Laboratory of Molecular Signaling and Stem Cells Therapy, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Capital Medical University School of Stomatology, Beijing, China.
Department of Dental Implant Center, Beijing Stomatological Hospital, School of Stomatology, Capital Medical University, Beijing, China.
Int J Oral Sci. 2023 Jan 12;15(1):6. doi: 10.1038/s41368-022-00202-3.
The failure rate of dental implantation in patients with well-controlled type 2 diabetes mellitus (T2DM) is higher than that in non-diabetic patients. This due, in part, to the impaired function of bone marrow mesenchymal stem cells (BMSCs) from the jawbone marrow of T2DM patients (DM-BMSCs), limiting implant osseointegration. RNA N6-methyladenine (m6A) is important for BMSC function and diabetes regulation. However, it remains unclear how to best regulate m6A modifications in DM-BMSCs to enhance function. Based on the "m6A site methylation stoichiometry" of m6A single nucleotide arrays, we identified 834 differential m6A-methylated genes in DM-BMSCs compared with normal-BMSCs (N-BMSCs), including 43 and 790 m6A hypermethylated and hypomethylated genes, respectively, and 1 gene containing hyper- and hypomethylated m6A sites. Differential m6A hypermethylated sites were primarily distributed in the coding sequence, while hypomethylated sites were mainly in the 3'-untranslated region. The largest and smallest proportions of m6A-methylated genes were on chromosome 1 and 21, respectively. MazF-PCR and real-time RT-PCR results for the validation of erythrocyte membrane protein band 4.1 like 3, activity-dependent neuroprotector homeobox (ADNP), growth differentiation factor 11 (GDF11), and regulator of G protein signalling 2 agree with m6A single nucleotide array results; ADNP and GDF11 mRNA expression decreased in DM-BMSCs. Furthermore, gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses suggested that most of these genes were enriched in metabolic processes. This study reveals the differential m6A sites of DM-BMSCs compared with N-BMSCs and identifies candidate target genes to enhance BMSC function and improve implantation success in T2DM patients.
患有 2 型糖尿病（T2DM）的患者中，牙种植体的失败率高于非糖尿病患者。部分原因是 T2DM 患者颌骨骨髓间充质干细胞（BMSCs，DM-BMSCs）的功能受损，限制了种植体的骨整合。RNA N6-甲基腺苷（m6A）对 BMSC 功能和糖尿病调节很重要。然而，目前尚不清楚如何最好地调节 DM-BMSCs 中的 m6A 修饰以增强功能。基于 m6A 单核苷酸阵列的“m6A 位点甲基化化学计量”，我们发现与正常 BMSCs（N-BMSCs）相比，DM-BMSCs 中有 834 个差异 m6A 甲基化基因，分别有 43 个和 790 个 m6A 高甲基化和低甲基化基因，以及 1 个含有高甲基化和低甲基化 m6A 位点的基因。差异 m6A 高甲基化位点主要分布在编码序列中，而低甲基化位点主要位于 3'-非翻译区。m6A 甲基化基因的最大和最小比例分别位于 1 号和 21 号染色体上。红细胞膜蛋白带 4.1 样 3、活性依赖性神经保护同源框（ADNP）、生长分化因子 11（GDF11）和 G 蛋白信号调节因子 2 的 MazF-PCR 和实时 RT-PCR 结果验证与 m6A 单核苷酸阵列结果一致；ADNP 和 GDF11 的 mRNA 表达在 DM-BMSCs 中降低。此外，基因本体论和京都基因与基因组百科全书分析表明，这些基因大多富集在代谢过程中。本研究揭示了 DM-BMSCs 与 N-BMSCs 相比的差异 m6A 位点，并确定了候选靶基因，以增强 BMSC 功能，提高 T2DM 患者的种植体成功率。
