Abstract 6433: The interaction between netrin-1/neogenin modulates sympathetic innervation to promote tumorigenesis and accelerate pancreatic cancer progression

Abstract Introduction: Nerves are a major component of the tumor microenvironment with active crosstalk between cancer cells and facilitate cancer progression. Netrin-1 (Ntn1), an axon guidance molecule essential for neural development, has recently been implicated in the growth of several cancers, though its specific contribution to pancreatic cancer remains unclear. Methods: The expression of Ntn1 and its receptor Neogenin (Neo1) was investigated in spontaneous PDAC mouse models (LSL-Kras+/G12D/Pdx1-Cre; KC and LSL-Kras+/G12D/LSL-Trp53+/R172H/Pdx1-Cre; KPC). Additionally, the influence of Ntn1 on pancreatic cancer progression and innervation was evaluated using KC mice with or without conditional Ntn1 knockout (LSL-Kras+/G12D/Pdx1-Cre/Netrin1fl/fl; KCN). Murine PDAC cell lines and organoids were used to examine the functional roles of Ntn1 and Neo1 through genetic and pharmacological modulations. Metastatic capacity and effects on neurite outgrowth were studied in a liver metastasis model and in a co-culture system with Celiac Ganglions (CGs). Changes in innervation at primary and metastatic sites were analyzed using whole-mount staining with tissue clearance techniques. Results: Ntn1 and Neo1 expression was absent in normal acinar and ductal cells but was detected in PanIN lesions and PDAC, showing a positive correlation between them. PanIN progression rates were lower in KCN mice compared to KC mice, with reduced sympathetic innervation observed in KCN mice. In pancreatic organoids, KRAS activation increased Ntn1 and Neo1 expression, with recombinant Ntn1 (rNTN1) enhancing organoid formation and upregulating Sox2 and Sox9, suggesting a direct autocrine role in promoting stemness. Co-culture of CGs with Ntn1-overexpressing PDAC cells showed significant axonal elongation, supporting a role for Ntn1 in promoting innervation. Additionally, treatment with rNtn1 or Ntn1 overexpression facilitated epithelial-mesenchymal transition (EMT) in PDAC cells, an effect attenuated by Neo1 blockade. In a metastatic model, Ntn1 overexpression increased tumor burden and reduced survival, while Ntn1-neutralizing antibody or Neo1 knockdown improved outcomes. Nerve fiber density was significantly increased at PDAC liver metastasis sites with high Ntn1 expression. Conclusions: The Ntn1/Neo1 interaction is a key regulator of PDAC progression, directly influencing cancer cell stemness and EMT while indirectly modulating tumor innervation. Inhibiting Ntn1/Neo1 interactions reduced PDAC progression and improved survival, highlighting this interaction as a potential therapeutic target in pancreatic cancer. Citation Format: Yosuke Ochiai, Masaki Sunagawa, Hiroki Kobayashi, Feijing Wu, Jin Qian, Quin Waterbury, Biyun Zheng, Masahiro Hata, Yi Zeng, Hualong Zheng, Jonathan LaBella, Leah Zamechek, Timothy Wang. The interaction between netrin-1/neogenin modulates sympathetic innervation to promote tumorigenesis and accelerate pancreatic cancer progression [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 6433.