Stem Cell Res Ther doi. 2019. https://doi.org/10.1186/s13287-019-1353-3 .
doi: 10.1186/s13287-019-1353-3
Gallet R, Dawkins J, Valle J, Simsolo E, De Couto G, Middleton R, 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;38:201–11.
pubmed: 28158410
Driedonks T, Jiang L, Carlson B, Han Z, Liu G, Queen SE, et al. Pharmacokinetics and biodistribution of extracellular vesicles administered intravenously and intranasally to Macaca nemestrina. J Extracell Biology. 2022;1:1–34.
doi: 10.1002/jex2.59
Livkisa D, Chang T, Burnouf T, Czosseck A, Le NTN, Shamrin G, et al. Extracellular vesicles purified from serum-converted human platelet lysates offer strong protection after cardiac ischaemia/reperfusion injury. Biomaterials. 2024;306:122502.
pubmed: 38354518
doi: 10.1016/j.biomaterials.2024.122502
Barile L, Lionetti V, Cervio E, Matteucci M, Gherghiceanu M, Popescu LM, et al. Extracellular vesicles from human cardiac progenitor cells inhibit cardiomyocyte apoptosis and improve cardiac function after myocardial infarction. Cardiovasc Res. 2014;103:530–41.
pubmed: 25016614
doi: 10.1093/cvr/cvu167
Ciullo A, Biemmi V, Milano G, Bolis S, Cervio E, Fertig ET, et al. Exosomal expression of CXCR4 targets cardioprotective vesicles to myocardial infarction and improves outcome after systemic administration. Int J Mol Sci Doi. 2019. https://doi.org/10.3390/ijms20030468 .
doi: 10.3390/ijms20030468
Vicencio JM, Yellon DM, Sivaraman V, Das D, Boi-Doku C, Arjun S, et al. Plasma exosomes protect the myocardium from ischemia-reperfusion injury. J Am Coll Cardiol. 2015;65:1525–36.
pubmed: 25881934
doi: 10.1016/j.jacc.2015.02.026
Luther KM, Haar L, McGuinness M, Wang Y, Lynch IVTL, Phan A, et al. Exosomal miR-21a-5p mediates cardioprotection by mesenchymal stem cells. J Mol Cell Cardiol. 2018;119:125–37.
pubmed: 29698635
doi: 10.1016/j.yjmcc.2018.04.012
Ikeda G, Santoso MR, Tada Y, Li AM, Vaskova E, Jung JH, et al. Mitochondria-Rich Extracellular vesicles from autologous stem cell–derived cardiomyocytes restore energetics of ischemic myocardium. J Am Coll Cardiol. 2021;77:1073–88.
pubmed: 33632482
doi: 10.1016/j.jacc.2020.12.060
van de Wakker SI, Bauzá-Martinez J, Ríos Arceo C, Manjikian H, Snijders Blok CJB, Roefs MT, et al. Size matters: functional differences of small extracellular vesicle subpopulations in cardiac repair responses. J Extracell Vesicles. 2024. https://doi.org/10.1002/jev2.12396 .
doi: 10.1002/jev2.12396
pubmed: 38868945
Lundy DJ, Szomolay B, Liao C-T. Systems approaches to Cell Culture-Derived Extracellular vesicles for acute kidney Injury Therapy: prospects and challenges. Function. 2024. https://doi.org/10.1093/function/zqae012 .
doi: 10.1093/function/zqae012
pubmed: 38706963
Shekari F, Alibhai FJ, Baharvand H, Börger V, Bruno S, Davies O, et al. Cell culture-derived extracellular vesicles: considerations for reporting cell culturing parameters. J Extracell Biology. 2023. https://doi.org/10.1002/jex2.115 .
doi: 10.1002/jex2.115
Ge X, Meng Q, Wei L, Liu J, Li M, Liang X, et al. Myocardial ischemia-reperfusion induced cardiac extracellular vesicles harbour proinflammatory features and aggravate heart injury. J Extracell Vesicles. 2021. https://doi.org/10.1002/jev2.12072 .
