Transcriptional pathway profiling in human T cells reveals a novel functional class of thalidomide analogs. (A) Chemical structures of thalidomide compounds profiled in this study. (B) The transcriptional targeting of thalidomide (100 μM), CPS11, CPS45, CPS49, cc5013 (Revlimid; lenalidomide), cc4001 (rolipram), and cc4047 (Actimid) (10 μM each) were profiled by high-throughput transfection12 using the indicated luciferase-based reporters (columns) stimulated by 16 different combinations of T-cell mitogens (rows; key defines numbers). Concentrations of the mitogens used were 50 ng/mL PMA, 720 ng/mL Ion, 1:1000 dilution of anti-CD3 antibody, and 1:1000 dilution of anti-CD28 monoclonal antibody. The transcriptional output was analyzed by hierarchical clustering applying a Euclidian distance metric. Color intensities from green to red are based on percentage of maximum stimulation. (C) PCA shows a separation of CPS45, CPS49, and CPS11 from the other thalidomide compounds along PC1. (D) Percentage variation in the transcriptional data captured by the different principal components. x-axis denotes the PCs; y-axis indicates the amount of variance captured by each PC (black), with cumulative variance shown in blue.
