Abstract 3667: Attenuation of NF-κB signaling in myeloid cells enhances urethane-induced lung tumorigenesis via neutrophil-derived IL-1β

Abstract

Abstract The transcription factor Nuclear Factor κB (NF-κB) is a master regulator of inflammation, and its activation is associated with tumorigenesis in a variety of cancers. Our group and others have shown that NF-κB signaling in lung epithelial cells is critical for lung carcinogenesis. However, pharmaceutical inhibition of NF-κB signaling has not been effective for the treatment of lung tumorigenesis in animals or humans for reasons that remain unknown. We hypothesized that attenuation of NF-κB signaling in myeloid cells contributes to inefficacy of NF-κB-targeted therapies by unmasking pro-tumorigenic characteristics of inflammatory cells. For our studies, we utilized mice with IKKβ deleted specifically in myeloid cells (IKKβΔmye) and induced lung tumorigenesis with a single intraperitoneal injection of the lung carcinogen urethane (1g/kg). We found that IKKβΔmye mice developed more atypical adenomatous hyperplasia (AAH) lesions at 6 weeks and more lung tumors at 16 weeks after urethane compared to WT mice. These histological differences were correlated with enhanced lung inflammation in IKKβΔmye mice which was observed 1 week after urethane and persisted for up to 6 weeks. Flow cytometric analysis of myeloid cell populations revealed that IKKβΔmye mice contained more pulmonary neutrophils at 1 and 6 weeks after urethane compared to WT mice. In bone marrow chimeras generated by transplantation of bone marrow from IKKβΔmye or WT donors into recipient NF-κB reporter mice, we showed that depletion of neutrophils using Ly6G antibody treatment (100 ug/twice weekly IP injection) during the first 6 weeks of lung carcinogenesis blocked NF-κB activation in stromal cells and reduced tumor multiplicity in IKKβΔmye mice to levels observed in urethane-treated WT mice. Thus, these studies indicated that neutrophils play an important role in lung tumorigenesis during early tumor development and may do so by limiting NF-κB activation in lung epithelial cells. Evaluation of cytokines in the lungs of WT and IKKβΔmye mice at 1 week after urethane revealed higher levels of IL-1β in IKKβΔmye mice compared to WT mice. Further examination of myeloid cells sorted 1 week after urethane identified neutrophils as the key producers of IL-1β in the lungs of IKKβΔmye mice. Neutralization of IL-1 signaling by IL-1 receptor antagonist (IL-1ra) treatment during the first 4 weeks of carcinogenesis decreased the number of AAH lesions in IKKβΔmye mice 6 weeks after urethane and reduced the number of tumors at 16 weeks in IKKβΔmye mice. Taken together, our data suggest that neutrophils can support tumor promotion through production and secretion of IL-1β, which may activate pro-tumorgenic NF-κB signaling in the lung epithelium. We speculate that the paradoxical increase in inflammation and IL-1β production resulting from NF-κB inhibition in myeloid cells contributes to the lack of effectiveness of NF-κB inhibitors in patients with lung cancer. Citation Format: Allyson McLoed, Rinat Zaynagetdinov, Taylor Sherrill, Fiona Yull, Timothy Blackwell. Attenuation of NF-κB signaling in myeloid cells enhances urethane-induced lung tumorigenesis via neutrophil-derived IL-1β. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3667. doi:10.1158/1538-7445.AM2014-3667