Genomics and Child Health, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK.
Populations, Evidence, and Technologies, Division of Health Sciences, Warwick Medical School, University of Warwick, Coventry, UK.
Epigenetics. 2022 Oct;17(10):1219-1233. doi: 10.1080/15592294.2021.2003043. Epub 2021 Nov 25.
Vitamin B12 has multiple biochemical functions including in the one-carbon cycle generating a methyl group for DNA methylation, and metabolism of fatty acids and amino acids to generate energy via the citric acid cycle. The aim of our study was to use a combined epigenomic and transcriptomic approach to identify novel genes mediating the effect of B12 on adipogenesis.Human pre-adipocytes (CHUB-S7) were treated with a range of B12 (0-500 nM) concentrations from the day of cell seeding until harvesting in discovery and validation experiments prior to genome-wide methylation analysis using the Illumina HumanMethylation 450Beadchip. For transcriptomic analysis, RNA-seq libraries were run on the Illumina HiSeq 2500. To further investigate the expression of any genes on human adipogenesis, a second human preadipocyte strain was studied (SGBS) by real-time quantitative PCR (qRT-PCR).A combined epigenetic and transcriptomic approach in differentiated human pre-adipocyte cell line, CHUB-S7, identified that the Human cartilage chitinase 3-like protein 2 ( gene was hypo-methylated and had increased expression in low B12 conditions. Furthermore, there was an approximately 1000-fold increase in expression in the early days of adipocyte differentiation, which paralleled an increase of lipid droplets in differentiated SGBS cells and an increased expression level of markers of mature adipocytes.In summary, we have identified a potential role of the human cartilage chitinase 3-like protein 2 ( in adipocyte function in the presence of low B12 levels.
维生素 B12 具有多种生化功能，包括在一碳循环中生成用于 DNA 甲基化的甲基，以及代谢脂肪酸和氨基酸以通过柠檬酸循环产生能量。我们的研究目的是使用组合的表观基因组和转录组方法来鉴定介导 B12 对脂肪生成作用的新基因。人类前脂肪细胞（CHUB-S7）从细胞接种的那一天开始用一系列 B12（0-500 nM）浓度处理，直到在发现和验证实验中收获，然后在 Illumina HumanMethylation 450Beadchip 上进行全基因组甲基化分析。为了进行转录组分析，在 Illumina HiSeq 2500 上运行 RNA-seq 文库。为了进一步研究任何基因在人类脂肪生成中的表达，通过实时定量 PCR（qRT-PCR）研究了第二种人类前脂肪细胞系（SGBS）。在分化的人类前脂肪细胞系 CHUB-S7 中，采用组合的表观遗传学和转录组学方法，鉴定出人软骨粘蛋白 3 样蛋白 2（基因在低 B12 条件下呈低甲基化状态且表达增加。此外，在脂肪细胞分化的早期，表达增加了约 1000 倍，这与分化的 SGBS 细胞中脂滴的增加以及成熟脂肪细胞标志物表达水平的增加相平行。总之，我们已经确定了人类软骨粘蛋白 3 样蛋白 2（在低 B12 水平存在时，在脂肪细胞功能中可能发挥作用。
