Comparison of the Impact of VRP-034 and Polymyxin B upon markers of Kidney Injury in Human Proximal Tubule Monolayers In Vitro

In this study, we assessed the impact of commercially available polymyxin B against VRP-034 (novel formulation of polymyxin B) using a validated in vitro human renal proximal tubule model.

Freshly isolated primary proximal tubule cells (PTCs) were cultured in Transwell plates and treated with various concentrations of the formulations for up to 48 h. The functional expression of megalin–cubilin receptors in PTC monolayers was validated using FITC-conjugated albumin up-take assays. Polymyxin B and VRP-034 were evaluated at six concentrations (0.3, 1, 3, 10, 30, and 60 µM), and nephrotoxicity was assessed through measurements of transepithelial electrical resistance (TEER), intracellular adenosine triphosphate (ATP) levels, lactate dehydrogenase (LDH) release, and novel injury biomarkers [kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL), and clusterin]. Additionally, histological analysis using annexin V apoptosis staining was performed. Our results indicated a significant decrease in TEER with polymyxin B at concentrations ≥10 µM compared to VRP-034. Toxic effects were observed from ATP and LDH release only at concentrations ≥30 µM for both formulations. Furthermore, injury biomarker release was higher with polymyxin B compared to VRP-034, particularly at concentrations ≥10 µM. Histologically, polymyxin B-treated PTCs showed increased apoptosis compared to VRP-034-treated cells. Overall, VRP-034 demonstrated improved tolerance in the aProximate^TM model compared to polymyxin B, suggesting its potential as a safer alternative for renal protection.

Find out more about aProximate^TM model

Our data demonstrate that VRP-034 is less likely to induce hPTC migration into apoptotic nodules in vitro

Reference

Pye, K.; Tasinato, E.; Shuttleworth, S.; Devlin, C.; Brown, C. Comparison of the Impact of VRP-034 and Polymyxin B upon Markers of Kidney Injury in Human Proximal Tubule Monolayers In Vitro. Antibiotics 2024, 13, 530. https://doi.org/10.3390/antibiotics13060530