Abstract：
Dysferlin (DYSF) has drawn much attention due to its involvement in dysferlinopathy and was reported to affect monocyte functions in recent studies. However, the role of DYSF in the pathogenesis of atherosclerotic cardiovascular diseases (ASCVD) and the regulation mechanism of DYSF expression have not been fully studied. In this study, Gene Expression Omnibus (GEO) database and epigenome-wide association study (EWAS) literatures were searched to find the DNA methylation-driven genes (including DYSF) of ASCVD. The hub genes related to DYSF were also identified through weighted correlation network analysis (WGCNA). Regulation of DYSF expression through its promoter methylation status was verified using peripheral blood leucocytes (PBLs) from ASCVD patients and normal controls, and experiments on THP1 cells and Apoe-/- mice. Similarly, the expressions of DYSF related hub genes, mainly contained SELL, STAT3 and TMX1, were also validated. DYSF functions were then evaluated by phagocytosis, transwell and adhesion assays in DYSF knock-down and overexpressed THP1 cells. The results showed that DYSF promoter hypermethylation up-regulated its expression in clinical samples, THP1 cells and Apoe-/- mice, confirming DYSF as a DNA methylation-driven gene. The combination of DYSF expression and methylation status in PBLs had a considerable prediction value for ASCVD. Besides, DYSF could enhance the phagocytosis, migration and adhesion ability of THP1 cells. Among DYSF related hub genes, SELL was proven to be the downstream target of DYSF by wet experiments. In conclusion, DYSF promoter hypermethylation upregulated its expression and promoted monocytes activation, which further participated in the pathogenesis of ASCVD.
摘要：
Dysferlin（DYSF）由于其参与了异常铁蛋白病而引起了广泛的关注，并且在最近的研究中报道了其影响单核细胞功能。然而,DYSF在动脉粥样硬化性心血管疾病(ASCVD)发病机制中的作用及DYSF表达的调控机制尚未得到充分研究。在这项研究中,通过基因表达综合(GEO)数据库和全表观基因组关联研究(EWAS)文献寻找ASCVD的DNA甲基化驱动基因(包括DYSF)。还通过加权相关网络分析(WGCNA)鉴定了与DYSF相关的hub基因。使用ASCVD患者和正常对照的外周血白细胞（PBLs）验证了通过其启动子甲基化状态对DYSF表达的调节，以及THP1细胞和Apoe-/-小鼠的实验。同样，DYSF相关hub基因的表达，主要包含销售，STAT3和TMX1也得到了验证。然后通过吞噬作用评估DYSF功能，DYSF敲低和过表达THP1细胞中的transwell和粘附测定。结果表明,DYSF启动子高甲基化上调其在临床样本中的表达,THP1细胞和Apo-/-小鼠，确认DYSF为DNA甲基化驱动基因。PBLs中DYSF表达和甲基化状态的结合对ASCVD具有相当的预测价值。此外,DYSF可以增强吞噬作用，THP1细胞的迁移和粘附能力。在DYSF相关的枢纽基因中，通过湿实验证明，销售是DYSF的下游目标。总之,DYSF启动子甲基化上调其表达并促进单核细胞活化,进一步参与了ASCVD的发病机制。
