• Jespersen, Bente
• Pedersen, Michael
• Song, Ping
• Staulund, Jesper
• Kjems, Jørgen
• Qi, Haiyun
• Eijken, Marco
• Thrane, Lars

Abstract

High-risk kidneys have been used for transplantation due to the big gap between supply and demand for kidney transplantation, which sometimes exhibit unexpected prognosis. Presently, it is still a great challenge to predict the function and metabolic activity of the marginal kidneys for rational preselection in the clinic. Here we utilize non-invasive biodegradable nanoparticles as mRNA carriers to delivery and assess ex-vivo kidney viability and metabolic activity, and finally predict the quality of a donor kidney on a cellular basis for transplantation. <br/><br/>The in vitro performance of lipid nanoparticles (LNPs) for mRNA delivery were screened and validated in HKC-8 cells. Particles loaded with self-quenching dye-conjugated oligos or reporter mRNAs were administered through normothermic machine perfusion system and real-time monitored non-invasively by near infrared (NIF) camera. We hypotheses the uptake and translation are active behaviours of live cells, where only metabolically active cells can emit reporter signal. <br/><br/>Data have shown the accumulation of nanoparticles in pig kidneys and successfully detection of fluorescent signals predominantly in cortex by real-time imaging during ex-vivo perfusion. The delivery of reporter mRNA through developed lipid nanoparticle have shown significant mRNA expression both in vitro and ex-vivo in pig kidneys. The correlation studies to distinguish healthy vs damaged kidneys will be further investigated.<br/><br/>We envision that the strategy can potentiate the application to various diseases and target tissues.<br/>

Topics

• nanoparticle
• impedance spectroscopy
• quenching