Abstract NT-116: TARGETING UNFOLDED PROTEIN RESPONSE FOR OVARIAN CANCER THERAPY

Abstract

Abstract Standard paclitaxel-platinum-based chemotherapies often lead to relapses and chemoresistant diseases. New therapeutic strategies are urgently needed for improving the clinical outcomes of ovarian caner patients. Our goal is to identify dysfunctional cellular pathways that are critical for tumor progression and drug resistance and to design novel therapeutic interventions that affect these altered cellular functions. The dysregulation of unfolded protein response (UPR) pathway is often found in cancer cells and contributes to cancer cell survival and their resistance to stress caused by chemotherapies, hypoxia, and nutrition deprivation. The objective of our research is to develop new therapeutic agents to target UPR in ovarian cancer cells in order to overcome chemoresistance. Sulfonamides (SFs) have been used to synthesize antibacterial drugs. We have recently discovered a family of new SFs with anticancer activity. Based on preliminary study, we hypothesize that these new SFs induce apoptosis in ovarian cancer cells through targeting UPR. Using one of the SFs, namely SF-Y3, we compared its effects on epithelial ovarian cancer (EOC) cell lines and immortalized normal fallopian tube (FT) cell lines by performing luminescent CellTiter assay. Phospho-S6 ribosomal protein (P-S6) staining and Annexin V-FITC/PI staining assays were performed to evaluate the effects of SF-Y3 on cell health. Human transcriptome array (HTA) was used to identify the gene expression changes in SF-Y3-treated EOC cells, which results were confirmed by quantitative real-time PCR (qPCR). Western blot and XBP1 RNA splicing PCR were performed to assess the activation of proteins in UPR pathway. Using 4u8c, an inhibitor of the ER transmembrane protein IRE1, we determined whether inhibiting UPR could rescue cancer cells from the SF-Y3-induced apoptosis. Co-immunoprecipitation (co-PI) was used to determine the effects of SF-Y3 on the ER membrane protein-protein interaction. Moreover, we encapsulated SF-Y3 with nanoparticle to improve its bioavailability for evaluating the in vivo efficacy in EOC mouse model as a single treatment and in combination with platinum-based chemotherapy. The cell viability data demonstrated that SF-Y3 significantly reduced the viability of EOC cells expressing high levels of Bip1, a key chaperone protein in the endoplasmic. SF-Y3 was less effective in EOC cells with low levels of Bip1 and has no effects on normal FT cells. P-S6 and Annexin V staining assays demonstrated that SF-Y3 inhibited EOC cell proliferation and induced apoptosis. HTA and qPCR data showed that the UPR genes were unregulated by SF-Y3. Western blot and XBP1 RNA splicing PCR results indicated that SF-Y3 activated proteins in the UPR pathway, including ATF6, PERK, eIF2α, XBP1, and CHOP. SF-Y3 interrupted the interaction between Bip1 and three ER membrane-associated sensors, supporting that Bip1 is a possible target of SF-Y3. UPR inhibitor 4u8c partially rescued the apoptosis induced by SF-Y3. These data support that SF-Y3 has anticancer activity in EOC models possibly through inhibiting Bip1 and inducing UPR-induced apoptosis. Further investigation of how SFs interact with Bip1 and UPR pathway in vitro and in vivo may lead to new approaches to overcome drug resistance and a significant therapeutic advance for EOC. Citation Format: Wonmin Park, Tobias MP. Hartwich, Kay Y. Chong, Chunming Liu, David S. Watt, Dongin Kim, Yang Yang-Hartwich. TARGETING UNFOLDED PROTEIN RESPONSE FOR OVARIAN CANCER THERAPY [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr NT-116.