Figure 5

Safety Evaluation of ASO chemistry in PT-MPS. (a) Schematic of the PT-MPS with a phase contrast image of a confluent PTEC tubule. (b) Representative images of tubules stained for HO-1 (red) by immunocytochemistry after 5 days of treatment with 1 µM HPV PMO or 1 µM DSP PSO (scale = 100 µm). Cells display a predominately perinuclear and cytoplasmic HO-1 signal. (c) Quantification of HO-1 fluorescence intensity from tubules depicted in B. Expression and localization of HO-1 is not significantly affected by HPV PMO or DSP PSO treatment. (d) Quantification of levels of KIM-1 in device effluent over time by mesoscale chemiluminescent assay after treatment with 1 µM HPV PMO or 1 µM DSP PSO. There is a significant reduction in KIM-1 secretion with PMO treatment compared to control (p = 0.043, ANOVA with Tukey’s adjustment) and PSO groups (p = 0.01, ANOVA with Tukey’s adjustment) (e) Quantification of levels of a panel of renal disease- and injury-associated miRNAs by qPCR after treatment with 10 µM HPV PMO or 10 µM DSP PSO for 44 h. Data are presented as the delta-deltaC_T fold change in miRNA levels relative to control. Secretion of miR-30e-5p (p = 0.024, ANONVA, with Tukey’s adjustment) was significantly increased by DSP PSO treatment. (f) Quantification of levels of KIM-1 in device effluent over time by ELISA after treatment with 10 µM HPV PMO or 10 µM HPV PSO. There was no time- or dose- dependent effect of HPV PMO, DSP PSO, or HPV PSO treatment on the secretion of KIM-1. For all quantitative panels (C-F), data are presented as mean ± s.d with black circles representing the value for each individual tubule tested.