AntimiR-21 Prevents Myocardial Dysfunction in a Pig Model of Ischemia/Reperfusion Injury

Rabea Hinkel; Deepak Ramanujam; Veronika Kaczmarek; Andrea Howe; Katharina Klett; Christina Beck; Anne Dueck; Thomas Thum; Karl-Ludwig Laugwitz; Lars Maegdefessel; Christian Weber; Christian Kupatt; Stefan Engelhardt

doi:10.1016/j.jacc.2020.02.041

AntimiR-21 Prevents Myocardial Dysfunction in a Pig Model of Ischemia/Reperfusion Injury

Rabea Hinkel, Deepak Ramanujam, Veronika Kaczmarek, Andrea Howe, Katharina Klett, Christina Beck, Anne Dueck, Thomas Thum, Karl-Ludwig Laugwitz, Lars Maegdefessel, Christian Weber, Christian Kupatt^*, Stefan Engelhardt^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

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. (C) 2020 The Authors. Published by Elsevier on behalf of the American College of Cardiology Foundation.

Original language	English
Pages (from-to)	1788-1800
Number of pages	13
Journal	Journal of the American College of Cardiology
Volume	75
Issue number	15
DOIs	https://doi.org/10.1016/j.jacc.2020.02.041
Publication status	Published - 21 Apr 2020

Keywords

cardiac disease
fibrosis
microRNA
miR-21
porcine model of heart failure
MICRORNA-21

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Hinkel, R., Ramanujam, D., Kaczmarek, V., Howe, A., Klett, K., Beck, C., Dueck, A., Thum, T., Laugwitz, K-L., Maegdefessel, L., Weber, C., Kupatt, C., & Engelhardt, S. (2020). AntimiR-21 Prevents Myocardial Dysfunction in a Pig Model of Ischemia/Reperfusion Injury. Journal of the American College of Cardiology, 75(15), 1788-1800. https://doi.org/10.1016/j.jacc.2020.02.041

@article{4a89c54fe0874c1bbc5b139ac3bdf53a,

title = "AntimiR-21 Prevents Myocardial Dysfunction in a Pig Model of Ischemia/Reperfusion Injury",

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. (C) 2020 The Authors. Published by Elsevier on behalf of the American College of Cardiology Foundation.",

keywords = "cardiac disease, fibrosis, microRNA, miR-21, porcine model of heart failure, MICRORNA-21",

author = "Rabea Hinkel and Deepak Ramanujam and Veronika Kaczmarek and Andrea Howe and Katharina Klett and Christina Beck and Anne Dueck and Thomas Thum and Karl-Ludwig Laugwitz and Lars Maegdefessel and Christian Weber and Christian Kupatt and Stefan Engelhardt",

note = "Funding Information: This work was funded by BMBF Munich Biotech cluster m4 (grant BMBF16GW0019 to Drs. Kupatt and Engelhardt) and the German Center for Cardiovascular Research (DZHK). The authors{\textquoteright} research is supported by Deutsche Forschungsgemeinschaft (grant TRR267 to Drs. Kupatt, Maegdefessel, Dueck, Laugwitz, Weber, and Engelhardt and grant SFB1123 to Drs. Maegdefessel and Weber) and the European Union (ERA-CVD MacroERA [01KL1706] to Dr. Engelhardt). Dr. Weber is a Van de Laar Professor of atherosclerosis. Drs. Thum and Engelhardt have received royalties from a patent on miR-21 licensed by Uni W{\"u}rzburg. Dr. Thum is founder and holds shares of Cardior Pharmaceuticals GmbH. All other authors have reported that they have no relationships relevant to the contents of this paper to disclose. Publisher Copyright: {\textcopyright} 2020 The Authors",

year = "2020",

month = apr,

day = "21",

doi = "10.1016/j.jacc.2020.02.041",

language = "English",

volume = "75",

pages = "1788--1800",

journal = "Journal of the American College of Cardiology",

issn = "0735-1097",

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Hinkel, R, Ramanujam, D, Kaczmarek, V, Howe, A, Klett, K, Beck, C, Dueck, A, Thum, T, Laugwitz, K-L, Maegdefessel, L, Weber, C, Kupatt, C & Engelhardt, S 2020, 'AntimiR-21 Prevents Myocardial Dysfunction in a Pig Model of Ischemia/Reperfusion Injury', Journal of the American College of Cardiology, vol. 75, no. 15, pp. 1788-1800. https://doi.org/10.1016/j.jacc.2020.02.041

TY - JOUR

T1 - AntimiR-21 Prevents Myocardial Dysfunction in a Pig Model of Ischemia/Reperfusion Injury

AU - Hinkel, Rabea

AU - Ramanujam, Deepak

AU - Kaczmarek, Veronika

AU - Howe, Andrea

AU - Klett, Katharina

AU - Beck, Christina

AU - Dueck, Anne

AU - Thum, Thomas

AU - Laugwitz, Karl-Ludwig

AU - Maegdefessel, Lars

AU - Weber, Christian

AU - Kupatt, Christian

AU - Engelhardt, Stefan

N1 - Funding Information: This work was funded by BMBF Munich Biotech cluster m4 (grant BMBF16GW0019 to Drs. Kupatt and Engelhardt) and the German Center for Cardiovascular Research (DZHK). The authors’ research is supported by Deutsche Forschungsgemeinschaft (grant TRR267 to Drs. Kupatt, Maegdefessel, Dueck, Laugwitz, Weber, and Engelhardt and grant SFB1123 to Drs. Maegdefessel and Weber) and the European Union (ERA-CVD MacroERA [01KL1706] to Dr. Engelhardt). Dr. Weber is a Van de Laar Professor of atherosclerosis. Drs. Thum and Engelhardt have received royalties from a patent on miR-21 licensed by Uni Würzburg. Dr. Thum is founder and holds shares of Cardior Pharmaceuticals GmbH. All other authors have reported that they have no relationships relevant to the contents of this paper to disclose. Publisher Copyright: © 2020 The Authors

PY - 2020/4/21

Y1 - 2020/4/21

N2 - 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. (C) 2020 The Authors. Published by Elsevier on behalf of the American College of Cardiology Foundation.

AB - 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. (C) 2020 The Authors. Published by Elsevier on behalf of the American College of Cardiology Foundation.

KW - cardiac disease

KW - fibrosis

KW - microRNA

KW - miR-21

KW - porcine model of heart failure

KW - MICRORNA-21

U2 - 10.1016/j.jacc.2020.02.041

DO - 10.1016/j.jacc.2020.02.041

M3 - Article

C2 - 32299591

SN - 0735-1097

VL - 75

SP - 1788

EP - 1800

JO - Journal of the American College of Cardiology

JF - Journal of the American College of Cardiology

IS - 15

ER -