Tumor cell differentiation is a critical determinant of malignancy and clinical treatment selection. Pancreatic ductal adenocarcinoma (PDAC), a poorly differentiated and highly aggressive tumor, has a poor prognosis, whereas well-differentiated tumors often correlate with better outcomes. The mechanisms underlying differentiation and its therapeutic potential remain unclear.

This study aims to investigate whether inducing transdifferentiation in pancreatic cancer cells can reduce malignancy, focusing on the role of the transcription factor NeuroD1 and its regulatory pathways.

We analyzed single-cell RNA-seq data from the GEO database to identify differentiation-associated genes. NeuroD1 was overexpressed in PDAC cells to assess its effects on transdifferentiation and proliferation. Drug screening and molecular docking were performed to identify differentiation-inducing compounds. RNA sequencing, coimmunoprecipitation, and mass spectrometry were used to identify NeuroD1-interacting proteins. Cell/patient-derived xenograft mouse models are utilized for in vivo experiments and compound efficacy testing.

Highly differentiated tumor cells exhibited elevated NeuroD1 expression. NeuroD1 overexpression promoted neuronal transdifferentiation and suppressed proliferation. Neuropathiazol, a neurogenic inducer, was found to bind MET and upregulate NeuroD1 via the PI3K/Akt pathway, enhancing transdifferentiation and inhibiting tumor growth. Neurog3 was identified as a functional partner of NeuroD1.

Our findings demonstrate that pancreatic cancer cells can be induced to transdifferentiate through NeuroD1 activation or pharmacological induction, suggesting a potential therapeutic strategy to mitigate malignancy by reprogramming tumor cells into less aggressive states.