Abstract P6-04-07: Significance and therapeutic potential of PELP1-mTOR axis in breast cancer progression and therapy resistance
Authors: Gonugunta VK , Cortez V , Sareddy GR , Roy SS , Zhang H , Tekmal RR , Vadlamudi RK 
Published: August 2014

Abstract
Proline, Glutamic-acid and Leucine-rich Protein 1 (PELP1) is a proto-oncogene that modulates ER signaling by functioning as an ER-coregulator. Emerging studies demonstrated that in a subset of breast tumors, PELP1 is predominantly localized in the cytoplasm and that PELP1 participates in extranuclear signaling by facilitating ER interactions with Src, PI3K, and AKT. PELP1 expression is upregulated in breast cancer, its deregulation contributes to therapy resistance, and PELP1 is a prognostic marker of poor survival. However, the mechanism by which PELP1 extranuclear actions contributes to cancer progression and therapy resistance remains unknown. We have recently discovered that PELP1 has the potential to interact with mammalian target of rapamycin (mTOR), a serine/threonine kinase that forms two distinct complexes called mTORC1 (containing Raptor and PRAS40) and mTORC2 (containing Rictor and Protor). The objective of this application is to test whether crosstalk occurs between mTOR and PELP1 signaling axis and to test whether mTOR targeting drugs can be used to target PELP1 oncogenic functions. We have used breast cancer cells with PELP1 overexpression (MCF7-PELP1, ZR75-PELP1, T47D-PELP1) or PELP1 down regulation (MCF7-PELP1shRNA, ZR75-PELP1shRNA) along with controls to study the role of PELP1 in the regulation of mTOR axis. PELP1 knockdown significantly reduced downstream mTOR signaling components as analyzed by Western analysis using phospho-S6K, -4EBP1, -mTOR and -Akt, antibodies. Overexpression of PELP1 activated mTOR signaling components. Using immunoprecipitation, we have demonstrated that PELP1 interacts with mTOR. Further immunopreciptation analysis using Rictor and Raptor specific antibodies revealed that PELP1 associates with both TORC1 and TORC2 complexes. Using PELP1WT and PELP1cyto (that predominantly localizes in the cytoplasm), we have demonstrated the differential activation of mTOR signaling components: PELP1WT activated both TORC1 and TORC2 pathways, while PELP1cyto uniquely activated TORC2. mTOR targeting drugs (Rapamycin or AZD8055) showed a significant effect on the in vitro proliferation of PELP1 model cells. AZD8055 is more potent in reducing PELP1 driven tumor growth in vivo compared to rapamycin. Immunohistochemical studies on xenografts derived from MCF7, MCF7-PELP1WT and MCF7-PELP1cyto models demonstrated that PELP1 signaling modulates mTOR signaling in vivo and inhibition of mTOR signaling rendered PELP1 driven tumors to be highly sensitive to therapeutic inhibition. Further, mTOR inhibitors sensitized tamoxifen therapy resistant PELP1cyto model cells to hormonal therapy. IHC analysis of mammary glands and mammary tumors from PELP1Tg mice revealed deregulation of mTOR signaling components with excessive activation of S6K and 4EBP1. Using breast tumor tissue arrays (n = 100), we found significant correlation of PELP1 cytosolic localization with mTOR signaling. Collectively, the experimental results from these studies identified PELP1-mTOR axis as a novel component of PELP1 oncogenic functions and suggests, mTOR inhibitor(s) will be effective chemotherapeutic agents for down regulating PELP1 oncogenic functions and for blocking PELP1-mediated therapy resistance.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-04-07.