doi: 10.1002/jev2.12072
pubmed: 33664937
Biemmi V, Milano G, Ciullo A, Cervio E, Burrello J, Cas MD, et al. Inflammatory extracellular vesicles prompt heart dysfunction via TRL4-dependent NF-κB activation. Theranostics. 2020;10:2773–90.
pubmed: 32194834
doi: 10.7150/thno.39072
Song R, Dasgupta C, Mulder C, Zhang L. MicroRNA-210 controls mitochondrial metabolism and protects heart function in myocardial infarction. Circulation. 2022;145:1140–53.
pubmed: 35296158
doi: 10.1161/CIRCULATIONAHA.121.056929
Li J, Salvador AM, Li G, Valkov N, Ziegler O, Yeri A, et al. Mir-30d regulates Cardiac Remodeling by Intracellular and Paracrine Signaling. Circ Res. 2021;128:E1–23.
pubmed: 33092465
doi: 10.1161/CIRCRESAHA.120.317244
Xiao C, Wang K, Xu Y, Hu H, Zhang N, Wang Y, et al. Transplanted mesenchymal stem cells reduce autophagic flux in infarcted hearts via the exosomal transfer of miR-125b. Circ Res. 2018;123:564–78.
pubmed: 29921652
doi: 10.1161/CIRCRESAHA.118.312758
Yang L, Wang B, Zhou Q, Wang Y, Liu X, Liu Z, et al. MicroRNA-21 prevents excessive inflammation and cardiac dysfunction after myocardial infarction through targeting KBTBD7. Cell Death Dis doi. 2018. https://doi.org/10.1038/s41419-018-0805-5 .
doi: 10.1038/s41419-018-0805-5
Zhao J, Li X, Hu J, Chen F, Qiao S, Sun X, et al. Mesenchymal stromal cell-derived exosomes attenuate myocardial ischaemia-reperfusion injury through mir-182-regulated macrophage polarization. Cardiovasc Res. 2019;115:1205–16.
pubmed: 30753344
doi: 10.1093/cvr/cvz040
Pompilio G, Nigro P, Bassetti B, Capogrossi MC. Bone Marrow Cell Therapy for Ischemic Heart Disease. Circ Res. 2015;117:490–3.
pubmed: 26316604
doi: 10.1161/CIRCRESAHA.115.307184
Zhu LP, Tian T, Wang JY, He JN, Chen T, Pan M, et al. Hypoxia-elicited mesenchymal stem cell-derived exosomes facilitates cardiac repair through miR-125b-mediated prevention of cell death in myocardial infarction. Theranostics. 2018;8:6163–77.
pubmed: 30613290
doi: 10.7150/thno.28021
Yang H, Shao N, Holmström A, Zhao X, Chour T, Chen H, et al. Transcriptome analysis of non human primate-induced pluripotent stem cell-derived cardiomyocytes in 2D monolayer culture vs. 3D engineered heart tissue. Cardiovasc Res. 2021;117:2125–36.
pubmed: 33002105
doi: 10.1093/cvr/cvaa281
Barile L, Marbán E. Injury minimization after myocardial infarction: focus on extracellular vesicles. Eur Heart J. 2024;45:1602–9.
pubmed: 38366191
doi: 10.1093/eurheartj/ehae089
Zhao X, Chen H, Xiao D, Yang H, Itzhaki I, Qin X, et al. Comparison of non-human Primate versus Human Induced Pluripotent Stem cell-derived cardiomyocytes for treatment of myocardial infarction. Stem Cell Rep. 2018;10:422–35.
doi: 10.1016/j.stemcr.2018.01.002
Hidalgo A, Glass N, Ovchinnikov D, Yang SK, Zhang X, Mazzone S, et al. Modelling ischemia-reperfusion injury (IRI) in vitro using metabolically matured induced pluripotent stem cell-derived cardiomyocytes. APL Bioeng. 2018. https://doi.org/10.1063/1.5000746 .
