Zeybel Müjdat, Hardy Timothy, Robinson Stuart M, Fox Christopher, Anstee Quentin M, Ness Thomas, Masson Steven, Mathers John C, French Jeremy, White Steve, Mann Jelena
Institute of Cellular Medicine, Faculty of Medical Sciences, Newcastle University, 4th Floor William Leech Building, Framlington Place, Newcastle upon Tyne, NE2 4HH UK.
Cellular Pathology Department, Royal Victoria Infirmary, Newcastle upon Tyne Hospitals NHS Foundation Trust, Queen Victoria Road, Newcastle upon Tyne, NE1 4LP UK.
Clin Epigenetics. 2015 Mar 14;7(1):25. doi: 10.1186/s13148-015-0056-6. eCollection 2015.
Chronic liver injury can lead to the development of liver fibrosis and cirrhosis but only in a minority of patients. Currently, it is not clear which factors determine progression to fibrosis. We investigated whether DNA\methylation profile as determined by pyrosequencing can distinguish patients with mild from those with advanced/severe fibrosis in non-alcoholic liver disease (NAFLD) and alcoholic liver disease (ALD). To this end, paraffin-embedded liver biopsies were collected from patients with biopsy-proven NAFLD or ALD, as well as paraffin-embedded normal liver resections, genomic DNA isolated, bisulfite converted and pyrosequencing assays used to quantify DNA methylation at specific CpGs within PPARα, PPARα, TGFβ1, Collagen 1A1 and PDGFα genes. Furthermore, we assessed the impact of age, gender and anatomical location within the liver on patterns of DNA methylation in the same panel of genes.
DNA methylation at specific CpGs within genes known to affect fibrogenesis distinguishes between patients with mild from those with severe fibrosis in both NAFLD and ALD, although same CpGs are not equally represented in both etiologies. In normal liver, age, gender or anatomical location had no significant impact on DNA methylation patterns in the liver.
DNA methylation status at specific CpGs may be useful as part of a wider set of patient data for predicting progression to liver fibrosis.
慢性肝损伤可导致肝纤维化和肝硬化的发生，但仅在少数患者中出现。目前，尚不清楚哪些因素决定了纤维化的进展。我们研究了焦磷酸测序测定的DNA甲基化谱是否能够区分非酒精性肝病（NAFLD）和酒精性肝病（ALD）中轻度纤维化患者与重度/晚期纤维化患者。为此，我们收集了经活检证实的NAFLD或ALD患者的石蜡包埋肝活检组织，以及石蜡包埋的正常肝切除组织，分离基因组DNA，进行亚硫酸氢盐转化，并使用焦磷酸测序测定法对PPARα、PPARα、TGFβ1、胶原蛋白1A1和PDGFα基因内特定CpG位点的DNA甲基化进行定量。此外，我们评估了年龄、性别和肝脏内解剖位置对同一组基因DNA甲基化模式的影响。
已知影响纤维化形成的基因内特定CpG位点的DNA甲基化能够区分NAFLD和ALD中轻度纤维化患者与重度纤维化患者，尽管相同的CpG位点在两种病因中并不具有同等代表性。在正常肝脏中，年龄、性别或解剖位置对肝脏中的DNA甲基化模式没有显著影响。
特定CpG位点的DNA甲基化状态作为更广泛的患者数据的一部分，可能有助于预测肝纤维化的进展。
