Elastin is one of the most important and abundant extracellular matrix (ECM) proteins that provide elasticity and resilience to tissues and organs, including vascular walls, ligaments, skin, and lung. Besides hereditary diseases, such as Williams-Beuren syndrome (WBS), which results in reduced elastin synthesis, injuries, aging, or acquired diseases can lead to the degradation of existing elastin fibers. Thus, the de novo synthesis of elastin is required in several medical conditions to restore the elasticity of affected tissues. Here, we applied synthetic modified mRNA encoding tropoelastin (TE) for the de novo synthesis of elastin and determined the mRNA-mediated elastin synthesis in cells, as well as ex vivo in porcine skin. EA.hy926 cells, human fibroblasts, and mesenchymal stem cells (MSCs) isolated from a patient with WBS were transfected with 2.5 μg TE mRNA. After 24 hr, the production of elastin was analyzed by Fastin assay and dot blot analyses. Compared with untreated cells, significantly enhanced elastin amounts were detected in TE mRNA transfected cells. The delivered synthetic TE mRNA was even able to significantly increase the elastin production in elastin-deficient MSCs. In porcine skin, approximately 20% higher elastin amount was detected after the intradermal delivery of synthetic mRNA by microinjection. In this study, we demonstrated the successful applicability of synthetic TE encoding mRNA to produce elastin in elastin-deficient cells as well as in skin. Thus, this auspicious mRNA-based integration-free method has a huge potential in the field of regenerative medicine to induce de novo elastin synthesis, e.g., in skin, blood vessels, or alveoli.

弹性蛋白是最重要和最丰富的细胞外基质 (ECM) 蛋白之一，为组织和器官（包括血管壁、韧带、皮肤和肺）提供弹性和复原力。除了导致弹性蛋白合成减少的威廉姆斯-博伊伦综合征 (WBS) 等遗传性疾病之外，损伤、衰老或后天性疾病也会导致现有弹性蛋白纤维的降解。因此，在多种医疗状况下需要从头合成弹性蛋白来恢复受影响组织的弹性。在这里，我们应用合成修饰的编码弹性蛋白原 (TE) 的 mRNA 来从头合成弹性蛋白，并确定了细胞中以及猪皮肤中 mRNA 介导的弹性蛋白合成。使用 2.5 μg TE mRNA 转染从 WBS 患者分离的 EA.hy926 细胞、人成纤维细胞和间充质干细胞 (MSC)。24小时后，通过Fastin测定和斑点印迹分析来分析弹性蛋白的产生。与未处理的细胞相比，TE mRNA 转染细胞中的弹性蛋白含量显着增加。递送的合成 TE mRNA 甚至能够显着增加缺乏弹性蛋白的 MSC 中的弹性蛋白产量。在猪皮肤中，通过显微注射皮内递送合成 mRNA 后，检测到弹性蛋白含量增加了约 20%。在这项研究中，我们证明了合成 TE 编码 mRNA 在弹性蛋白缺陷细胞和皮肤中产生弹性蛋白的成功应用。因此，这种基于 mRNA 的免整合方法在再生医学领域具有巨大的潜力，可以诱导皮肤、血管或肺泡中的弹性蛋白从头合成。