Abstract A08: Human preadipocytes promote pancreatic cancer cellular growth in a diabetes-specific manner

Abstract

Abstract Objective: Type 2 diabetes mellitus (DM) is a known risk factor for pancreatic ductal adenocarcinoma (PDAC), but the mechanisms underlying this association remain poorly understood. Multiple reports implicate adipocytes and preadipocytes as putative promoters of PDAC in humans and mice. To test whether preadipocytes regulate PDAC tumorigenic functions in the context of DM, we explored the effects of conditioned media (CM) prepared from preadipocytes from nondiabetic (NDM) and DM patients on PDAC cells. Methods: Preadipocytes were isolated from omental adipose tissue obtained from NDM and DM patients undergoing bariatric surgery. Serum-free conditioned media was prepared from these preadipocytes cultured in vitro for 72 hours. Sphere assays using T3M4 and Capan1 PDAC lines were performed in the presence or absence of conditioned media, and sphere surface area was measured over 2-3 weeks. Modifications of the sphere assays, including boiling of conditioned media as well as addition of neutralizing antibodies or recombinant proteins, were implemented. A proteome profiler array was used to identify differences in cytokine composition between CM from DM and NDM. A phospho-kinome array was used to identify differentially activated signaling pathways when PDAC cells were treated with NDM and DM conditioned media. Results: Conditioned media from human DM preadipocytes, compared to media from NDM preadipocytes, increased PDAC cell sphere surface area in both T3M4 and Capan1 cell lines. Sphere surface area correlated directly with hemoglobin A1c (HbA1c) but not sex or body mass index of the human subjects from which preadipocytes were derived. Boiling of preadipocyte supernatants abrogated their pro-growth effects. Proteome profiler array analysis identified different proteins that were differentially expressed by DM and NDM preadipocytes, among which chitinase-3-like protein 1 (CHI3L1) and interleukin-6 (IL-6) were increased over 2-fold in DM relative to NDM preadipocyte supernatants. Neutralizing antibody to human CHI3L1 and IL-6 attenuated the increase in PDAC sphere surface area in response to DM preadipocyte conditioned media in a dose-dependent manner. Recombinant human CHI3L1 and IL-6 enhanced the growth of PDAC cells when they were exposed to NDM conditioned media. Phospho-kinome analysis demonstrated that treatment of PDAC cells with DM-derived conditioned media increased activation of inflammation-associated signal transduction pathways. Conclusions: Preadipocyte-PDAC cell crosstalk regulates PDAC cell growth in a DM-specific and CHI3L1 and IL-6-dependent manner. These results suggest that the positive clinical correlation between DM and PDAC may be mediated by preadipocytes expressing higher levels of CHI3L1 and IL-6. Preadipocytes CHI3L1 and IL-6 represent targets for further research directed towards developing novel therapies for PDAC based on manipulation of the tumor microenvironment. Citation Format: Lawrence Delrosario, Carmen Flesher, Nicki Baker, Jenny Lazarus, Mirna Perusina-Lanfranca, Robert O'Rourke, Timothy Frankel. Human preadipocytes promote pancreatic cancer cellular growth in a diabetes-specific manner [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2019 Sept 6-9; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2019;79(24 Suppl):Abstract nr A08.