Skeletal muscle tissue RNA-Seq and RRBS in T2D, Obese and Active subjects

We performed RNA-Seq on skeletal muscle tissue in Active individuals, indivduals with obesity (Obese) and individuals with obesity and T2D (T2D) (n =6 per group). PCA of top ranked contributing genes show clear separation of the Active group from Obese and T2D with considerable overlap between Obese and T2D. Over-representation analysis highlighted that Active had an upregulation of genes related to respiration/mitochondrial capacity in comparison to Obese and T2D. RRBS was performed on skeletal muscle tissue from a subset of participants from each group (n = 3). Data was analyzed to reveal (DMS) differentially methylated sites located at differentially expressed genes related to mitochondrial respiration/function. Out of a combined 488 DMS related to mitochondrial related differentially expressed genes, 11% were significantly positively correlated with gene expression and 12% were significantly negatively associated with gene expression. Significantly correlated DMS were mainly located in the promoter and 1st intron region Skeletal muscle tissue from M. Vastus Lateralis was obtained from lean Active individuals, individuals with Obesity and individuals with T2D and Obesity to perfrom transcriptomics by RNA-Seq and DNA methylation by RRBS.

本研究对瘦体型健康活跃个体、肥胖个体以及肥胖合并2型糖尿病（Type 2 Diabetes, T2D）个体的骨骼肌组织开展RNA测序（RNA-Seq），每组纳入6名受试者（n=6）。对贡献度排名靠前的基因进行主成分分析（Principal Component Analysis, PCA）结果显示，瘦体型健康活跃组与肥胖组、T2D组呈现明显分离，而后两组间存在较多重叠。富集分析（Over-representation Analysis）结果表明，相较于肥胖组与T2D组，瘦体型健康活跃组的呼吸/线粒体功能相关基因呈现上调表达。本研究同时对每组的部分受试者（n=3）的骨骼肌组织开展简化代表性亚硫酸氢盐测序（Reduced Representation Bisulfite Sequencing, RRBS）。数据分析显示，差异甲基化位点（Differentially Methylated Sites, DMS）富集于线粒体呼吸/功能相关的差异表达基因区域。在共计488个与线粒体相关差异表达基因相关的DMS中，11%与基因表达呈显著正相关，12%与基因表达呈显著负相关。这类显著相关的DMS主要分布于启动子区域及第一内含子区。本研究的骨骼肌样本均取自股外侧肌（Musculus Vastus Lateralis, M. Vastus Lateralis），受试者包括瘦体型健康活跃个体、肥胖个体以及肥胖合并T2D个体，通过RNA-Seq开展转录组学分析、通过RRBS开展DNA甲基化检测。