Fernandes Hugo, Zonnari Alessandra, Abreu Ricardo, Aday Sezin, Barão Marta, Albino Inês, Lino Miguel, Branco Ana, Seabra Cátia, Barata Tânia, Leal Ermelindo C, Tralhão José Guilherme, Gonçalves Lino, de Jong Alwin, Peters Hendrika A B, de Vries Margreet R, da Costa Martins Paula, Quax Paul H A, Ferreira Lino
CNC-Center for Neuroscience and Cell Biology, CIBB-Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, UC, Biotech Parque Tecnológico de Cantanhede, 3060-197 Coimbra, Portugal.
Faculty of Medicine, University Coimbra, 3000-548 Coimbra, Portugal.
Mol Ther Nucleic Acids. 2022 Mar 19;28:307-327. doi: 10.1016/j.omtn.2022.03.018. eCollection 2022 Jun 14.
Endothelial cell (EC) activity is essential for tissue regeneration in several (patho)physiological contexts. However, our capacity to deliver biomolecules capable of controlling EC fate is relatively limited. Here, we screened a library of microRNA (miR) mimics and identified 25 miRs capable of enhancing the survival of ECs exposed to ischemia-mimicking conditions. , we showed that miR-425-5p, one of the hits, was able to enhance EC survival and migration. , using a mouse Matrigel plug assay, we showed that ECs transfected with miR-425-5p displayed enhanced survival compared with scramble-transfected ECs. Mechanistically, we showed that miR-425-5p modulated the PTEN/PI3K/AKT pathway and inhibition of miR-425-5p target genes (, , , and ) phenocopied the pro-survival. For the delivery of miR-425-5p, we modulated small extracellular vesicles (sEVs) with miR-425-5p and showed, , that miR-425-5p-modulated sEVs were (1) capable of enhancing the survival of ECs exposed to ischemia-mimic conditions, and (2) efficiently internalized by skin cells. Finally, using a streptozotocin-induced diabetic wound healing mouse model, we showed that, compared with miR-scrambled-modulated sEVs, topical administration of miR-425-5p-modulated sEVs significantly enhanced wound healing, a process mediated by enhanced vascularization and skin re-epithelialization.
内皮细胞（EC）活性在多种（病理）生理环境下对组织再生至关重要。然而，我们递送能够控制EC命运的生物分子的能力相对有限。在此，我们筛选了一个微小RNA（miR）模拟物文库，并鉴定出25种能够增强暴露于模拟缺血条件下的EC存活的miR。我们发现，其中一种命中的miR-425-5p能够增强EC的存活和迁移。此外，通过小鼠基质胶栓塞试验，我们发现与乱序转染的EC相比，用miR-425-5p转染的EC显示出增强的存活能力。从机制上讲，我们发现miR-425-5p调节PTEN/PI3K/AKT途径，并且抑制miR-425-5p靶基因（如 、 、 和 ）可模拟其促存活作用。为了递送miR-425-5p，我们用miR-425-5p调节小细胞外囊泡（sEV），并表明，miR-425-5p调节的sEV能够（1）增强暴露于模拟缺血条件下的EC的存活，以及（2）被皮肤细胞有效内化。最后，使用链脲佐菌素诱导的糖尿病伤口愈合小鼠模型，我们发现，与miR乱序调节的sEV相比，局部施用miR-425-5p调节的sEV显著增强了伤口愈合，这一过程由增强的血管生成和皮肤再上皮化介导。
