Establishment of in vitro nephrotoxicity assessment for antisense oligonucleotides using human renal proximal tubule epithelial cells

抄録

Antisense oligonucleotides (ASOs) induced renal proximal tubule injury is one of the major reasons for drug candidate attrition. There is a need to develop robust in vitro screening assays to identify nephrotoxic ASOs before they reach clinical trials. The purpose of this study is to confirm an in vitro profile and set up screening assays using human primary renal proximal tubule epithelial cells (hRPTEC) that recapitulates the in vivo pre-clinical nephrotoxicity by ASOs.

Effect of ASOs on EGF dependent hRPTEC growth was evaluated to establish in vitro robust assays. A several in-house ASOs that showed in vivo nephrotoxicity in pre-clinical studies, dose dependently inhibited EGF induced hRPTEC growth for 9 days treatments. The data were analyzed for evaluation on relevant safety for in vitro and in vivo correlation. Some nephrotoxic potential ASOs were further investigated on primary human proximal tubule cell (PTC) monolayers using a panel of FDA-approved biomarkers (KIM-1, NGAL, clusterin). These ASOs were treated for 3 days in the absence and presence of Receptor Associated Protein (RAP), an inhibitor of the proposed mechanism of ASOs entry into the cells. RAP partially reduced induction of biomarker production by ASOs.

In summary, our studies suggested that in vitro EGF dependent hRPTEC growth and PTC monolayers models were useful tools for safety evaluation before in vivo pre-clinical nephrotoxicity assessment in early drug discovery process.