Abstract 3427: Targeting MUC13 to overcome the survival mechanisms for improved response to chemotherapy
Authors: Saini Setua, Sheema Khan, Murali M. Yallapu, Stephen W. Behrman, Nadeem Zafar, Meena Jaggi, Subhash C. Chauhan
Journal: Cancer Research (2017)

Abstract Background: Pancreatic cancer (PanCa) is a third leading cause of cancer-related deaths in the US due to late diagnosis and development of chemoresistance. Therefore, understanding molecular mechanisms that confer survival benefit to PanCa cells may offer new therapeutic strategies for PanCa treatment.Mucin, MUC13 aberrantly expressed in PanCa,promoting cancer growth and progression and these effects are abrogated by microRNA-145 (miR-145) restoration. Unlike other cancer types, PanCa is highly resistant to Tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL) that emerges as one of the most promising experimental cancer therapeutic drugs.Herein, we demonstrate the integration of novel approach to overcome chemoresistance and offer TRAIL-based therapeutic strategies. Methods: MUC13 expressing and null stable PanCa cells were generated to investigate the role of MUC13 in cell survival.miR-145 mimics were used to investigate the effect of MUC13 silencing in promoting survival and inhibiting apoptosis in presence of TRAIL using Western blotting, cell proliferation (MTT), Acridine orange staining and flow cytometry apoptosis assays(cell cycle, Annexin V/7AAD staining). Various distinct domain specific constructs of MUC13 were constructed such as the mucin (α), Beta (β) sea urchin sperm protein enterokinase arginine (SEA) domain and cytoplasmic (CD) domains and transfected into MUC13 null Panc-1 cells to identify the role of different domains in eliciting survival benefit to PanCa cells. Results: Results demonstrate that MUC13 expression blocks activation of caspase-8 and death receptor-mediated apoptosis in PanCa cells in response toTRAIL treatment as observed through Western blotting and flow cytometer. Inhibition of MUC13 using shRNA knockdown or miR-145 restoration resulted in the TRAIL-mediated increase in apoptotic cell death as evidenced by AnnexinV/ 7AAD and sub G0 population, as well as rendered PanCa cells sensitive to treatment with drugs, such as paclitaxel. Additionally, cells treated with TRAIL in combination with paclitaxel or Abraxane showed enhanced apoptosis on inhibition of MUC13 expression using miR-145 restoration. Further investigation showed that cytoplasmic domain of MUC13 (MUC13-CD) is indispensable for blockingcaspase-8 activation and PARP cleavage, indicating that the MUC13-CD blocks TRAIL-induced signaling upstream to Bid by inhibiting caspase-8 activation. Conclusion: These observations suggest that MUC13 contributes to the survival advantage in PanCa cells in response to treatment with drugs or death inducing ligands such as, Tumor necrosis factor related apoptosis-inducing ligand (TRAIL) which can be strategically overcome by miR-145 replenishment. These findings indicate that MUC13 silencing sensitizes PanCa cells towards TRAIL therapy and counteracts chemoresistance mechanisms in PanCa that may lead to novel combination therapies for PanCa treatment. Citation Format: Saini Setua, Sheema Khan, Murali M. Yallapu, Stephen W. Behrman, Nadeem Zafar, Meena Jaggi, Subhash C. Chauhan. Targeting MUC13 to overcome the survival mechanisms for improved response to chemotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3427. doi:10.1158/1538-7445.AM2017-3427