Glomerular filtration drug injury: In vitro evaluation of functional and morphological podocyte perturbations

Abstract

The kidney is an organ that plays a major role in the excretion of numerous compounds such as drugs and chemicals. However, a great number of pharmacological molecules are nephrotoxic, affecting the efficiency of the treatment and increasing morbidity or mortality. Focusing on glomerular filtration, we propose in this study a simple and reproducible in vitro human model that is able to bring to light a functional podocyte injury, correlated with morphologic/phenotypic changes after drug exposure. This model was used for the evaluation of paracellular permeability of FITC-dextran molecules as well as FITC-BSA after different treatments. Puromycin aminonucleoside and adriamycin, compounds known to induce proteinuria in vivo and that serve here as positive nephrotoxic drug controls, were able to induce an important increase in fluorescent probe passage through the cell monolayer. Different molecules were then evaluated for their potential effect on podocyte filtration. Our results demonstrated that a drug effect could be time dependent, stable or scalable and relatively specific. Immunofluorescence studies indicated that these functional perturbations were due to cytoskeletal perturbations, monolayer disassembly or could be correlated with a decrease in nephrin expression and/or ZO-1 relocation. Taken together, our results demonstrated that this in vitro human model represents an interesting tool for the screening of the renal toxicity of drugs.

Oestrogen inhibits PTPRO to prevent the apoptosis of renal podocytes

“…Podocytes attach to the outer side of the glomerular basement membrane to form the glomerular hemofiltration barrier between vascular endothelial cells and the glomerular basement membrane (19)(20)(21). Therefore, podocyte apoptosis will increase glomerular basement membrane permeability, inducing proteinuria (20,22,23), which is one of the risk factors for the pathogenesis of childhood nephrotic syndrome. However, little is known about the mechanism of podocyte apoptosis.…”

Molecular and functional characterization of urine‐derived podocytes from patients with Alport syndrome

“…In our co-culture model, we preferred to avoid the addition of an exogenous collagen IV coating, as this GBM component is defective in AS [42]. In this system, we measured the passage of BSA-FITC from the lower (GEC compartment) to the upper (podocyte compartment) side, which is driven by osmotic forces and is halted by the presence of cellular elements [42,43]. Similar to the defective glomerular permeability in AS patients [1], AS podocyte-GEC co-cultures showed significantly increased filtration of albumin compared with the control co-cultures.…”

Effects of heparin and derivatives on podocytes: An in vitro functional and morphological evaluation

“…Human podocytes, kindly provided by Saleem et al (2002), were routinely cultured in RPMI-1640 medium containing 10% foetal bovine serum, 1% insulin-transferrin-selenium-A supplement, and 1% penicillin-streptomycin solution as previously described (Delézay et al, 2017). The cells were cultured at 33°C with 95% air and 5% CO 2 in 75 cm 2 flasks (BD Falcon; Le Pont de Claix, France).…”

“…Delézay et al, 2017), undifferentiated podocytes are smaller than differentiated ones that are spreading on the culture support. The morphological changes observed with UFHtreated differentiated cells prompted us to analyse the effect of this treatment on cellular dedifferentiation.…”