TGFβ Triggers miR-143/145 Transfer From Smooth Muscle Cells to Endothelial Cells, Thereby Modulating Vessel Stabilization
TGFβ Triggers miR-143/145 Transfer From Smooth Muscle Cells to Endothelial Cells, Thereby Modulating Vessel Stabilization
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DOI：
10.1161/circresaha.116.305178
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发表时间：
2015-05-22
影响因子：
20.1
通讯作者：
Elia, Leonardo
中科院分区：
文献类型：
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作者：
Climent, Montserrat;Quintavalle, Manuela;Elia, Leonardo
基本原理：miR-143/145簇在平滑肌细胞（SMC）中高度表达，在那里它调节表型转换和血管稳态。目的：研究miR-143和miR-145在内皮细胞（EC）中的作用。方法：采用不同培养条件下的SMC和EC共培养，以及完整的血管，研究miR-143和miR-145在不同细胞类型间的转移。用荧光miRNA转导的共培养细胞的成像表明，miRNA转移涉及称为隧道纳米管的膜突起。此外，我们还发现，由于特异性转化生长因子β（TGF β）抑制剂（SB 431542）和针对TGF β RII的shRNA均抑制了miR-143/145从SMC向EC的传递，因此miRNA的传递受到TGF β途径的调节。此外，当在基质胶上培养时，miR-143和miR-145通过降低EC的增殖指数及其形成血管样结构的能力来调节血管生成。我们还鉴定了对内皮细胞血管生成潜力至关重要的己糖激酶II（HKII）和整合素β 8（ITG β 8）-2基因，分别作为miR-143和miR-145的靶基因。这些基因的抑制调节EC表型，类似于miR-143和miR-145在EC中的过表达。这些研究结果证实了离体和体内的方法，其中，TGF β和血管应力，分别触发miR-143/145从SMC转移到ECs.Conclusions：我们的研究结果表明，miR-143和miR-145作为SMC和EC之间的通信分子，以调节血管生成和血管稳定性能的EC。
Rationale: The miR-143/145 cluster is highly expressed in smooth muscle cells (SMCs), where it regulates phenotypic switch and vascular homeostasis. Whether it plays a role in neighboring endothelial cells (ECs) is still unknown.Objective: To determine whether SMCs control EC functions through passage of miR-143 and miR-145.Methods and Results: We used cocultures of SMCs and ECs under different conditions, as well as intact vessels to assess the transfer of miR-143 and miR-145 from one cell type to another. Imaging of cocultured cells transduced with fluorescent miRNAs suggested that miRNA transfer involves membrane protrusions known as tunneling nanotubes. Furthermore, we show that miRNA passage is modulated by the transforming growth factor (TGF) beta pathway because both a specific transforming growth factor-beta (TGF beta) inhibitor (SB431542) and an shRNA against TGF beta RII suppressed the passage of miR-143/145 from SMCs to ECs. Moreover, miR-143 and miR-145 modulated angiogenesis by reducing the proliferation index of ECs and their capacity to form vessel-like structures when cultured on matrigel. We also identified hexokinase II (HKII) and integrin beta 8 (ITG beta 8)-2 genes essential for the angiogenic potential of ECs-as targets of miR-143 and miR-145, respectively. The inhibition of these genes modulated EC phenotype, similarly to miR-143 and miR-145 overexpression in ECs. These findings were confirmed by ex vivo and in vivo approaches, in which it was shown that TGF beta and vessel stress, respectively, triggered miR-143/145 transfer from SMCs to ECs.Conclusions: Our results demonstrate that miR-143 and miR-145 act as communication molecules between SMCs and ECs to modulate the angiogenic and vessel stabilization properties of ECs.