doi: 10.1063/1.5000746
pubmed: 31069299
pmcid: 6481709
Mishra PK, Adameova A, Hill JA, Baines CP, Kang PM, Downey JM, et al. Guidelines for evaluating myocardial cell death. Am J Physiol Heart Circ Physiol. 2019;317:H891–922.
pubmed: 31418596
pmcid: 6879915
doi: 10.1152/ajpheart.00259.2019
Welsh JA, Goberdhan DCI, O’Driscoll L, Buzas EI, Blenkiron C, Bussolati B, et al. Minimal information for studies of extracellular vesicles (MISEV2023): from basic to advanced approaches. J Extracell Vesicles. 2024. https://doi.org/10.1002/jev2.12404 .
doi: 10.1002/jev2.12404
pubmed: 39140467
pmcid: 11322860
Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol doi. 2002. https://doi.org/10.1186/gb-2002-3-7-research0034 .
doi: 10.1186/gb-2002-3-7-research0034
Masè M, Grasso M, Avogaro L, D’Amato E, Tessarolo F, Graffigna A, et al. Selection of reference genes is critical for miRNA expression analysis in human cardiac tissue. A focus on atrial fibrillation. Sci Rep. 2017;7:1–10.
doi: 10.1038/srep41127
Viswanathan S, Shi Y, Galipeau J, Krampera M, Leblanc K, Martin I, et al. Mesenchymal stem versus stromal cells: International Society for Cell & Gene Therapy (ISCT
pubmed: 31526643
doi: 10.1016/j.jcyt.2019.08.002
Oldershaw RA, Richardson G, Carling P, Owens WA, Lundy DJ, Meeson A. Cardiac mesenchymal stem cell-like cells derived from a young patient with bicuspid aortic valve Disease have a prematurely aged phenotype. Biomedicines. 2022;10:3143.
pubmed: 36551899
doi: 10.3390/biomedicines10123143
Ward MC, Gilad Y. A generally conserved response to hypoxia in iPSC-derived cardiomyocytes from humans and chimpanzees. Elife. 2019;8:1–32.
doi: 10.7554/eLife.42374
Karbassi E, Fenix A, Marchiano S, Muraoka N, Nakamura K, Yang X, et al. Cardiomyocyte maturation: advances in knowledge and implications for regenerative medicine. Nat Rev Cardiol. 2020;17:341–59.
pubmed: 32015528
pmcid: 7239749
doi: 10.1038/s41569-019-0331-x
Lynch JM, Maillet M, Vanhoutte D, Schloemer A, Sargent MA, Blair NS, et al. A thrombospondin-dependent pathway for a protective ER stress response. Cell. 2012;149:1257–68.
pubmed: 22682248
doi: 10.1016/j.cell.2012.03.050
Akki A, Su J, Yano T, Gupta A, Wang Y, Leppo MK, et al. Creatine kinase overexpression improves ATP kinetics and contractile function in postischemic myocardium. Am J Physiol Heart Circ Physiol. 2012. https://doi.org/10.1152/ajpheart.00268.2012 .
doi: 10.1152/ajpheart.00268.2012
pubmed: 22886411
Yoshida T, Maulik N, Ho YS, Alam J, Das DK. Hmox-1 constitutes an adaptive response to effect antioxidant cardioprotection: a study with transgenic mice heterozygous for targeted disruption of the heme oxygenase-1 gene. Circulation. 2001;103:1695–701.
pubmed: 11273999
doi: 10.1161/01.CIR.103.12.1695
Lee DS, Chen J-H, Lundy DJ, Liu C-H, Hwang S-M, Pabon L, et al. Defined MicroRNAs induce aspects of maturation in mouse and human embryonic-stem-cell-derived cardiomyocytes. Cell Rep. 2015;12:1–8.
doi: 10.1016/j.celrep.2015.08.042
Wang X, Ha T, Zou J, Ren D, Liu L, Zhang X, et al. MicroRNA-125b protects against myocardial ischaemia/reperfusion injury via targeting p53-mediated apoptotic signalling and TRAF6. Cardiovasc Res. 2014;102:385–95.
