Abstract 3677: Early detection of pancreatic cancer using methylated circulating cell-free DNA of ZFP30 and ZNF781genes

Abstract Background: Pancreatic cancer (PC) is often diagnosed at advanced stages, leading to limited treatment options and poor survival rates. Current biomarkers, such as CA19-9, have high false-positive rates, with only 4 out of 1, 000 abnormal cases confirmed as cancer. Aberrant DNA methylation, a stable and early epigenetic event in tumorigenesis, has emerged as a promising biomarker for early detection and disease monitoring. Objective: This study presents a novel blood test analyzing methylated cell-free DNA (mcfDNA) of ZFP30 and ZNF781genes for the early detection of pancreatic cancer. Functional assays were also performed to explore the tumor-suppressive roles of ZFP30 and ZNF781 in pancreatic cancer cell lines. Methods: ZFP30 and ZNF781 were identified through analyses of plasma samples from Taiwanese pancreatic cancer patients combined with data from The Cancer Genome Atlas (TCGA). A novel test was developed to assess methylation levels in cfDNA extracted from plasma. Clinical validation was conducted on PC patients and healthy controls, while functional studies assessed the effects of exogenous ZFP30 and ZNF781 expression on cancer cell proliferation and migration. Results: The methylation levels of ZFP30 and ZNF781 were significantly elevated in 87% of pancreatic cancer (PC) tumor tissues compared to adjacent normal tissues in PC patients. Furthermore, these methylation markers were detected in circulating cell-free DNA (cfDNA) in 99% of plasma samples from PC patients, demonstrating an area under the curve (AUC) of 0.96, with a sensitivity of 99% and a specificity of 93.9%. Methylation signals were nearly undetectable in plasma from healthy subjects, and the test accurately identified PC patients across all stages. Comparative analysis demonstrated superior performance over traditional biomarkers such as CA19-9 and CEA. Functional studies showed that exogenous expression of ZFP30 and ZNF781significantly inhibited pancreatic cancer cell proliferation and migration, supporting their roles as tumor suppressor-like genes. Conclusion: The methylated cfDNA of ZFP30 and ZNF781 offers a highly sensitive and specific biomarker for the early detection of pancreatic cancer. Integrating this blood test into clinical practice could greatly enhance diagnostic accuracy. Additionally, the tumor suppressor-like functions of ZFP30 and ZNF781 highlight their potential as therapeutic targets, providing further avenues for pancreatic cancer management and treatment. Citation Format: Ruo-Kai Lin, Yao Yu Hsieh, Hsieh-Tsung Shen. Early detection of pancreatic cancer using methylated circulating cell-free DNA of ZFP30 and ZNF781genes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 3677.