Periodontitis-compromised dental pulp stem cells secrete extracellular vesicles carrying miRNA-378a promote local angiogenesis by targeting Sufu to activate the Hedgehog/Gli1 signalling.
Periodontitis-compromised dental pulp stem cells secrete extracellular vesicles carrying miRNA-378a promote local angiogenesis by targeting Sufu to activate the Hedgehog/Gli1 signalling.
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牙周炎受损的牙髓干细胞分泌携带 miRNA-378a 的细胞外囊泡，通过靶向 Sufu 激活 Hedgehog/Gli1 信号，促进局部血管生成
DOI：
10.1111/cpr.13026
复制
发表时间：
2021-05
影响因子：
8.5
通讯作者：
Chen FM
中科院分区：
文献类型：
--
作者：
Zhou H;Li X;Wu RX;He XT;An Y;Xu XY;Sun HH;Wu LA;Chen FM
以前，我们的研究表明，与健康DPSC（H-EV）相比，牙周炎受损牙髓干细胞（P-EV）分泌的细胞外囊泡具有强大的促血管生成潜力，但其潜在机制仍然未知。在这里，通过Agilent miRNA微阵列分析鉴定了在P-EV中特异性发现的循环microRNA（miRNA）（与H-EV相比），并通过细胞转染和RNA干扰方法证实了候选miRNA在P-EV增强细胞血管生成中的作用。接着，通过荧光素酶报告基因测定评估候选miRNA与其靶基因之间的直接结合亲和力。建立CCK-8、transwell/scratch伤口愈合和管形成试验，以研究内皮细胞（EC）的增殖、迁移和管形成能力。Western blot检测Hedgehog/Gli 1信号通路组分和血管生成相关因子的蛋白水平。发现血管生成相关的miRNA miR-378 a在P-EV中富集，并证实了其在P-EV增强细胞血管生成中的作用，其中Sufu被鉴定为miR-378 a的下游靶基因。在功能上，沉默腐乳刺激EC增殖，迁移和管形成激活Hedgehog/Gli 1信号。此外，我们发现与P-EV一起孵育使得含有P-EV的miR-378 a能够传递到EC。随后，P-EV-介导的miR-378 a传递显着影响EC中Sufu、Gli 1和血管内皮生长因子的表达。这些数据表明，携带miR-378 a的P-EV通过下调Sufu以激活Hedgehog/Gli 1信号通路来促进EC血管生成。我们的研究结果揭示了EV衍生的miR-378 a在细胞血管生成中的关键作用，因此为修饰干细胞及其分泌的EV提供了新的靶点，以增强血管再生潜力。携带miR-378 a的P-EV通过下调Sufu以激活Hedgehog/Gli 1信号通路来促进EC的血管生成。
Previously, our investigations demonstrated robust pro‐angiogenic potentials of extracellular vesicles secreted by periodontitis‐compromised dental pulp stem cells (P‐EVs) when compared to those from healthy DPSCs (H‐EVs), but the underlying mechanism remains unknown. Here, circulating microRNAs (miRNAs) specifically found in P‐EVs (compared with H‐EVs) were identified by Agilent miRNA microarray analysis, and the roles of the candidate miRNA in P‐EV‐enhanced cell angiogenesis were confirmed by cell transfection and RNA interference methods. Next, the direct binding affinity between the candidate miRNA and its target gene was evaluated by luciferase reporter assay. CCK‐8, transwell/scratch wound healing and tube formation assays were established to investigate the proliferation, migration, and tube formation abilities of endothelial cells (ECs). Western blot was employed to measure the protein levels of Hedgehog/Gli1 signalling pathway components and angiogenesis‐related factors. The angiogenesis‐related miRNA miR‐378a was found to be enriched in P‐EVs, and its role in P‐EV‐enhanced cell angiogenesis was confirmed, wherein Sufu was identified as a downstream target gene of miR‐378a. Functionally, silencing of Sufu stimulated EC proliferation, migration and tube formation by activating Hedgehog/Gli1 signalling. Further, we found that incubation with P‐EVs enabled the transmission of P‐EV‐contained miR‐378a to ECs. Subsequently, the expressions of Sufu, Gli1 and vascular endothelial growth factor in ECs were significantly influenced by P‐EV‐mediated miR‐378a transmission. These data suggest that P‐EVs carrying miR‐378a promote EC angiogenesis by downregulating Sufu to activate the Hedgehog/Gli1 signalling pathway. Our findings reveal a crucial role for EV‐derived miR‐378a in cell angiogenesis and hence offer a new target for modifying stem cells and their secreted EVs to enhance vessel regenerative potential. P‐EVs carrying miR‐378a promote the angiogenesis of ECs by down‐regulating Sufu to activate the Hedgehog/Gli1 signalling pathway.
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DOI：
10.1126/science.aau6977
发表时间：
2020-02-07
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Kalluri R;LeBleu VS
通讯作者：
LeBleu VS
DOI：
10.1073/pnas.1815447116
发表时间：
2019-01-29
影响因子：
11.1
作者：
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通讯作者：
Mauck, Robert L.
DOI：
10.1073/pnas.1808855115
发表时间：
2018-11-27
影响因子：
11.1
作者：
Castano, Carlos;Kalko, Susana;Parrizas, Marcelina
通讯作者：
Parrizas, Marcelina
影响因子：
9.8
作者：
Nagase, Takashi;Nagase, Miki;Fujita, Toshiro
通讯作者：
Fujita, Toshiro
影响因子：
5.5
作者：
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Wang Z