Sympathetic nerve–fibroblast crosstalk drives nerve injury, fibroblast activation, and matrix remodeling in pancreatic cancer

Pancreatic cancer is a highly innervated gastrointestinal disease in which sympathetic nerves play a critical role in modulating tumor growth and the tumor microenvironment (TME). While recent studies suggest that sympathetic nerves influence various TME components, including lymphoid and myeloid immune cells, their interactions with cancer-associated fibroblasts (CAFs) remain poorly understood. CAFs are a hallmark of pancreatic tumors and are known to upregulate axon guidance and neuroactive cues, suggesting a potential feedback loop with tumor-innervating nerves. Here, we investigated the bidirectional crosstalk between sympathetic nerves and CAFs in human and mouse pancreatic tumors. Using a chemo-genetic ablation model, we selectively eliminated pancreatic sympathetic nerves and found that denervation significantly reduced tumor size in female mice. To further dissect this interaction, we established co-culture systems with immortalized pancreatic fibroblasts and primary sympathetic neuron explants, identifying key transcriptional changes driven by CAF-sympathetic nerve signaling. Our findings demonstrate that sympathetic signaling enhances CAF activation and extracellular matrix remodeling, while activated CAFs, in turn, induce transcriptional programs in sympathetic neurons associated with nerve injury response. These results establish CAFs as central mediators of the tumor-supportive role of sympathetic nerves, offering new insights into the neural regulation of pancreatic cancer progression. Overall design: In this experiment, we indirectly cocultured p1-p4 murine pup superior cervical ganglia (SCG) with pancreatic fibroblasts, immortalized pancreatic stellate cells (PSCs), and control fibroblasts, mouse embryonic fibroblasts (MEFs, [NIH/3T3 CRL-1658]). Between 8 and 13 explants were plated in a coated 6-well plate and allowed to grow with 10.0 X 103 PSC or MEF cell lines on cell culture inserts (Falcon; 353090) for about 70 hours. RNA was isolated using the RNeasy Kit (Qiagen; 74104) for RNA purification. The samples: Nerve (SCG alone), Nerve_PSC (SCG cocultured with PSCs), Nerve_MEF (SCG cocultured with MEFs), PSC (PSCs alone), and PSC+N (PSCs cocultured with SCGs).