pubmed: 24576954
doi: 10.1093/cvr/cvu044
Gao F, Kataoka M, Liu N, Liang T, Huang ZP, Gu F, et al. Therapeutic role of miR-19a/19b in cardiac regeneration and protection from myocardial infarction. Nat Commun Doi. 2019. https://doi.org/10.1038/s41467-019-09530-1 .
doi: 10.1038/s41467-019-09530-1
Lock MC, Tellam RL, Darby JRT, Soo JY, Brooks DA, Seed M, et al. Identification of novel miRNAs involved in Cardiac Repair following infarction in fetal and adolescent Sheep hearts. Front Physiol. 2020. https://doi.org/10.3389/fphys.2020.00614 .
doi: 10.3389/fphys.2020.00614
pubmed: 32587529
Li Y, Xu H, Fu X, Ji J, Shi Y, Wang Y. Upregulation of mir-202-5p promotes cell apoptosis and suppresses cell viability of hypoxia-induced myocardial H9c2 cells by targeting SOX6 to inhibit the activation of the PI3K/AKT/FOXO3a pathway. Int J Clin Exp Pathol. 2017;10:8884–94.
pubmed: 31966756
Li Y, Li Q, Zhang O, Guan X, Xue Y, Li S, et al. Mir-202‐5p protects rat against myocardial ischemia reperfusion injury by downregulating the expression of Trpv2 to attenuate the Ca2 + overload in cardiomyocytes. J Cell Biochem. 2019;120:13680–93.
pubmed: 31062423
doi: 10.1002/jcb.28641
Toldo S, Mauro AG, Narayan P, Kundur P, Neve F, La, Mezzaroma E, et al. Abstract 18896: plasma derived Alpha-2 macroglobulin limits the Inflammatory Injury in a mouse myocardial ischemia-reperfusion model. Circulation. 2017;136:A18896–18896.
Kitakaze M, Asakura M, Kim J, Shintani Y, Asanuma H, Hamasaki T, et al. Human atrial natriuretic peptide and nicorandil as adjuncts to reperfusion treatment for acute myocardial infarction (J-WIND): two randomised trials. Lancet. 2007;370:1483–93.
pubmed: 17964349
doi: 10.1016/S0140-6736(07)61634-1
Chin K, Kang G, Qu J, Gardner LB, Coetzee WA, Zito E, et al. The sarcoplasmic reticulum luminal thiol oxidase ERO1 regulates cardiomyocyte excitation-coupled calcium release and response to hemodynamic load. FASEB J. 2011;25:2583–91.
pubmed: 21507899
doi: 10.1096/fj.11-184622
Khachigian LM. Early growth response-1 in cardiovascular pathobiology. Circ Res. 2006;98:186–91.
pubmed: 16456111
doi: 10.1161/01.RES.0000200177.53882.c3
Zhu D, Zhang Z, Zhao J, Liu D, Gan L, Lau WB, et al. Targeting adiponectin receptor 1 phosphorylation against ischemic heart failure. Circ Res. 2022;131:E34–50.
pubmed: 35611695
doi: 10.1161/CIRCRESAHA.121.319976
Barile L, Cervio E, Lionetti V, Milano G, Ciullo A, Biemmi V, et al. Cardioprotection by cardiac progenitor cell-secreted exosomes: role of pregnancy-associated plasma protein-A. Cardiovasc Res. 2018;114:992–1005.
pubmed: 29518183
doi: 10.1093/cvr/cvy055
Nummi A, Nieminen T, Pätilä T, Lampinen M, Lehtinen ML, Kivistö S, et al. Epicardial delivery of autologous atrial appendage micrografts during coronary artery bypass surgery-safety and feasibility study. Pilot Feasibility Stud. 2017. https://doi.org/10.1186/s40814-017-0217-9 .
doi: 10.1186/s40814-017-0217-9
pubmed: 29276625