Evaluating Cellular Pharmacodynamics of Small Molecule Drugs

Example 4

Cell-based using assays using human lens epithelial cell lines SRA 01/04 and B3 were also performed. The cells were exposed to UV or heat and cell viability assessed 24 hours post-exposure by Alamar blue staining. The results are shown in FIGS. 7A and 7B. In another experiment, SRA 01/04 cells were pre-incubated with varying concentrations of compounds two hours prior to UV exposure. Relative protection was measured as percent viability compared to vehicle control 24 hours following UV exposure. Effect of compounds from each of three distinct chemical series (macrocyclics, covalent, and catechol) are shown in FIG. 7B. Mean±SEM of the measurements are shown.

Ectopic expression of AAC-EGFP in B-3 cells shows that AAC forms inclusions that co-localize with p62 (arrows) (FIG. 8A). Automated image analysis of >2500 cells is shown in FIG. 8B. A statistically significant increase in GFP-positive inclusions due to AAC overexpression was observed. Mean±SEM of the measurements and p value (t test) are shown. The results show that high-content screening could be used to evaluate the cellular pharmacodynamic effects of SMDs for measuring cellular aggregates of AAC.