摘要目的:基于DNA甲基化芯片与转录组测序(RNA-seq)数据，分析 KMT2A基因变异导致Wiedemann-Steiner综合征(WDSTS)患儿在表观遗传与基因表达层面的改变，筛选关键生物学通路及核心基因，以阐明其发病机制，并为临床诊疗提供理论依据。 方法:选取2016年11月至2024年12月于上海交通大学医学院附属上海儿童医学中心就诊的16例WDSTS患儿及10例健康对照儿童为研究对象，作回顾性分析。采集所有受试者外周血样本，提取基因组DNA和总RNA。采用全基因组DNA甲基化芯片检测甲基化水平，识别差异甲基化位点及其对应基因，并进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)通路富集分析；通过RNA-seq筛选差异表达基因，并开展GO与KEGG功能注释分析；整合甲基化与表达数据，对重叠差异关联基因进行GO、KEGG及基因-通路网络分析。本研究方案经上海交通大学医学院附属国际和平妇幼保健院医学伦理委员会批准(批准号：GKLW-A-2024-006-01)。结果:共检测出2 652个差异甲基化位点，对应1 262个基因，其中高甲基化基因833个(66%)，主要富集于细胞连接相关功能；低甲基化基因429个(34%)，显著参与神经系统发育及形态发生过程。转录组分析鉴定出2 627个差异表达基因，包括765个上调基因(29%)，主要涉及细胞免疫过程；1 862个下调基因(71%)，主要参与物质运输功能。整合分析发现93个基因在甲基化与表达层面均存在显著差异，这些基因富集于细胞外基质、钙离子结合、神经发育及细胞黏附等过程，进一步筛选出 LAMB1、 LAMB2、 NID1等关键基因，可能参与WDSTS的发病机制。 结论:整合DNA甲基化与转录组数据可揭示WDSTS特异的表观遗传-转录调控模式，为该病的机制解析与诊断策略优化提供新的理论基础。

abstractsObjective:To investigate epigenetic and transcriptional alterations in children with Wiedemann-Steiner syndrome (WDSTS) due to variants of KMT2A gene using genome-wide DNA methylation array and RNA sequencing (RNA-seq), and identify the key pathways and candidate genes. Methods:A retrospective study was carried out for 16 children with WDSTS and 10 healthy controls who visited Shanghai Children′s Medical Center, Shanghai Jiao Tong University School of Medicine between November 2016 and December 2024. Peripheral blood samples were collected. Genomic DNA and total RNA were extracted using commercially made kits. Genome-wide DNA methylation profiling was conducted to identify differentially methylated positions (DMPs) and annotated genes, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. RNA-seq was performed to identify differentially expressed genes (DEGs) and conduct GO/KEGG functional annotation. Methylation and expression data were integrated to identify overlapping genes showing significant changes at both levels, followed by GO, KEGG and gene-pathway network analyses. This study was approved by the Ethics Committee of the hospital (Ethics No.: GKLW-A-2024-006-01).Results:A total of 2 652 DMPs corresponding to 1 262 genes were identified, which included 833 hypermethylated genes (66%) and 429 hypomethylated genes (34%). Hypermethylated genes were mainly enriched for functions related to cell junctions, while hypomethylated genes were significantly involved in nervous system development and morphogenesis. RNA-seq identified 2 627 DEGs, including 765 up-regulated genes (29%) and 1 862 down-regulated genes (71%). Up-regulated genes were mainly associated with immune-related processes, and down-regulated genes were mainly related to substance transport. Integrative analysis identified 93 overlapping genes with significant changes in both methylation and expression. And these genes were enriched in extracellular matrix-related processes, calcium ion binding, neurodevelopment, and cell adhesion. Key candidate genes, including LAMB1, LAMB2 and NID1, were further prioritized. Conclusion:Integrated analysis of DNA methylation and transcriptome data reveals WDSTS-related epigenetic-transcriptional alterations and provides clues for exploring disease mechanisms and optimizing diagnostic strategies.