O3 Ex vivo normothermic machine perfusion facilitates gymnotic delivery of RNA interference therapeutics in donor kidneys

Abstract Introduction Normothermic machine perfusion (NMP) of donor kidneys prior to transplantation provides a platform for delivery novel therapeutics optimize organ quality. This includes RNA interference (RNAi) e.g. antisense oligonucleotides (ASO) that block detrimental microRNAs. The intracellular kinetics RNAi are crucial their pharmacological effect, however, it remains poorly understood. NMP an ideal investigate this further. Method During NMP, human (n = 12) were treated 6 hours with fluorescently-labelled ASO designed microRNA-24-3p activity. Biopsies taken at 0, 2, 4, and hours. Kidney sections stained antibodies against early endosomes (Rab5), late (Rab7), RNA-induced silencing complexes (GW182) lysosomes (LAMP2). Confocal microscopy images obtained co-localisation quantified using Hugyens™ software following batch deconvolution. global transcriptomic impact therapy was also assessed sequencing. Result Following 2 primarily found in tubular epithelial cells. Co-localisation studies revealed uptake via endocytosis endosomal sorting occurring during NMP. followed by cytoplasmic escape GW182 proteins. pattern not seen scrambled sequence or cold controls. RNAseq analysis decrease inflammatory pathways upregulation targets. Discussion is the first study demonstrate facilitates gymnotic directly into RISC, whereby, blocks microRNA-mediated mRNA increases bioavailability protective highlights capacity re-programme gene expression therapeutics. Take-home Message Ex vivo normothermic unique window opportunity when we can deliver therapies improve quality organ. Novel genetic protect ischemia reperfusion injury could potentially increase utilisation post-transplant outcomes many patients on kidney transplant waiting list.