解读胰腺导管腺癌的异质性和随后的治疗选择仍然是一个挑战。我们的目的是表征胰腺导管腺癌进展中涉及的表观遗传调控途径。对胰腺癌患者组织和细胞系进行整体 DNA 甲基化分析，以确定差异甲基化基因。采用靶向亚硫酸氢盐测序和体外甲基化报告基因测定来研究位点特异性甲基化和转录调控之间的直接联系。使用一系列体外功能丧失和功能获得研究以及体内异种移植和 KPC（LSL-Kras G12D/；LSL-Trp53 R172H/；Pdx1-Cre）小鼠模型来评估胰腺癌细胞特性。在 3 个不同的胰腺癌患者队列中进行基因和蛋白表达分析，并与临床病理参数相关。我们将肝细胞核因子 4A (HNF4A) 确定为胰腺癌高甲基化的新靶标，并证明位点特异性近端启动子甲基化驱动 HNF4A转录抑制。患者的表达分析表明胰腺癌组织中 HNF4A 表达的甲基化相关抑制。体外和体内研究表明，HNF4A 是胰腺癌中的一种新型肿瘤抑制因子，可调节癌症的生长和侵袭性。 KPC 小鼠模型和人类胰腺癌组织中都证明，HNF4A 表达在疾病早期显着下降。最重要的是，HNF4 缺失与患者总体生存率较差相关。由启动子 DNA 甲基化介导的 HNF4A 沉默会促进胰腺癌的发展和侵袭性，导致患者生存率较差。© 2024 作者。

Decoding pancreatic ductal adenocarcinoma heterogeneity and the consequent therapeutic selection remains a challenge. We aimed to characterize epigenetically regulated pathways involved in pancreatic ductal adenocarcinoma progression.Global DNA methylation analysis in pancreatic cancer patient tissues and cell lines was performed to identify differentially methylated genes. Targeted bisulfite sequencing and in vitro methylation reporter assays were employed to investigate the direct link between site-specific methylation and transcriptional regulation. A series of in vitro loss-of-function and gain-of function studies and in vivo xenograft and the KPC (LSL-Kras G12D/+ ; LSL-Trp53 R172H/+ ; Pdx1-Cre) mouse models were used to assess pancreatic cancer cell properties. Gene and protein expression analyses were performed in 3 different cohorts of pancreatic cancer patients and correlated to clinicopathological parameters.We identify Hepatocyte Nuclear Factor 4A (HNF4A) as a novel target of hypermethylation in pancreatic cancer and demonstrate that site-specific proximal promoter methylation drives HNF4A transcriptional repression. Expression analyses in patients indicate the methylation-associated suppression of HNF4A expression in pancreatic cancer tissues. In vitro and in vivo studies reveal that HNF4A is a novel tumor suppressor in pancreatic cancer, regulating cancer growth and aggressiveness. As evidenced in both the KPC mouse model and human pancreatic cancer tissues, HNF4A expression declines significantly in the early stages of the disease. Most importantly, HNF4 loss correlates with poor overall patient survival.HNF4A silencing, mediated by promoter DNA methylation, drives pancreatic cancer development and aggressiveness leading to poor patient survival.© 2024 The Authors.