从肾主骨生髓探讨骨质疏松症中m6A去甲基化转移酶FTO介导成骨的分子机制研究

N6-甲基腺苷（m6A)是真核mRNA中最丰富的表观遗传修饰，广泛分布于基因转录区，能在转录后水平调节基因表达，最新研究发现m6A修饰在骨骼发育和代谢中起重要作用。脂肪和肥胖相关蛋白（FTO）是已知能在体内外调控m6A修饰的RNA去甲基化酶，我们前期实验发现沉默FTO可显著抑制间充质干细胞（MSCs）成骨分化，但FTO-m6A修饰对MSCs谱系分化、骨质疏松发生、发展作用机制仍需深入探讨。故本项目拟：①以FTO基因敲除小鼠为工具，建立MSCs成骨分化模型，通过RNA甲基化测序、转录组测序技术，研究FTO-m6A修饰与MSCs骨形成基因的关联；②借助骨质疏松动物模型及临床样本，采用CRISPR基因敲除和慢病毒基因超表达技术，研究FTO-m6A修饰在补肾法治疗骨质疏松中的作用，探明RNA去甲基化修饰在骨形成中的分子作用机理，及与骨质疏松的关联性，为中医药精准防治骨质疏松提供新思路和理论依据。

N6-methyladenosine (m6A) is the most abundant epigenetic modification in eukaryotic mRNA. It is widely distributed in gene transcription regions and can regulate gene expression at the post-transcription level. Recent studies have found that m6A modification plays a role in bone development and metabolism. Important role. Fat and obesity-related protein (FTO) are RNA demethylases known to regulate m6A modification in vitro and in vivo. Our previous experiments found that silencing FTO can significantly inhibit osteogenic differentiation of mesenchymal stem cells (MSCs), but FTO The mechanism of -m6A modification on MSCs lineage differentiation, osteoporosis and development needs to be further explored. Therefore, this project intends to: ①establish a model of MSCs osteogenic differentiation using FTO knockout mice as a tool, and study the association between FTO-m6A modification and MSCs bone formation genes through MeRIP-seq and RNA-seq technologies; ②with the help of bone In animal models and clinical samples of osteoporosis, gene silencing and overexpression techniques were used to study the role of FTO-m6A modification in the treatment of osteoporosis with kidney-reinforcing method, and to investigate the molecular mechanism of RNA demethylation modification in bone formation, and The relevance of osteoporosis provides new ideas and theoretical basis for the accurate prevention and treatment of osteoporosis in traditional Chinese medicine.