LncRNA-AK137033 inhibits the osteogenic potential of adipose-derived stem cells in diabetic osteoporosis by regulating Wnt signaling pathway via DNA methylation.
LncRNA-AK137033 inhibits the osteogenic potential of adipose-derived stem cells in diabetic osteoporosis by regulating Wnt signaling pathway via DNA methylation.
复制标题
LncRNA-AK137033 通过 DNA 甲基化调节 Wnt 信号通路抑制糖尿病骨质疏松症中脂肪干细胞的成骨潜力
DOI：
10.1111/cpr.13174
复制
发表时间：
2022-01
影响因子：
8.5
通讯作者：
Xiao J
中科院分区：
文献类型：
--
作者：
Peng S;Gao Y;Shi S;Zhao D;Cao H;Fu T;Cai X;Xiao J
基于脂肪干细胞(ASCs)的骨组织工程有望成为治疗糖尿病骨质疏松症(DOP)伴骨缺损的新方法。然而，与对照组ASCs相比，DOP-ASCs的成骨能力降低，增加了DOP患者骨重建的难度。此外，ASCs在高血糖微环境中成骨不良的原因尚不清楚。因此，本研究从表观遗传学角度探讨DOP-ASCs成骨能力下降的分子机制，为DOP伴骨缺损患者的骨修复提供可能的治疗靶点。建立了小鼠DOP动物模型。CON-ASCs和DOP-ASCs分别从CON和DOP小鼠体内分离。利用AK137033小干扰RNA(SiRNA)和AK137033过表达载体调控AK137033在体外培养的CON-ASCs和DOP-ASCs中的表达。用携带shRNA-AK137033和AK137033的慢病毒分别在CON-ASCs和DOP-ASCs体内敲除或过表达AK137033。采用苏木精-伊红(H&E)、Masson氏染色、茜素红和碱性磷酸酶(ALP)染色、显微CT、流式细胞仪、定量聚合酶链式反应(QPCR)、免疫印迹、免疫荧光和亚硫酸盐特异性聚合酶链式反应(BSP)等方法分析ASCs的功能变化。成功建立了DOP小鼠模型。与CON-ASCs相比，DOP-ASCs的AK137033表达、SFRP2启动子区域DNA甲基化水平、Wnt信号通路标志物和成骨分化潜能均降低。体外实验表明，AK137033沉默通过降低SFRP2启动子区域的DNA甲基化水平，抑制了Con-ASCs的Wnt信号通路和成骨能力。此外，AK137033在DOP-ASCs中的过表达挽救了DOP引起的这些变化。此外，在体内也得到了相同的结果。LncRNA-AK137033通过调节SFRP2启动子区域DNA甲基化水平来调节Wnt信号通路，从而抑制DOP-ASCs的成骨潜能。本研究为寻找治疗DOP患者骨缺损的新靶点提供了重要参考。本工作成功构建了糖尿病骨质疏松症(DOP)动物模型。从DOP小鼠体内成功分离培养出DOP-ASCs。体内外研究表明，LncRNA-AK137033通过SFRP2启动子区域DNA甲基化调控Wnt信号通路，从而调节DOP-ASCs的成骨潜能。
Bone tissue engineering based on adipose‐derived stem cells (ASCs) is expected to become a new treatment for diabetic osteoporosis (DOP) patients with bone defects. However, compared with control ASCs (CON‐ASCs), osteogenic potential of DOP‐ASCs is decreased, which increased the difficulty of bone reconstruction in DOP patients. Moreover, the cause of the poor osteogenesis of ASCs in a hyperglycemic microenvironment has not been elucidated. Therefore, this study explored the molecular mechanism of the decline in the osteogenic potential of DOP‐ASCs from the perspective of epigenetics to provide a possible therapeutic target for bone repair in DOP patients with bone defects. An animal model of DOP was established in mice. CON‐ASCs and DOP‐ASCs were isolated from CON and DOP mice, respectively. AK137033 small interfering RNA (SiRNA) and an AK137033 overexpression plasmid were used to regulate the expression of AK137033 in CON‐ASCs and DOP‐ASCs in vitro. Lentiviruses that carried shRNA‐AK137033 or AK137033 cDNA were used to knockdown or overexpress AK137033, respectively, in CON‐ASCs and DOP‐ASCs in vivo. Hematoxylin and eosin (H&E), Masson's, alizarin red, and alkaline phosphatase (ALP) staining, micro‐computed tomography (Micro‐CT), flow cytometry, qPCR, western blotting, immunofluorescence, and bisulfite‐specific PCR (BSP) were used to analyze the functional changes of ASCs. The DOP mouse model was established successfully. Compared with CON‐ASCs, AK137033 expression, the DNA methylation level of the sFrp2 promoter region, Wnt signaling pathway markers, and the osteogenic differentiation potential were decreased in DOP‐ASCs. In vitro experiments showed that AK137033 silencing inhibited the Wnt signaling pathway and osteogenic ability of CON‐ASCs by reducing the DNA methylation level in the sFrp2 promoter region. Additionally, overexpression of AK137033 in DOP‐ASCs rescued these changes caused by DOP. Moreover, the same results were obtained in vivo. LncRNA‐AK137033 inhibits the osteogenic potential of DOP‐ASCs by regulating the Wnt signaling pathway via modulating the DNA methylation level in the sFrp2 promoter region. This study provides an important reference to find new targets for the treatment of bone defects in DOP patients. In this work, the diabetic osteoporosis (DOP) animal model was successfully constructed. DOP‐ASCs were successfully isolated and cultured from DOP mice. In vitro and in vivo studies have demonstrated that LncRNA‐AK137033 regulates the osteogenic potential of DOP‐ASCs by modulating the Wnt signaling pathway via DNA methylation in the sFrp2 promoter region.
登录
查看更多内容
POSTN 的敲低会抑制类固醇引起的骨坏死患者的间充质干细胞的成骨分化。
影响因子：
5.5
作者：
Han L;Gong S;Wang R;Liu S;Wang B;Chen G;Gong T;Xu W
通讯作者：
Xu W
肺腺癌和肺鳞癌中潜在的长非编码 RNA 表达定量特征甲基化的鉴定
影响因子：
3.7
作者：
Wu X;Gao Y;Bu J;Deng L;Zhang P;Chi M;Jiang L;Shi X;Ning S;Wang G
通讯作者：
Wang G
影响因子：
17.1
作者：
Fan J;Lee CS;Kim S;Chen C;Aghaloo T;Lee M
通讯作者：
Lee M
影响因子：
9
作者：
Li M;Cong R;Yang L;Yang L;Zhang Y;Fu Q
通讯作者：
Fu Q
影响因子：
5.8
作者：
Kar, Sumanta;Jasuja, Haneesh;Katti, Kalpana S.
通讯作者：
Katti, Kalpana S.