Abstract 5327: Interleukin-1 upregulates dual oxidase 2 expression and ROS production in human pancreatic cancer cells

Abstract

Abstract Chronic pancreatitis increases the risk of developing pancreatic ductal adenocarcinoma (PDAC) and is associated with enhanced expression of the NADPH oxidase (NOX) isoform dual oxidase 2 (DUOX2). Interleukin-1 (IL-1) is a major upstream pro-inflammatory cytokine secreted by malignant or microenvironmental cells that supports pancreatic inflammation, tumorigenesis, invasiveness, and tumor heterogeneity. Previously, our laboratory demonstrated that the expression of DUOX2, one of seven NADPH oxidase (NOX) family members, is significantly enhanced by a wide variety of pro-inflammatory cytokines, and that DUOX2-related ROS production may contribute to the development of an oxidative, pro-angiogenic microenvironment in PDAC. In the current studies, we have shown that DUOX2 is upregulated and expressed along with its maturation factor, DUOXA2, in several human pancreatic cancer cell lines following exposure to both IL-1α and IL-1ß, two major agonists of IL-1 signaling. Administration of the anti-inflammatory antibody anakinra (an IL-1R1 antagonist) either in conjunction with or after prolonged treatment with IL-1 in vitro ablated induction of DUOX2/DUOXA2 and of IL-8 (CXCL8), a chemokine essential for neutrophil recruitment and a potent promoter of angiogenesis. Increased expression of other NOX family members following IL-1 exposure was not observed. Using CRISPR/Cas9-mediated DUOX2 knockout models, we report that DUOX2 is the major contributor to IL-1-mediated ROS production in PDAC cells. In a panel of PDAC lines, siRNA knockdown of proximal IL-1 signaling nodes, such as MYD88, significantly decreased expression of DUOX2/DUOXA2 and of CXCL8. However, knockdown of IRAK1 or IRAK4, two critical signaling kinases downstream of MYD88, as well as known downstream mediators of IL-1-related transcription (such as NF-κB, STAT1, MEK/ERK), did not affect IL-1-induced DUOX2/DUOXA2 upregulation, suggesting that alternative pathways of IL-1 signaling might be involved in the enhancement of DUOX2 expression by the cytokine. Because IL-1 signaling is known to affect various acetylation/methylation events in epithelial cells, we evaluated the effect of administering the histone acetyltransferase (HAT) inhibitor A-485 with IL-1; we found that A-485 significantly diminished cytokine-induced DUOX2/DUOXA2 upregulation; however, CXCL8 induction was unchanged. These studies suggest that epigenetic modulation of PDAC cells by IL-1 could contribute to DUOX2-induced ROS production and ROS-related inflammatory stress in the PDAC microenvironment. Citation Format: David J. Mallick, Yongzhong Wu, Mariam M. Konaté, Becky Diebold, Smitha Antony, Jennifer L. Meitzler, Guojian Jiang, Jiamo Lu, Krishnendu Roy, James H. Doroshow. Interleukin-1 upregulates dual oxidase 2 expression and ROS production in human pancreatic cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5327.