Rabea Hinkel1, Deepak Ramanujam2, Veronika Kaczmarek3, Andrea Howe3, Katharina Klett3, Christina Beck2, Anne Dueck2, Thomas Thum4, Karl-Ludwig Laugwitz5, Lars Maegdefessel6, Christian Weber7, Christian Kupatt8, Stefan Engelhardt9. 1. 1. Medizinische Klinik und Poliklinik, Klinikum Rechts der Isar, Technische Universität München, Munich, Germany; Institute for Cardiovascular Prevention, Ludwigs-Maximilians-Universität München, Munich, Germany; Deutsches Primatenzentrum GmbH, Leibnitz-Institut für Primatenforschung, Laboratory Animal Science Unit, Göttingen, Germany; DZHK (German Center for Cardiovascular Research), partner site Munich Heart Alliance, Munich, Germany. 2. DZHK (German Center for Cardiovascular Research), partner site Munich Heart Alliance, Munich, Germany; Institut für Pharmakologie und Toxikologie, Technische Universität München, Munich, Germany. 3. 1. Medizinische Klinik und Poliklinik, Klinikum Rechts der Isar, Technische Universität München, Munich, Germany; Institute for Cardiovascular Prevention, Ludwigs-Maximilians-Universität München, Munich, Germany. 4. Institute of Molecular and Translational Therapeutic Strategies, Hannover Medical School, Hannover, Germany. 5. 1. Medizinische Klinik und Poliklinik, Klinikum Rechts der Isar, Technische Universität München, Munich, Germany; DZHK (German Center for Cardiovascular Research), partner site Munich Heart Alliance, Munich, Germany. 6. DZHK (German Center for Cardiovascular Research), partner site Munich Heart Alliance, Munich, Germany; Department of Vascular Surgery, Klinikum Rechts der Isar, Technische Universität München, Munich, Germany. 7. Institute for Cardiovascular Prevention, Ludwigs-Maximilians-Universität München, Munich, Germany; DZHK (German Center for Cardiovascular Research), partner site Munich Heart Alliance, Munich, Germany; Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University, Maastricht, the Netherlands. 8. 1. Medizinische Klinik und Poliklinik, Klinikum Rechts der Isar, Technische Universität München, Munich, Germany; DZHK (German Center for Cardiovascular Research), partner site Munich Heart Alliance, Munich, Germany. Electronic address: christian.kupatt@tum.de. 9. DZHK (German Center for Cardiovascular Research), partner site Munich Heart Alliance, Munich, Germany; Institut für Pharmakologie und Toxikologie, Technische Universität München, Munich, Germany. Electronic address: stefan.engelhardt@tum.de.
Abstract
BACKGROUND: miR-21 is a central regulator of cardiac fibrosis, and its inhibition in small-animal models has been shown to be an effective antifibrotic strategy in various organs, including the heart. Effective delivery of therapeutic antisense micro-ribonucleic acid (antimiR) molecules to the myocardium in larger organisms is challenging, though, and remains to be established for models of chronic heart failure. OBJECTIVES: The aims of this study were to test the applicability and therapeutic efficacy of local, catheter-based delivery of antimiR-21 in a pig model of heart failure and determine its effect on the cardiac transcriptomic signature and cellular composition. METHODS: Pigs underwent transient percutaneous occlusion of the left coronary artery and were followed up for 33 days. AntimiR-21 (10 mg) was applied by intracoronary infusion at days 5 and 19 after the injury. Cardiac function was assessed in vivo, followed by histological analyses and deep ribonucleic acid sequencing (RNA-seq) of the myocardium and genetic deconvolution analysis. RESULTS: AntimiR-21 effectively suppressed the remodeling-associated increase of miR-21. At 33 days after ischemia/reperfusion injury, LNA-21-treated hearts exhibited reduced cardiac fibrosis and hypertrophy and improved cardiac function. Deep RNA-seq revealed a significant derepression of the miR-21 targetome in antimiR-21-treated myocardium and a suppression of the inflammatory response and mitogen-activated protein kinase signaling. A genetic deconvolution approach built on deep RNA-seq and single-cell RNA-seq data identified reductions in macrophage and fibroblast numbers as the key cell types affected by antimiR-21 treatment. CONCLUSIONS: This study provides the first evidence for the feasibility and therapeutic efficacy of miR-21 inhibition in a large animal model of heart failure.
BACKGROUND:miR-21 is a central regulator of cardiac fibrosis, and its inhibition in small-animal models has been shown to be an effective antifibrotic strategy in various organs, including the heart. Effective delivery of therapeutic antisense micro-ribonucleic acid (antimiR) molecules to the myocardium in larger organisms is challenging, though, and remains to be established for models of chronic heart failure. OBJECTIVES: The aims of this study were to test the applicability and therapeutic efficacy of local, catheter-based delivery of antimiR-21 in a pig model of heart failure and determine its effect on the cardiac transcriptomic signature and cellular composition. METHODS:Pigs underwent transient percutaneous occlusion of the left coronary artery and were followed up for 33 days. AntimiR-21 (10 mg) was applied by intracoronary infusion at days 5 and 19 after the injury. Cardiac function was assessed in vivo, followed by histological analyses and deep ribonucleic acid sequencing (RNA-seq) of the myocardium and genetic deconvolution analysis. RESULTS: AntimiR-21 effectively suppressed the remodeling-associated increase of miR-21. At 33 days after ischemia/reperfusion injury, LNA-21-treated hearts exhibited reduced cardiac fibrosis and hypertrophy and improved cardiac function. Deep RNA-seq revealed a significant derepression of the miR-21 targetome in antimiR-21-treated myocardium and a suppression of the inflammatory response and mitogen-activated protein kinase signaling. A genetic deconvolution approach built on deep RNA-seq and single-cell RNA-seq data identified reductions in macrophage and fibroblast numbers as the key cell types affected by antimiR-21 treatment. CONCLUSIONS: This study provides the first evidence for the feasibility and therapeutic efficacy of miR-21 inhibition in a large animal model of heart failure.
Authors: Mikko Hänninen; Toni Jäntti; Heli Tolppanen; Heli Segersvärd; Tuukka Tarvasmäki; Johan Lassus; Mélanie Vausort; Yvan Devaux; Alessandro Sionis; Ilkka Tikkanen; Veli-Pekka Harjola; Päivi Lakkisto Journal: Int J Mol Sci Date: 2020-10-26 Impact factor: 5.923