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Updated: Jan 7, 2026
A High-Fidelity Porcine Model of Orthotopic Heart Transplantation Following Donation after Circulatory Death
Published on: June 6, 2025
Left ventricular transcriptome and extracellular vesicle-derived miRNAs in porcine donation after circulatory death
Fulin Wang1, Eliana Lucchinettim2, Phing-How Lou3
1Department of Pharmacology, University of Alberta, Edmonton, Canada; Department of Anesthesiology and Pain Medicine, University of Alberta, Edmonton, Canada.
Insights
A novel cardioprotective treatment improves donation after circulatory death (DCD) heart function by promoting adaptive gene expression and beneficial extracellular vesicle (EV)-derived miRNA release during ex situ heart perfusion (ESHP). This mitigates injury and maladaptive remodeling in DCD hearts.
Area of Science:
- Cardiology
- Transplantation Immunology
- Molecular Biology
Background:
- Donation after circulatory death (DCD) hearts suffer from warm ischemia-reperfusion injury, limiting clinical utility.
- A combination therapy of Intralipid, sevoflurane, and remifentanil previously improved porcine DCD heart function during ex situ heart perfusion (ESHP).
- This study investigates the molecular mechanisms, including transcriptional and extracellular vesicle (EV)-derived miRNA responses, underlying this cardioprotective effect.
Purpose of the Study:
- To elucidate the transcriptomic and EV-miRNA profiles of DCD hearts treated with a cardioprotective agent during prolonged ESHP.
- To understand how the treatment mitigates ischemia-reperfusion injury and promotes adaptive cellular responses.
Main Methods:
- Porcine DCD hearts underwent 6-hour ESHP, with or without the cardioprotective treatment.
- mRNA sequencing of left ventricular tissue and miRNA sequencing of EVs from perfusate were performed.
- Differential gene expression and pathway analysis were conducted using DESeq2 and gene set enrichment analysis.
Main Results:
- DCD hearts showed significant transcriptomic alterations compared to healthy controls, including downregulated protein synthesis and upregulated inflammatory pathways.
- The cardioprotective treatment upregulated pathways related to lipolysis, beta-oxidation, and DNA repair.
- EV-derived miRNAs reflected treatment effects, with pro-survival miRNAs in treated hearts and injury-associated miRNAs in untreated hearts.
Conclusions:
- Cardioprotection during prolonged ESHP induces adaptive transcriptional changes in DCD hearts.
- The treatment promotes beneficial EV-derived miRNA release, counteracting maladaptive remodeling.
- This strategy holds promise for improving the viability and clinical application of DCD hearts.
Background:
Donation after circulatory death (DCD) hearts have limited clinical use due to warm ischemia-reperfusion injury, which impairs graft quality. We previously demonstrated that a cardioprotective treatment combining 1% Intralipid, 2% (v/v) sevoflurane, and 3 nM remifentanil improved the function and viability of porcine DCD hearts during ex situ heart perfusion (ESHP). Here, we used transcriptomics to investigate how the cardioprotective treatment and prolonged ESHP elicit transcriptional and extracellular vesicle (EV)-derived miRNA responses.
Methods:
Porcine DCD hearts were perfused with (treated, n=5) or without (untreated, n=5) the cardioprotective treatment for 6 h. Next generation miRNA sequencing was performed on EVs isolated from perfusate at 1 h reperfusion, and mRNA sequencing was carried out on left ventricular tissue collected at the end of reperfusion, with healthy hearts serving as control. RNA sequencing data was analyzed using gene set enrichment analysis and the R package DESeq2.
Results:
Compared with healthy hearts, 5'483 transcripts were differentially expressed in perfused DCD hearts, including downregulation of protein synthesis, metabolic, and DNA repair pathways, and upregulation of inflammatory and remodeling pathways. Between treated and untreated DCD hearts, 43 transcripts differed significantly. Cardioprotection upregulated gene sets associated with lipolysis, beta-oxidation, and DNA repair, whereas untreated hearts exhibited enrichment of lipid accumulation, oxidative stress, and maladaptive remodeling pathways. EV-derived miRNAs released during reperfusion corroborated these results, with treated hearts releasing pro-survival miRNAs and untreated hearts releasing injury-associated miRNAs.
Conclusions:
Cardioprotection during prolonged ESHP promotes adaptive transcriptional responses and beneficial EV-derived miRNA release, mitigating maladaptive remodeling in DCD hearts.
