PhD Scientific Days 2025

Cardiovascular Medicine and Research II.

Identification and In Vitro Validation of Cardioprotective MicroRNAs From a Porcine Model of Myocardial Infarction and Ischemic Conditioning

Name of the presenter

Szabó Márta

Institute/workplace of the presenter

Semmelweis University - Department of Pharmacology and Pharmacotherapy

Authors

Szabó Márta¹, Nagy Regina¹, Gyovai Áron², Faragó Anna², Makkos András², Görbe Anikó², Ferdinandy Péter²

1: 1Department of Pharmacology and Pharmacotherapy, Nagyvárad tér 4., H-1089, Semmelweis University, Budapest, Hungary; 2Center for Pharmacology and Drug Research & Development, Semmelweis University, Budapest, Hungary
2: 1Department of Pharmacology and Pharmacotherapy, Nagyvárad tér 4., H-1089, Semmelweis University, Budapest, Hungary; 2Center for Pharmacology and Drug Research & Development, Semmelweis University, Budapest, Hungary; 3Pharmahungary Group, H-6722 Szeged, Hungary

Text of the abstract

Introduction: Acute myocardial infarction (AMI) is a leading cause of death worldwide, however, effective pharmacological therapy for ischemia-reperfusion (I/R) injury remains an unmet medical need. MicroRNAs (miRNAs) are small, non-coding RNAs with key roles in post-transcriptional regulation and represent promising cardioprotective targets in ischemic heart disease.
Aims: We aimed to identify novel cardioprotective miRNAs and evaluate their effects in simulated I/R models using neonatal rat and human cardiomyocytes.
Methods: In a previous porcine AMI model subjected to ischemic preconditioning (IPreC), postconditioning (IPostC), and remote perconditioning (RIPerC), we identified 220 differentially expressed miRNAs, including 4 consistently upregulated and 10 downregulated across all protocols. Twelve miRNAs with homologous rat sequences were validated and tested in neonatal rat cardiomyocytes (NRCMs) at 25, 50, and 100 nM, identifying two with significant cardioprotective effects. In this study, six miRNAs (PHmiR-SA201X, -SA202X, -SA203X, -SA204X, -SA205X, -SM102X) with no prior link to cardioprotection were selected for further evaluation.
In the first experimental set, NRCMs were transfected with the selected miRNAs at 6.25, 12.5, and 25 nM using DharmaFECT1 transfection reagent (n=6). After 6 h simulated ischemia and 2 h reperfusion (sI/R), cell viability was assessed. In the second set, the same protocol was applied using lower concentrations: 0.75, 1.5, and 3.125 nM (n=5). In the third set, four miRNAs (PHmiR-SA201X, -SA203X, -SA204X, -SA205X) were tested in human cardiomyocytes (AC16) at 1.5, 3.125, 6.25, 12.5, 25, and 50 nM (n=4), followed by 16 h simulated ischemia and 2 h reperfusion.
Results: PHmiR-SA203X antagomiR significantly improved cell viability at 6.25, 12.5, and 25 nM in NRCMs. No significant effect was observed for any candidate in AC16 cells.
Conclusion: PHmiR-SA203X antagomiR provides significant protection against sI/R injury in NRCMs. The absence of efficacy in human cells indicates the need for further model refinement to improve translational relevance.
Funding: This work was supported by the SE250+ Excellence PhD Scholarship (Semmelweis University) and the National Research, Development and Innovation Office of Hungary (RRF-2.3.1-21-2022-00003).