MicroRNAs in extracellular vesicles released from epicardial adipose tissue promote arrhythmogenic conduction slowing

Auriane C Ernault; Rosan de Winter; Benedetta Fabrizi; Jillian W P Bracht; Chi Hau; Shirley C M van Amersfoorth; Eva R Meulendijks; Anke J Tijsen; Lucía Cócera Ortega; Ingeborg van der Made; Aleksandra Gasecka; Antoine H Driessen; Rienk Nieuwland; Bastiaan J Boukens; Edwin van der Pol; Joris R de Groot; Ruben Coronel

doi:10.1016/j.hroo.2023.10.007

MicroRNAs in extracellular vesicles released from epicardial adipose tissue promote arrhythmogenic conduction slowing

Heart Rhythm O2. 2023 Nov 2;4(12):805-814. doi: 10.1016/j.hroo.2023.10.007. eCollection 2023 Dec.

Authors

Auriane C Ernault^{1

2}, Rosan de Winter^{1

2}, Benedetta Fabrizi^{1

2}, Jillian W P Bracht^{3

4}, Chi Hau^{3

4}, Shirley C M van Amersfoorth^{1

2}, Eva R Meulendijks^{1

2}, Anke J Tijsen^{1

2}, Lucía Cócera Ortega^{1

2}, Ingeborg van der Made^{1

2}, Aleksandra Gasecka^{3

4

5}, Antoine H Driessen^{1

2}, Rienk Nieuwland^{3

4}, Bastiaan J Boukens^{2

6}, Edwin van der Pol^{3

4

7}, Joris R de Groot^{1

2}, Ruben Coronel^{1

2}

Affiliations

¹ Department of Clinical and Experimental Cardiology, Amsterdam UMC, location University of Amsterdam, Amsterdam, the Netherlands.
² Heart Failure and Arrhythmias, Amsterdam Cardiovascular Sciences, Amsterdam, the Netherlands.
³ Laboratory of Experimental Clinical Chemistry, Amsterdam UMC, location AMC, Amsterdam, the Netherlands.
⁴ Vesicle Observation Center, Amsterdam UMC, location AMC, Amsterdam, the Netherlands.
⁵ Department of Cardiology, Medical University of Warsaw, Warsaw, Poland.
⁶ Department of Medical Biology, Amsterdam UMC, location University of Amsterdam, Amsterdam, the Netherlands.
⁷ Biomedical Engineering and Physics, Amsterdam UMC, location AMC, Amsterdam, the Netherlands.

Abstract

Background: Patients with excess epicardial adipose tissue (EAT) are at increased risk of developing cardiac arrhythmias. EAT promotes arrhythmias by depolarizing the resting membrane of cardiomyocytes, which slows down conduction and facilitates re-entrant arrhythmias. We hypothesized that EAT slows conduction by secreting extracellular vesicles (EVs) and their microRNA (miRNA) cargo.

Objective: We aimed to determine the role of EAT-derived EVs and their miRNA cargo in conduction slowing.

Methods: EAT and subcutaneous adipose tissue (SAT) were collected from patients with atrial fibrillation. Adipose tissue explants were incubated in culture medium and secretome was collected. The numbers of EVs in the EAT and SAT secretome were measured by calibrated flow cytometry. EVs in the EAT secretome were isolated by size exclusion chromatography and miRNAs were sequenced. Pathway analysis was performed to predict candidates involved in cardiac electrophysiology. The candidates were validated in the EAT and SAT by quantitative real-time polymerase chain reaction. Finally, miRNA candidates were overexpressed in neonatal rat ventricular myocytes.

Results: The EV concentration was higher in the EAT secretome than in the SAT and control secretomes. miRNA sequencing of EAT-derived EVs detected a total of 824 miRNAs. Pathway analysis led to the identification of 7 miRNAs potentially involved in regulation of cardiac resting membrane potential. Validation of those miRNA candidates showed that they were all expressed in EAT, and that miR-1-3p and miR-133a-3p were upregulated in EAT in comparison with SAT. Overexpression of miR-1-3p and miR-133a-3p in neonatal rat ventricular myocytes led to conduction slowing and reduced Kcnj2 and Kcnj12 expression.

Conclusion: miR-1-3p and miR-133a-3p are potential mediators of EAT arrhythmogenicity.

Keywords: Adiposity; Arrhythmias; Atrial fibrillation; Epicardial adipose tissue; Obesity; miRNA.