Integrative identification and functional interrogation of DNA methylation-mediated epigenetic gene alterations in pancreatic cancer

Background: Pancreatic ductal adenocarcinoma (PDAC) remains a highly lethal malignancy with limited treatment options and a lack of effective molecular targets. Recent studies have highlighted the critical role of epigenetic modifications in the pathogenesis of various cancers. Methods: To identify targetable epigenetic modifications in PDAC, we analyzed publicly available genomic datasets from pancreatic cancer patients and conducted genomic analysis of pancreatic cancer cell lines and patient-derived cell lines. The functional significance of the identified epigenetic modification was assessed by modulating its expression through DNA demethylation and overexpression experiments. Results: We found that neurofilament (NEFL) was consistently and significantly downregulated in both pancreatic cancer patients and cell lines. We restored the expression of NEFL by DNA demethylation and overexpression in pancreatic cancer cell lines and found that NEFL overexpression reduced cancer cell proliferation, colony formation, invasion, and migration. Finally, we performed RNA sequencing to analyze the changes in gene expression profiles following NEFL overexpression. Discussion: Our findings indicate that the epigenetic silencing of NEFL plays a crucial role in pancreatic caner progression. This study highlights the potential of targeted modification of DNA methylation as a promising novel therapeutic strategy for PDAC. Key words: Pancreatic cancer; Neurofilament; DNA hypermethylation; epigenetic modification.