Delivery and expression of mRNA therapeutics during ex vivo kidney perfusion: A feasibility study in porcine and human donor kidneys

Ischemia–reperfusion injury (IRI) is a major cause of graft dysfunction in donor kidneys. Normothermic machine perfusion (NMP) offers a clinically relevant ex vivo platform for targeted intervention during organ preservation, while minimizing systemic exposure. Here, we investigated lipid nanoparticle (LNP)-mediated mRNA delivery during NMP as a strategy to enable immediate post-transplant therapeutic activity. Porcine and human kidneys were perfused at 37 °C for 6–12 h with an erythrocyte-based perfusate, and LNP-encapsulated mCherry or human erythropoietin (hEPO) mRNA was administered via the renal artery. Intravascular LNP delivery resulted in widespread renal parenchymal uptake of reporter mRNA without evidence of vascular obstruction or histological injury. hEPO protein became detectable in the perfusate approximately 2 h after infusion and continued to increase throughout perfusion, reaching biologically relevant concentrations. LNPs incorporating PEG-lipid TPGS (D-α-tocopherol polyethylene glycol 1000 succinate) achieved significantly higher mRNA delivery efficiency than conventional DSPE-PEG2000 formulations. The findings provide a foundation for the use of LNPs and mRNA therapeutics during organ machine perfusion as a platform for precision medicine in the field of kidney transplantation.