Slice of Life: Porcine Kidney Slices for Testing Antifibrotic Drugs in a Transplant Setting

Abstract

Circulatory death donor (DCD) kidneys are increasingly used to enlarge the donor pool. These kidneys undergo ischemia-reperfusion injury, frequently leading to renal fibrosis. Transforming growth factor beta 1 (TGF-β1) and matrix metalloproteases have been identified as central mediators of fibrosis and inhibition of these targets could attenuate fibrosis. We studied whether galunisertib, doxycycline, taurine, and febuxostat alleviated fibrosis in precision-cut kidney slices (PCKS). PCKS were prepared from porcine kidneys that were exposed to 30 min of warm ischemia followed by 3 h of oxygenated hypothermic machine perfusion. We subsequently incubated PCKS for 48 h at 37 °C with the described compounds. To further elucidate the antifibrotic effects of galunisertib, we cultured PCKS with TGF-β1. We first screened the effects of the compounds without TGF-β1. Most significant effects were observed for galunisertib which lowered the expression of ACTA2, TGFB1, FN2, and SERPINE1. We then investigated the effects of galunisertib in fibrotic PCKS incubated with TGF-β1. TGF-β1 significantly increased expression of TGFB1, FN1, SERPINE1, and SERPINH1. Galunisertib, however, attenuated the expression of all fibrosis-related genes. Galunisertib appears to be a promising antifibrotic compound requiring further research in a preclinical model and may ultimately be administered during machine perfusion as an antifibrotic treatment in a transplant setting.

Targeted delivery of galunisertib using machine perfusion reduces fibrogenesis in an integrated ex vivo renal transplant and fibrogenesis model

“…(LeBleu et al, 2013), and secretion of the collagen chaperone HSP47 (Razzaque et al, 2005). Similar effects on gene expression of fibrosis markers were observed in previous studies using human, porcine and murine tissue slices (Bigaeva, Gore, & Mutsaers, 2020;Bigaeva, Puerta Cavanzo, et al, 2020;van Leeuwen et al, 2023), although the kidneys described in these studies were not subjected to normothermic machine perfusion.…”

“…Only one kidney (both left and right kidneys) from each pig was used and randomized into experimental groups. The sample size of n = 8 per experimental group was determined based on preliminary experiments (van Leeuwen et al, 2023). For each analysis requiring tissues, three tissue slices were incubated per kidney but pooled for the analysis.…”