Abstract 6367: Activation of vitamin D/VDR signaling reverses gemcitabine resistance of pancreatic cancer cells through inhibition of MUC1 expression

Abstract

Abstract Objective: Pancreatic ductal adenocarcinoma (PDA) has a dismal prognosis, largely due to its resistance to currently available therapies. Identifying the determinants of and signaling pathways regulating this resistance is essential for the development of effective therapeutic modalities to improve the outcome of PDA. The purpose of this study is to define the role of vitamin D/VDR signaling and underlying mechanisms in PDA pathogenesis and therapeutic resistance. Materials & Methods: We used an unbiased reverse phase protein array (RPPA) to identify Vitamin D3 downstream molecules; we also used a variety of cellular and molecular biological approaches to determine the impact of vitamin D/VDR signaling on the expression of MUC1 mucin and the cellular response to gemcitabine treatment in PDA cells. We used in vivo experimental systems to determine the effect of combination use of gemcitabine with Vitamin D analogue on pancreatic tumorigenicity. Additionally, we used ultra-high resolution mass spectrometry (HRMS) to analyze the relative abundance of intracellular dFdCTP in tumor tissue samples. Results: We identified that MUC1 protein expression was significantly reduced in AsPC-1 human pancreatic cancer cells after treatment with vitamin D3 or its analogue calcipotriol (Cal) by the RPPA analysis, which was further confirmed in MiaPaca-2 and Colo357 pancreatic cancer cells. VDR negatively regulated MUC1 expression in pancreatic cancer cells in both gain- and loss-of-function assays. We established gemcitabine (Gem) resistant AsPC-1-GemR and Colo357-GemR cells and found that AsPC-1-GemR and Colo357-GemR cells had much lower levels of VDR but higher levels of MUC1 protein expression compared to their parental cells, Vitamin D3 or its analogue significantly induced VDR and inhibited MUC1 expression in AsPC-1-GemR and Colo357-GemR cells, and sensitized the resistant cells to gemcitabine treatment. The results of siRNA knockdown experiments indicated that inhibition of MUC1 expression was essential for Cal’s sensitization of PDA cells to Gem treatment in vitro. In keeping with these findings, administration of the Vitamin D3 analogue, paricalcitol (PA) significantly enhanced the therapeutic efficacy of Gem compared to chemotherapy alone in xenograft and syngeneic mouse models of pancreatic cancer without any significant systemic toxicity. The improved combination treatment correlated with increased intratumoral concentration of dFdCTP, the biological active metabolite of gemcitabine. Conclusions: This study reveals a previously unidentified vitamin D/VDR-MUC1 signaling pathway involved in the regulation of gemcitabine resistance of PDA. Our results suggest that combinational therapies which include targeted activation of vitamin D/VDR signaling may improve the outcomes of patients with PDA. Citation Format: Daoyan Wei, Margarete Hafley, Liang Wang, Yi Liu, Peiying Yang, Xiangsheng Zuo, Robert S. Bresalier. Activation of vitamin D/VDR signaling reverses gemcitabine resistance of pancreatic cancer cells through inhibition of MUC1 expression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6367.