Abstract 6349A: Mechanical stress activates autophagy to induce drug resistance in pancreatic cancer cells

Abstract

Abstract INTRODUCTION: One of the most fundamental problems that oncologists are currently facing in pancreatic cancer patient care is the high resistance to all types of chemotherapies or targeted therapies. Thus, there is an urgent need for increasing patient response to treatment. A common characteristic among most pancreatic cancer patients that could serve as potential target for treatment, is the biomechanically abnormal tumor microenvironment (TME). This altered TME is characterized by an increased stiffness which among others, is responsible for the development of mechanical stresses in the tumor interior. Even though several studies have investigated the effect of mechanical stress on cancer cells and tumor progression, it has not yet defined whether it can affect drug sensitivity and what are the mechanisms involved. HYPOTHESIS: Based on our recent findings, obtained by a Reverse Phase Protein Assay, we observed that mechanical stress activates a resistance response, employing autophagy activation, in pancreatic cancer cells. To this end, we hypothesize that mechanical stress developed during tumor growth, can induce autophagy activation to eventually promote drug resistance in pancreatic tumors. Therefore, inhibition of this mechanism can improve therapy. METHODOLOGY: We use two pancreatic cancer cells lines, MIA PaCa-2 and PANC-1, to apply mechanical stress, either in a 2D experimental setup or by employing tumor spheroids in a 3D agarose matrix. In both cases, cells are exposed to the most common chemotherapeutic agents used for pancreatic cancer treatment, gemcitabine and 5-Fluorouracil (5FU). The cytotoxic effect of each drug, as well as autophagy- and signaling pathway- activation are then analyzed. Finally, in vitro assays and treatments, along with in vivo animal models of pancreatic tumors, are performed to test the efficacy of an autophagy inhibitor combined with chemotherapy and a mechanotherapeutic that modulates intratumoral mechanical stresses. RESULTS: Proteomic analysis, together with pharmacological inhibition and siRNA treatments, revealed that mechanical stress activates autophagy to impair cell apoptosis and promote cancer cell survival. This stress-adaptation mechanism in turn impairs the cytotoxic effect of both gemcitabine and 5FU. A combined treatment employing chemotherapy with an autophagy inhibitor, and a mechanotherapeutic agent, showed promising results in increasing the overall-survival of pancreatic tumor-bearing mice. CONCLUSION: Our results provide solid evidence that alleviation of intratumoral mechanical stresses with mechanotherapeutics combined with autophagy inhibitors can improve the efficacy of chemotherapeutic agents in pancreatic cancer. Citation Format: Maria Kalli, Ruxuan Li, Ioannis Zervantonakis, Triantafyllos Stylianopoulos. Mechanical stress activates autophagy to induce drug resistance in pancreatic cancer cells [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 6349A.