Abstract 5740: Disruption of the acetylcholine signaling pathway inhibits angiogenesis and tumor growth in human lung adenocarcinoma

Abstract Purpose of the study: Lung adenocarcinoma (LUAD) accounts for about 60% of all lung cancers and shows a strong etiological correlation with smoking. Nicotine, the addictive component of tobacco smoke is known to accelerate the growth of human lung tumors by stimulating tumor angiogenesis. The pro-angiogenic effects of nicotine are mediated by nicotinic acetylcholine receptors (nAChRs). The endogenous ligand for nicotine is the neurotransmitter acetylcholine (ACh). The primary aim of this study was to determine if disruption of ACh production (via inhibition of the enzyme choline acetyltransferase; ChAT) could inhibit angiogenesis and suppress the growth of human LUADs. We used a small molecule inhibitor of ChAT namely BW813U for our studies. Experimental procedures. Primary human microvascular endothelial cells from the lung (HMEC-L) were used for all the experiments. These cells were used between passage 3-7. The anti-angiogenic activity of BW813U was measured in cell culture models by using the “MATRIGEL ASSAY”. The results obtained from the “MATRIGEL ASSAY” were confirmed using chicken chorioallantoic membrane (CAM) angiogenesis experiments. Finally, the anti-tumor activity of BW813U was measured in vivo by using xenograft model systems in SCID mice. Our studies also investigated the signaling pathways underlying the anti-angiogenic activity of BW813U, using chemical inhibitors and siRNA methodology. Results: BW813U robustly suppressed angiogenesis in Matrigel assays and CAM model systems. The administration of BW813U potently decreased the growth rate of H838 tumors xenotransplanted in SCID mice. Immunohistochemical staining experiments revealed that the anti-tumor activity of BW813U was correlated with decrease of CD31 angiogenic biomarker in H838 tumor sections. The anti-angiogenic activity of BW813U was mediated by the alpha7-nAChR pathway and involved the nitric oxide pathway. Taken together, our studies show that ChAT may be a valuable drug target for the therapy of LUAD. Conclusions: ChAT antagonists like BW813U decrease the growth of LUAD by suppressing tumor angiogenesis. Support or Funding Information: Funding for our study was supported by a NIH R15-AREA Grant (2R15CA161491-03) and a Center for Natural Products pilot grant from the WV-INBRE Grant Program (P20GM103434, PI:Gary Rankin). Furthermore, this study was supported in part by an Institutional Development Award (IDeA) Grant number P20GM104932 from the National Institute of General Medical Sciences (NIGMS) and the Research Core B of COBRE, a component of the National Institutes of Health (NIH). Citation Format: Piyali Dasgupta, Eric W. Bow, Krista L. Denning, Rama S. Gadepalli, John M. Rimoldi, Yi Charlie Chen, Sarah L. Miles. Disruption of the acetylcholine signaling pathway inhibits angiogenesis and tumor growth in human lung adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 5740.