Sensory Neuron Rewiring in Pancreatic Ductal Adenocarcinoma: from Single-Cell RNA Profiling to Extracellular Vesicles-based strategies

Background & Aims: Neuronal innervation of the pancreas has historically been characterized using marker-based classification and physiological studies, but its transcriptomic landscape remains only partially explored. A detailed molecular profile of pancreatic sensory neurons could provide insights into their role in health and disease, particularly in pancreatic ductal adenocarcinoma (PDAC), where neural remodeling influences tumor progression and pain signaling. Methods: Wild-type and PDAC mice were injected with the retrotracer Fast Blue into pancreatic or cancerous tissue. Dorsal root ganglia were cultured, and Fast Blue-positive sensory neurons were isolated, lysed, and analyzed using single-cell RNA sequencing. Data was validated using immunohistochemistry, organoid cultures and qPCR. Results: We performed transcriptomic profiling of sensory neurons innervating the pancreatic head and tail under normal and cancer conditions. Our analysis identified neurofilament-containing neurons (NF) as the predominant sensory subtype in both contexts, while non-peptidergic neurons (NP) were underrepresented in tumor-associated innervation. Differential gene expression analysis revealed a unique subset of genes upregulated in sensory neurons innervating pancreatic tumors, many linked to mitochondrial activity. Further validation also revealed the presence of transcripts transferred via extracellular vesicles (including a non-coding portion of the PDX1 gene), suggesting a novel mechanism of tumor-neuron interaction. Conclusions: Our findings provide a detailed characterization of pancreatic and pancreatic ductal adenocarcinoma sensory innervation. We identified a circulating long non-coding RNA of potential tumor origin in the PDAC mouse model, revealing new therapeutic and biomarker opportunities in pancreatic cancer.