Recellularized native kidney scaffolds as a novel tool in nephrotoxicity screening

Abstract

Drug-induced kidney injury (DIKI) in medicinal compound development accounts for over 20% of clinical trials failure and involves damage to different nephron segments, mostly the proximal tubule. Yet, currently applied cell models fail to reliably predict nephrotoxicity nor are easy to establish. Here, we developed a novel three-dimensional (3D) nephrotoxicity platform based on decellularized rat kidneys scaffolds (DS) recellularized with conditionally immortalized human renal proximal tubule epithelial cells overexpressing the organic anion transporter 1 (ciPTEC-OAT1). A 5-days sodium dodecyl sulfate (SDS)-based decellularization protocol was used to generate DS , of which 100 um slices were cut and used for cell seeding. After 8 days of culturing, recellularized scaffolds (RS) demonstrated 3D tubules formation along with tubular epithelial characteristics, including drug transporter funcion. Exposure of RS to cisplatin (CDDP), tenofovir (TFV) or cyclosporin A (CsA) as prototypical nephrotoxic drugs, revealed concentration-dependent reduction in cell viability, as assessed by PrestoBlue and Live/Dead staining assays. CDDP, TFV and CsA TC50-values were 108 ± 1 and 12 ± 1, 212 ± 1 and 97 ± 1, 129 ± 2 and 294 ± 2 μM in 2D and 3D cultures, respectively. This was most likely due to specific uptake of CDDP by the organic cation transporter 2 (OCT2), TFV through organic anion transporter 1 (OAT1) and CsA competing for P-glycoprotein-mediated efflux. As compared to 2D cultures, RS showed an increased sensitivity to cisplatin and tenofovir toxicity after 24h exposure (9 and 2.2 fold, respectively). In conclusion, we developed a physiologically-relevant 3D nephrotoxicity screening platform that could potentially be suitable as a novel alternative in drug development.