Ex-vivo mRNA therapy to predict kidney vitality during transplantation

TY - CONF

T1 - Ex-vivo mRNA therapy to predict kidney vitality during transplantation

AU - Song, Ping

AU - Staulund, Jesper

AU - Qi, Haiyun

AU - Thrane, Lars

AU - Pedersen, Michael

AU - Eijken, Marco

AU - Jespersen, Bente

AU - Kjems, Jørgen

PY - 2022/10/11

Y1 - 2022/10/11

N2 - 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. 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. 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.We envision that the strategy can potentiate the application to various diseases and target tissues.

AB - 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. 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. 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.We envision that the strategy can potentiate the application to various diseases and target tissues.

KW - Kidney vitality prediction

KW - RNA delivery

KW - Lipid nanoparticles

M3 - Poster

T2 - Danish RNA Society's 7th Annual Meeting

Y2 - 11 October 2022 through 11 October 2022

ER -