Alkbh1-mediated DNA N6-methyladenine modification regulates bone marrow mesenchymal stem cell fate during skeletal aging.
Alkbh1-mediated DNA N6-methyladenine modification regulates bone marrow mesenchymal stem cell fate during skeletal aging.
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Alkbh 1介导的DNA N6-甲基腺嘌呤修饰调节骨骼衰老过程中骨髓间充质干细胞的命运
DOI：
10.1111/cpr.13178
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发表时间：
2022-03
影响因子：
8.5
通讯作者：
Wang M
中科院分区：
文献类型：
--
作者：
Cai GP;Liu YL;Luo LP;Xiao Y;Jiang TJ;Yuan J;Wang M
DNA N6-甲基腺嘌呤（N6 - mA）去甲基化酶Alkbh1参与调控间充质干细胞（MSCs）的成骨分化及血管钙化。然而，Alkbh1在骨代谢中的作用仍不明确。
本研究利用骨髓间充质干细胞（BMSCs）特异性Alkbh1基因敲除小鼠，探究Alkbh1在骨代谢中的作用。采用蛋白质免疫印迹法（Western blot）、实时荧光定量聚合酶链反应（qRT - PCR）及免疫荧光染色法检测Alkbh1或视神经蛋白（optn）的表达。运用显微计算机断层扫描（Micro - CT）、组织形态计量分析及钙黄绿素双标实验评估骨表型。通过细胞染色及qRT - PCR检测BMSCs的成骨或成脂分化情况。采用斑点印迹法检测基因组DNA中N6 - mA水平。利用染色质免疫沉淀（Chip）实验鉴定Alkbh1的关键靶点。使用携带Alkbh1的腺相关病毒在老年小鼠中过表达Alkbh1。
衰老过程中，BMSCs中Alkbh1的表达下降。Alkbh1基因敲除促进BMSCs的成脂分化，同时抑制其成骨分化。BMSCs特异性Alkbh1基因敲除小鼠表现出骨量减少及骨髓脂肪增多。从机制上看，我们确定optn是Alkbh1介导的DNA m6A修饰调控BMSCs命运的下游靶点。Alkbh1过表达可减轻老年小鼠的骨质流失及骨髓脂肪积累。
我们的研究结果表明，Alkbh1在骨骼衰老过程中调控BMSCs的命运及骨 - 脂肪平衡，为骨质疏松症的治疗提供了一个潜在靶点。
我们的研究结果揭示，DNA N6 - 甲基腺嘌呤去甲基化酶Alkbh1在衰老过程中调控骨髓间充质干细胞的命运。Alkbh1缺失抑制骨形成并促进骨髓脂肪积累。从机制上看，我们确定optn是Alkbh1介导的DNA m6A修饰调控BMSCs命运的下游靶点。
DNA N6‐methyladenine (N6‐mA) demethylase Alkbh1 participates in regulating osteogenic differentiation of mesenchymal stem cell (MSCs) and vascular calcification. However, the role of Alkbh1 in bone metabolism remains unclear. Bone marrow mesenchymal stem cells (BMSCs)‐specific Alkbh1 knockout mice were used to investigate the role of Alkbh1 in bone metabolism. Western blot, qRT‐PCR, and immunofluorescent staining were used to evaluate the expression of Alkbh1 or optineurin (optn). Micro‐CT, histomorphometric analysis, and calcein double‐labeling assay were used to evaluate bone phenotypes. Cell staining and qRT‐PCR were used to evaluate the osteogenic or adipogenic differentiation of BMSCs. Dot blotting was used to detect the level of N6‐mA in genomic DNA. Chromatin immunoprecipitation (Chip) assays were used to identify critical targets of Alkbh1. Alkbh1 adeno‐associated virus was used to overexpress Alkbh1 in aged mice. Alkbh1 expression in BMSCs declined during aging. Knockout of Alkbh1 promoted adipogenic differentiation of BMSCs while inhibited osteogenic differentiation. BMSC‐specific Alkbh1 knockout mice exhibited reduced bone mass and increased marrow adiposity. Mechanistically, we identified optn as the downstream target through which Alkbh1‐mediated DNA m6A modification regulated BMSCs fate. Overexpression of Alkbh1 attenuated bone loss and marrow fat accumulation in aged mice. Our findings demonstrated that Alkbh1 regulated BMSCs fate and bone‐fat balance during skeletal aging and provided a potential target for the treatment of osteoporosis. Our findings revealed that DNA N6‐methyladenine demethylase Alkbh1 regulates bone marrow mesenchymal stem cell fate during aging. Loss of Alkbh1 inhibited bone formation and promoted marrow fat accumulation. Mechanistically, we identified optn as the downstream target through which Alkbh1‐mediated DNA m6A modification regulated BMSCs fate.
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影响因子：
29
作者：
Forte D;García-Fernández M;Sánchez-Aguilera A;Stavropoulou V;Fielding C;Martín-Pérez D;López JA;Costa ASH;Tronci L;Nikitopoulou E;Barber M;Gallipoli P;Marando L;Fernández de Castillejo CL;Tzankov A;Dietmann S;Cavo M;Catani L;Curti A;Vázquez J;Frezza C;Huntly BJ;Schwaller J;Méndez-Ferrer S
通讯作者：
Méndez-Ferrer S
全基因组关联研究将CSF1，OPTN和TNFRSF11A的变体确定为Paget骨骼疾病的遗传危险因素。
影响因子：
30.8
作者：
通讯作者：
--
DOI：
10.1016/j.bbrc.2017.10.158
发表时间：
2018-01-01
影响因子：
3.1
作者：
Müller TA;Struble SL;Meek K;Hausinger RP
通讯作者：
Hausinger RP
影响因子：
3.7
作者：
Benisch P;Schilling T;Klein-Hitpass L;Frey SP;Seefried L;Raaijmakers N;Krug M;Regensburger M;Zeck S;Schinke T;Amling M;Ebert R;Jakob F
通讯作者：
Jakob F
ALKBH1是参与神经分化的组蛋白H2A双加氧酶。
影响因子：
5.2
作者：
Ougland, Rune;Lando, David;Jonson, Ida;Dahl, John A.;Moen, Marivi Nabong;Nordstrand, Line M.;Rognes, Torbjorn;Lee, Jeannie T.;Klungland, Arne;Kouzarides, Tony;Larsen, Elisabeth
通讯作者：
Larsen, Elisabeth