Figure 2: Prediction and validation of biological targets/mechanism of action (MOA) of antineoplastic compounds.
From: High-resolution phenotypic profiling of natural products-induced effects on the single-cell level
(a) Hierarchical clustering of cytolological profiles composed of 20 core features of a group of 26 anti-neoplastic compounds reveals various distinct functional sub-groups that clearly reflect different molecular targets and MOAs. Colors indicate positive (yellow) or negative (blue) deviation from the mean of untreated control cells (value = 1). Full cytological profiles were used for clustering (left). Cytological profiles showing a reduced set of 20 core markers are displayed for easier visualization of affected processes and a vertical grey bar was used to separate between regulatory (i.e. NFkB, p53, and caspase 9 activation) and other cellular markers. Colors indicate positive (yellow) or negative (blue) deviation from the mean of untreated control cells (value = 1). The dendrogram depicts distances between individual cytological profiles based on Spearman rank correlation. (b) Four candidate natural products and three reference compounds were tested for phosphorylation of H2AX in a cell based assay to assess DNA double strand breaks caused by topoisomerase II poisons (control shows no effects). (c) Detection of phospho-H2AX in HeLa cells treated with vehicle control (non-treated) or with 20 μM of compound 4 (see d) for 6 hours. (d) Structures of selected natural products and reference compounds of the topoisomerase cluster from (a): derivative of 10-hydroxycamptothecin (4), Norsecurinine (5), Dorsmanin E (6), 2-(3-Acetoxy-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]-phenanthren-17-yl)-2-hydroxypropyl acetate (7), (S)-(+)-Camptothecin, beta-Lapachone, and Genistein.
