Figure 4.

High content drug screen of targeted anti-cancer library to profile their effects on the metastatic behavior of MDA-MB-231 tumor cells. (A) Ordered bar chart distribution showing the drug screen data using metastasis-on-chip model. The intravasation and invasion were normalized to controls treated with vehicle (DMSO). Compound inhibitor of tumor intravasation were indicated below the dotted line (hit threshold was calculated as follows: mean_(vehicle) - 4x standard deviation_(vehicle). Drug library was screen at final concentration of 5 μM. (B) Representative confocal Z-stack images for the negative control (DMSO, 0.02%) and the positive control (ROCK inhibitor compound “GSK429286”, 5 μM). Scale bar = 250 μm. (C) Compound hit list that reduce cancer breast cell intravasation with their molecular targets. In red, drugs approved clinically as monotherapies for triple-negative breast cancer. (D) Uniform manifold approximation (UMAP) analysis comparing phenotypic profile of drug library based upon identified phenotypic groupings. Drugs showed similar phenotypes are gathered in one cluster on the plot. Data point color indicates which cluster each molecule belongs to. (E) heatmap showing the phenotypic features of each clusters (F) Enrichment analysis of the drug molecular targets was performed by Fisher exact test. Chart showing the molecular targets were significantly enriched in each cluster (p<0.05). Red color intensity is proportional to the significance of Fisher exact test.