Project description:Hsp70 inhibition affects many signaling pathways. We established how these effects are translated into changes in gene expression. Hsp70 is a promising anti-cancer target, and several inhibitors of Hsp70 have been recently developed. Interest to Hsp70 inhibitors as drug prototypes is, however, somewhat hampered by potential similarity of their physiological effects to effects of already well-developed Hsp90 inhibitors. JG-98 series of inhibitors is unique in its ability to target an allosteric site in the ATPase domain of Hsp70, which disrupts its interaction with a co-chaperone Bag3 and affects a variety of signaling pathways important for cancer development and survival. Here, we used the Broad Institute Connectivity Map platform to evaluate physiological effects of JG-98, and found that these effects are dissimilar from effects of Hsp90 inhibitors, thus justifying further development of this compound series. Further, using gene expression data and ActivSignal IPAD platform, we identified pathways modulated by JG-98. Some of these pathways were affected by JG-98 in Bag3-dependent and some pathways in Bag3-independent manner, indicating multiple mechanisms of JG-98 action. Using pooled shRNA genetic screen, we established gene sets that modulate the response of cancer cells to JG-98. Based on genetic and gene expression information, we developed approaches to predict potent combinations of JG-98 with known drugs. These predictions were validated by demonstrating that proteasome, RNApol II, Akt and RTK inhibitors synergize with anti-cancer effects of JG-98. Overall, in this study we analyze unique effects of Hsp70 inhibitors of JG-98 series on cell physiology and define potential drug combinations for clinical use of these inhibitors.
Project description:The whole proteome analysis of the Pseudomonas sp. FIP_A4 strain in presence and absence of fipronil was conducted to evaluate the differentially expressed enzymes that can play role in fipronil degradation.
