P30 Target discovery for prevention of hypertrophic scars using an annotated compound library

Abstract Introduction and aims It is estimated that 100 million people in the developed world develop dermal scarring as the result of a burn injury, trauma or surgery every year. It affects not only the mental wellbeing of patients, but also physical effects such as itching, pain and loss of mobility in affected areas. Current treatment options are limited in their effectiveness and there are currently no approved medications, highlighting an unmet need. The aim for this project was to screen a fully annotated small molecule library from AstraZeneca to discover possible compounds for prevention of hypertrophic scars. Methods A total of 3500 compounds with known pharmacology and targets were screened in triplicate in a 96-well assay format using human primary dermal fibroblasts which were transformed to myofibroblasts using transforming growth factor-β1. α-Smooth muscle actin expression was quantified using the in-cell enzyme-linked immunosorbent assay (ICE) method, with cell viability measured using DRAQ5 nuclear staining. Hit compounds were selected based on their potency and selectivity to targets. Presence of targets in human dermal fibroblasts was confirmed by RNA sequencing (RNAseq). In total, 33 compounds met hit criteria (more than 50% inhibition at 0.1 µmol L−1, with ≥ 80% viability across all compound concentrations), with 32 compounds added from the RNAseq data. Eleven-point concentration–response curves (CRC) were constructed for all 65 compounds in triplicate and in 3 different cell lines, generating IC50 data. Results Overall, 39 compounds generated CRCs, allowing for IC50 data, and therefore pIC50 to be generated. pIC50 values ranged from 8.72 to 3.47, with 26 compounds showing no pharmacological activity. Conclusions Of the 3500 compounds screened, 39 compounds generated IC50 data while maintaining hit criteria, highlighting possible future compounds to be repurposed for prevention of hypertrophic scars. Next steps include investigation of near-neighbour compounds.