Abstract 1038: Inhibition of mTOR enhances the activity of HSP90 inhibitors in part through cessation of heat shock protein synthesis.

Abstract Background: Heat shock protein 90 (HSP90) is a molecular chaperone required for the stability of hundreds of client proteins, many of which promote tumor growth. While HSP90 inactivation abrogates multiple oncogenic pathways, it subsequently triggers a heat shock response that may limit its full cytotoxic potential. To overcome this limitation, we sought to identify a clinically feasible method to block heat shock protein synthesis induced by the HSP90 inhibitor ganetespib. Results: An immunoassay was developed to screen a library of >300 late stage or approved drugs for their ability to block the upregulation of HSP70 subsequent to HSP90 inhibition. A number of effective agents were identified, with the most prevalent and effective being those that target PI3K/mTOR signaling. Validating this result, mTOR inhibitors diminished ganetespib induced HSP70 and HSP90 protein levels in multiple cancer cell lines. To determine the mechanism of HSP regulation by mTOR inhibitors the expression of >80 heat shock related genes was analyzed. Inhibitors of mTOR had no effect on HSP RNA levels but suppressed the upregulation of HSP genes induced by ganetespib. Heat shock factor 1 (HSF1) is the major transcriptional regulator of HSP's. Under conditions of stress or HSP90 inhibition, HSF1 is released from repressive cytoplasmic chaperone complexes and translocates into the nucleus to initiate heat shock gene transcription. Nuclear HSF1 levels increased within 1 h of ganetespib exposure but were dramatically reduced in the presence of an mTOR inhibitor. A major function of mTOR is regulating protein synthesis through activation of the cap-binding protein EIF4E, therefore we investigated whether inhibition of protein synthesis would affect HSF1 nuclear accumulation. Cycloheximide or an EIF4E inhibitor reduced ganetespib-mediated HSF1 entry into the nucleus suggesting that inhibition of mTOR can prevent HSP induction by blocking protein synthesis. Combinations of ganetespib with mTOR or dual mTOR/PI3K inhibitors resulted in a significant increase in anticancer activity compared to monotherapy in multiple in vitro and in vivo cancer models. Conclusions: Inhibition of mTOR counteracts ganetespib induced upregulation of HSP's by blocking translocation of HSF1 into the nucleus, an effect potentially mediated by a decrease in mTOR driven protein synthesis. Attenuation of heat shock response may contribute to the synergy observed for ganetespib and PI3K/mTOR inhibitors in preclinical cancer models. Targeting mTOR signaling thus represents a promising, clinically feasible approach to maximize the therapeutic potential of ganetespib. Citation Format: Suqin He, Jaime Acquaviva, Julie C. Friedland, Jim Sang, Donald L. Smith, Manuel Sequeira, Chaohua Zhang, David A. Proia. Inhibition of mTOR enhances the activity of HSP90 inhibitors in part through cessation of heat shock protein synthesis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1038. doi:10.1158/1538-7445.AM2013-1038