Pancreatic ductal adenocarcinoma (PDAC) is featured by the prevalence of large nerve bundles. Studies by us and others have demonstrated critical roles of the peripheral nervous system (PNS) in PDAC initiation, progression and metastasis. Using Trace-n-Seq, we transcriptionally profiled individual neurons innervating healthy pancreas and PDAC, revealing profound tumor-induced reprogramming of peripheral neurons. Functional nerve blockade significantly reduced tumor growth, demonstrating an active role of neural signaling in PDAC progression (Thiel et al., Nature 2025). Here, we investigate how PDAC cells respond to autonomic input and define the molecular circuits underlying cancer-nerve crosstalk, with the goal of identifying actionable targets to therapeutically disrupt neuro-tumoral interactions.

In vitro cultures of PDAC cells with peripheral ganglia explant was established and utilized for most experiments. The effects of neuronal presence on PDAC cell proliferation were examined using immunofluorescence microscopy. To determine whether direct contact or paracrine signaling plays a more significant role in neuron-cancer cell interaction, we utilize a range of methods including secretome analysis, super-resolution microscopy and electrophysiological recordings, as well as measurements of mRNA and protein expression.

The establishment of primary mouse autonomic ganglia cultures is technically demanding due to their small size, anatomical similarity to surrounding tissues, low abundance, and rapid overgrowth of non-neuronal cells in vitro. We successfully developed a robust isolation protocol yielding highly enriched ganglia preparations with minimal contamination. Using ganglia-PDAC co-culture systems, we demonstrated that sympathetic signaling directly enhances cancer cell proliferation. At sites of direct axon-tumor contact, interactions display neuronal hallmarks, including synapse-like structures and dynamic intracellular ion fluxes. Notably, PDAC cells in monoculture show comparable ion transients upon acute stimulation with autonomic neurotransmitters, indicating an intrinsic capacity to sense and respond to nerve-derived signals, independent of physical neuronal contact.

Our study identifies the autonomic nervous system as an active driver of PDAC progression and provides mechanistic evidence that PDAC cells are stimulated by neurons and can mount distinct, neuron-like functional responses to autonomic cues. We are currently delineating the molecular circuits underlying this bidirectional signaling and assessing their therapeutic tractability. Disrupting cancer-nerve communication may offer a novel and effective strategy to abrogate neural support of tumor growth and improve clinical outcomes in PDAC.

This project is supported by fundings from ERC (A. T., H. M.) and Dietmar Hopp Stiftung (HI-STEM gGmbH).