Exploring ex vivo modulation of fibrosis in discarded human donor kidneys

Pamplona, C. C.; Bergada, C. B.; Jaynes, C. L.; Hamelink, T. L.; Lantinga, V. A.; Ogurlu, B.; Ottens, P. J.; van Hateren, K.; Hillebrands, J.-L.; Olinga, P.; Touw, D. J.; Leuvenink, H. G. D.; Moers, C.; van Leeuwen, L. L.

doi:10.1101/2025.07.31.667749

bioRxiv · 10.1101/2025.07.31.667749

Exploring ex vivo modulation of fibrosis in discarded human donor kidneys

Pamplona, C. C.·Bergada, C. B.·Jaynes, C. L.·Hamelink, T. L.·Lantinga, V. A.·Ogurlu, B.·Ottens, P. J.·van Hateren, K.·Hillebrands, J.-L.·Olinga, P.·Touw, D. J.·Leuvenink, H. G. D.·Moers, C.·van Leeuwen, L. L.

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Abstract

IntroductionEarly-onset fibrosis limits kidney transplant success. Normothermic machine perfusion (NMP) offers a platform for targeted drug delivery directly to isolated organs, minimizing systemic effects. This study evaluated the long-term anti-fibrotic efficacy and safety of galunisertib in discarded human kidneys perfused ex vivo. MethodsTwelve discarded human kidneys underwent 4 hours of oxygenated hypothermic perfusion followed by 6 hours of NMP with galunisertib or vehicle (n=6). Precision-cut kidney slices (PCKS) were then cultured for 48 hours with either continued or discontinued galunisertib exposure. Endpoints included fibrosis-related mRNA expression and pharmacokinetics. ResultsGalunisertib did not negatively affect renal function during NMP. Continued exposure in PCKS significantly attenuated fibrosis-related mRNA expression, including SERPINE1 (p=0.0046), TGF-{beta} (p=0.0168), FN1 (p=0.0269) and ACTA2 (p=0.0014) after 48 hours. The discontinuation of treatment did not exhibit the same anti-fibrotic effects. ConclusionGalunisertib was safely administered during NMP and steadily excreted via the urine. NMP showed to be a promising platform for safe targeted anti-fibrotic therapy delivery, offering potential to improve graft quality. When treatment was sustained, galunisertib induced a modest reduction in fibrosis-related mRNA expression over 48 hours of tissue incubation. Further studies are needed to optimize delivery strategies and evaluate the impact of prolonged therapeutic exposure.

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Pamplona, C. C., Bergada, C. B., Jaynes, C. L., Hamelink, T. L., Lantinga, V. A., Ogurlu, B., Ottens, P. J., van Hateren, K., Hillebrands, J.-L., Olinga, P., Touw, D. J., Leuvenink, H. G. D., Moers, C., van Leeuwen, L. L.. 2025-08-02. Exploring ex vivo modulation of fibrosis in discarded human donor kidneys. https://doi.org/10.1101/2025.07.31.667749

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