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Improving the Angiogenic Potential of EPCs via Engineering with Synthetic Modified mRNAs
Molecular Therapy - Nucleic Acids ( IF 8.8 ) Pub Date : 2018-09-13 , DOI: 10.1016/j.omtn.2018.09.005
Heidrun Steinle , Sonia Golombek , Andreas Behring , Christian Schlensak , Hans Peter Wendel , Meltem Avci-Adali

The application of endothelial progenitor cells (EPCs) for the revascularization of ischemic tissues, such as after myocardial infarction, stroke, and acute limb ischemia, has a huge clinical potential. However, the low retention and engraftment of EPCs as well as the poor survival of migrated stem cells in ischemic tissues still hamper the successful clinical application. Thus, in this study, we engineered, for the first time, murine EPCs with synthetic mRNAs to transiently produce proangiogenic factors vascular endothelial growth factor-A (VEGF-A), stromal cell-derived factor-1α (SDF-1α), and angiopoietin-1 (ANG-1). After the transfection of cells with synthetic mRNAs, significantly increased VEGF-A, SDF-1α, and ANG-1 protein levels were detected compared to untreated EPCs. Thereby, mRNA-engineered EPCs showed significantly increased chemotactic activity versus untreated EPCs and resulted in significantly improved attraction of EPCs. Furthermore, ANG-1 mRNA-transfected EPCs displayed a strong wound-healing capacity. Already after 12 hr, 94% of the created wound area in the scratch assay was closed compared to approximately 45% by untreated EPCs. Moreover, the transfection of EPCs with ANG-1 or SDF-1α mRNA also significantly improved the in vitro tube formation capacity; however, the strongest effect could be detected with EPCs simultaneously transfected with VEGF-A, SDF-1α, and ANG-1 mRNA. In the in vivo chicken chorioallantoic membrane (CAM) assay, EPCs transfected with ANG-1 mRNA revealed the strongest angiogenetic potential with significantly elevated vessel density and total vessel network length. In conclusion, this study demonstrated that EPCs can be successfully engineered with synthetic mRNAs encoding proangiogenic factors to improve their therapeutic angiogenetic potential in patients experiencing chronic or acute ischemic disease.



中文翻译:

通过合成修饰的mRNA工程提高EPC的血管生成潜能

内皮祖细胞(EPC)在缺血性组织的血运重建中的应用,例如在心肌梗塞,中风和急性肢体缺血后,具有巨大的临床潜力。然而,EPC的低保留和植入以及在缺血组织中迁移的干细胞的不良存活仍然阻碍了成功的临床应用。因此,在这项研究中,我们首次设计了具有合成mRNA的鼠EPC,以瞬时产生促血管生成因子,血管内皮生长因子-A(VEGF-A),基质细胞衍生因子-1α(SDF-1α)和血管生成素-1(ANG-1)。用合成的mRNA转染细胞后,与未处理的EPC相比,检测到VEGF-A,SDF-1α和ANG-1蛋白水平显着增加。从而,与未经处理的EPC相比,mRNA工程化的EPC表现出显着提高的趋化活性,并导致EPC的吸引力显着提高。此外,ANG-1 mRNA转染的EPCs表现出很强的伤口愈合能力。在12小时之后,已经关闭了刮擦试验中94%的创口面积,相比之下,未经处理的EPC约为45%。此外,用ANG-1或SDF-1αmRNA转染EPCs还可显着改善体外管形成能力;然而,用同时转染VEGF-A,SDF-1α和ANG-1 mRNA的EPC可以检测到最强的作用。在体内鸡绒膜尿囊膜(CAM)分析中,转染ANG-1 mRNA的EPCs显示出最强的血管生成潜能,血管密度和总血管网络长度明显增加。总之,这项研究表明,EPCs可以成功地用编码促血管生成因子的合成mRNA进行工程改造,以改善患有慢性或急性缺血性疾病的患者的治疗性血管生成潜力。

更新日期:2018-09-13
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