Can Gollmann-Tepeköylü1,2, Leo Pölzl1,2, Michael Graber1,3, Jakob Hirsch1,2, Felix Nägele1,2, Daniela Lobenwein1,3, Michael W Hess4, Michael J Blumer3, Elke Kirchmair1,2, Johannes Zipperle2,5, Carina Hromada2,5, Severin Mühleder2,5, Hubert Hackl6, Martin Hermann7, Hemse Al Khamisi8, Martin Förster9, Michael Lichtenauer10, Rainer Mittermayr2,11, Patrick Paulus12, Helga Fritsch4, Nikolaos Bonaros1, Rudolf Kirchmair13, Joost P G Sluijter8, Sean Davidson14, Michael Grimm1, Johannes Holfeld1,2. 1. Department of Cardiac Surgery, Medical University of Innsbruck, Anichstrasse 35, 6020 Innsbruck, Austria. 2. Austrian Cluster for Tissue Regeneration, Vienna, Austria. 3. Division of Clinical and Functional Anatomy, Medical University of Innsbruck, Innsbruck, Austria. 4. Division of Histology and Embryology, Medical University of Innsbruck, Innsbruck, Austria. 5. Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Trauma Research Center, Vienna, Austria. 6. Division of Bioinformatics, Medical University of Innsbruck, Innsbruck, Austria. 7. Department of Anesthesiology, Medical University of Innsbruck, Innsbruck, Austria. 8. Laboratory of Experimental Cardiology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands. 9. Department of Cardiology, Pneumology and Angiology, Friedrich-Schiller-University Jena, Jena, Germany. 10. Department of Cardiology, Paracelsus Medical University Salzburg, Salzburg, Austria. 11. AUVA Trauma Center Meidling, Vienna, Austria. 12. Department of Anesthesiology and Operative Intensive Care Medicine, Kepler University Hospital Linz, Linz, Austria. 13. Department of Internal Medicine III, Medical University of Innsbruck, Innsbruck, Austria. 14. Hatter Cardiovascular Institute, University College London, London, UK.
Abstract
AIMS: As many current approaches for heart regeneration exert unfavourable side effects, the induction of endogenous repair mechanisms in ischaemic heart disease is of particular interest. Recently, exosomes carrying angiogenic miRNAs have been described to improve heart function. However, it remains challenging to stimulate specific release of reparative exosomes in ischaemic myocardium. In the present study, we sought to test the hypothesis that the physical stimulus of shock wave therapy (SWT) causes the release of exosomes. We aimed to substantiate the pro-angiogenic impact of the released factors, to identify the nature of their cargo, and to test their efficacy in vivo supporting regeneration and recovery after myocardial ischaemia. METHODS AND RESULTS: Mechanical stimulation of ischaemic muscle via SWT caused extracellular vesicle (EV) release from endothelial cells both in vitro and in vivo. Characterization of EVs via electron microscopy, nanoparticle tracking analysis and flow cytometry revealed specific exosome morphology and size with the presence of exosome markers CD9, CD81, and CD63. Exosomes exhibited angiogenic properties activating protein kinase b (Akt) and extracellular-signal regulated kinase (ERK) resulting in enhanced endothelial tube formation and proliferation. A miRNA array and transcriptome analysis via next-generation sequencing were performed to specify exosome content. miR-19a-3p was identified as responsible cargo, antimir-19a-3p antagonized angiogenic exosome effects. Exosomes and target miRNA were injected intramyocardially in mice after left anterior descending artery ligation. Exosomes resulted in improved vascularization, decreased myocardial fibrosis, and increased left ventricular ejection fraction as shown by transthoracic echocardiography. CONCLUSION: The mechanical stimulus of SWT causes release of angiogenic exosomes. miR-19a-3p is the vesicular cargo responsible for the observed effects. Released exosomes induce angiogenesis, decrease myocardial fibrosis, and improve left ventricular function after myocardial ischaemia. Exosome release via SWT could develop an innovative approach for the regeneration of ischaemic myocardium. Published on behalf of the European Society of Cardiology. All rights reserved.
AIMS: As many current approaches for heart regeneration exert unfavourable side effects, the induction of endogenous repair mechanisms in ischaemic heart disease is of particular interest. Recently, exosomes carrying angiogenic miRNAs have been described to improve heart function. However, it remains challenging to stimulate specific release of reparative exosomes in ischaemic myocardium. In the present study, we sought to test the hypothesis that the physical stimulus of shock wave therapy (SWT) causes the release of exosomes. We aimed to substantiate the pro-angiogenic impact of the released factors, to identify the nature of their cargo, and to test their efficacy in vivo supporting regeneration and recovery after myocardial ischaemia. METHODS AND RESULTS: Mechanical stimulation of ischaemic muscle via SWT caused extracellular vesicle (EV) release from endothelial cells both in vitro and in vivo. Characterization of EVs via electron microscopy, nanoparticle tracking analysis and flow cytometry revealed specific exosome morphology and size with the presence of exosome markers CD9, CD81, and CD63. Exosomes exhibited angiogenic properties activating protein kinase b (Akt) and extracellular-signal regulated kinase (ERK) resulting in enhanced endothelial tube formation and proliferation. A miRNA array and transcriptome analysis via next-generation sequencing were performed to specify exosome content. miR-19a-3p was identified as responsible cargo, antimir-19a-3p antagonized angiogenic exosome effects. Exosomes and target miRNA were injected intramyocardially in mice after left anterior descending artery ligation. Exosomes resulted in improved vascularization, decreased myocardial fibrosis, and increased left ventricular ejection fraction as shown by transthoracic echocardiography. CONCLUSION: The mechanical stimulus of SWT causes release of angiogenic exosomes. miR-19a-3p is the vesicular cargo responsible for the observed effects. Released exosomes induce angiogenesis, decrease myocardial fibrosis, and improve left ventricular function after myocardial ischaemia. Exosome release via SWT could develop an innovative approach for the regeneration of ischaemic myocardium. Published on behalf of the European Society of Cardiology. All rights reserved.
Authors: Can Gollmann-Tepeköylü; Felix Nägele; Michael Graber; Leo Pölzl; Daniela Lobenwein; Jakob Hirsch; Angela An; Regina Irschick; Bernhard Röhrs; Christian Kremser; Hubert Hackl; Rosalie Huber; Serena Venezia; David Hercher; Helga Fritsch; Nikolaos Bonaros; Nadia Stefanova; Ivan Tancevski; Dirk Meyer; Michael Grimm; Johannes Holfeld Journal: JCI Insight Date: 2020-08-06
Authors: Ricardo Cerqueira de Abreu; Hugo Fernandes; Paula A da Costa Martins; Susmita Sahoo; Costanza Emanueli; Lino Ferreira Journal: Nat Rev Cardiol Date: 2020-06-01 Impact factor: 32.419
Authors: Abraham Mendez-Fernandez; Hector A Cabrera-Fuentes; Bhaarathy Velmurugan; Jason Irei; William A Boisvert; Shengjie Lu; Derek J Hausenloy Journal: Cond Med Date: 2020-02