MicroRNA-205 mediates endothelial progenitor functions in distraction osteogenesis by targeting the transcription regulator NOTCH2.
MicroRNA-205 mediates endothelial progenitor functions in distraction osteogenesis by targeting the transcription regulator NOTCH2.
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MicroRNA-205通过靶向转录调节因子NOTCH2介导牵张成骨中的内皮祖细胞功能
DOI：
10.1186/s13287-021-02150-x
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发表时间：
2021-02-03
影响因子：
7.5
通讯作者：
Zhou N
中科院分区：
文献类型：
--
作者：
Jiang W;Zhu P;Zhang T;Liao F;Yu Y;Liu Y;Shen H;Zhao Z;Huang X;Zhou N
牵张成骨（DO）是一种非常有效的重建性骨再生形式，但其临床应用受到骨整合发生所需时间延长的限制。因此，了解DO的机制基础和缩短巩固期代表了改善该手术临床实用性的有希望的方法。建立下颌骨DO（MDO）犬模型，通过小分子RNA测序鉴定相关分子靶基因。通过生物信息学鉴定推定的miRNA靶基因，并通过qPCR、Western印迹和双荧光素酶报告基因测定进行确认。收集外周血样品以分离血清和内皮集落形成细胞（ECFC），以测量miR-205、N 0 TCH 2和血管生成细胞因子表达水平。然后使用慢病毒构建体来抑制或过表达分离的ECFC中的miR-205和NOTCH 2，之后在迁移、伤口愈合、增殖、管形成和鸡绒毛尿囊膜（CAM）测定中评价这些细胞的血管生成活性。转染AutoblastECFC以敲低miR-205并直接注射到牵张愈伤组织中。术后第14、28、35、42天，通过CBCT评价骨密度，收集骨痂样品并通过组织学染色评价以分析骨再生和重塑。MiR-205被鉴定为在MDO愈伤组织样品中最显著下调的miRNA之一。在经历MDO的动物的DO-ECFC和血清中也观察到miR-205的下调。抑制miR-205显著增强血管生成，而过表达miR-205在体外具有相反的效果。重要的是，NOTCH 2是骨血管生成中的独特调节因子，被鉴定为miR-205靶基因。与这种调节关系一致，敲低NOTCH 2抑制血管生成，并且用miR-205抑制剂慢病毒转导足以挽救血管生成活性。当将其中miR-205已被抑制的ECFC移植到MDO愈伤组织中时，这显著增强了体内骨生成和重塑。MiR-205是MDO过程的重要调节剂，抑制这种miRNA可以加速MDO相关的矿化。总的来说，这些结果提供了新的见解，这种程序的机制基础，突出治疗性临床干预的潜在目标。在线版本包含补充材料，可通过10.1186/s13287-021-02150-x获得。
Distraction osteogenesis (DO) is a highly efficacious form of reconstructive bone regeneration, but its clinical utility is limited by the prolonged period required for bone consolidation to occur. Understanding the mechanistic basis for DO and shortening this consolidation phase thus represent promising approaches to improving the clinical utility of this procedure. A mandibular DO (MDO) canine model was established, after which small RNA sequencing was performed to identify relevant molecular targets genes. Putative miRNA target genes were identified through bioinformatics and confirmed through qPCR, Western blotting, and dual-luciferase reporter assays. Peripheral blood samples were collected to isolate serum and endothelial colony-forming cells (ECFCs) in order to measure miR-205, NOTCH2, and angiogenic cytokines expression levels. Lentiviral constructs were then used to inhibit or overexpress miR-205 and NOTCH2 in isolated ECFCs, after which the angiogenic activity of these cells was evaluated in migration, wound healing, proliferation, tube formation, and chick chorioallantoic membrane (CAM) assay. Autologous ECFCs transfected to knockdown miR-205 and were injected directly into the distraction callus. On days 14, 28, 35 and 42 after surgery, bone density was evaluated via CBCT, and callus samples were collected and evaluated via histological staining to analyze bone regeneration and remodeling. MiR-205 was identified as being one of the miRNAs that was most significantly downregulated in MDO callus samples. Downregulation of miR-205 was also observed in DO-ECFCs and serum of animals undergoing MDO. Inhibiting miR-205 markedly enhanced angiogenesis, whereas overexpressing miR-205 had the opposite effect in vitro. Importantly, NOTCH2, which is a unique regulator in bone angiogenesis, was identified as a miR-205 target gene. Consistent with this regulatory relationship, knocking down NOTCH2 suppressed angiogenesis, and transduction with a miR-205 inhibitor lentivirus was sufficient to rescue angiogenic activity. When ECFCs in which miR-205 had been inhibited were transplanted into the MDO callus, this significantly bolstered osteogenesis, and remodeling in vivo. MiR-205 is a significant regulator of the MDO process, and inhibiting this miRNA can accelerate MDO-related mineralization. Overall, these results offer new insights into the mechanistic basis for this procedure, highlighting potential targets for therapeutic clinical intervention. The online version contains supplementary material available at 10.1186/s13287-021-02150-x.
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影响因子：
6
作者：
Medina RJ;Barber CL;Sabatier F;Dignat-George F;Melero-Martin JM;Khosrotehrani K;Ohneda O;Randi AM;Chan JKY;Yamaguchi T;Van Hinsbergh VWM;Yoder MC;Stitt AW
通讯作者：
Stitt AW
影响因子：
12.4
作者：
He, Liuqing;Zhu, Wei;Wu, Xiaoying
通讯作者：
Wu, Xiaoying
影响因子：
4.1
作者：
Lee, Dong Yeon;Cho, Tae-Joon;Choi, In Ho
通讯作者：
Choi, In Ho
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影响因子：
7.5
作者：
Jia, Yachao;Zhu, Yu;Chai, Yimin
通讯作者：
Chai, Yimin
影响因子：
64.8
作者：
Kusumbe AP;Ramasamy SK;Adams RH
通讯作者：
Adams RH