Department of Convergence Medicine and Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea.
Department of Convergence Medicine and Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea; ConveRgence mEDIcine research cenTer (CREDIT), Asan Institute for Life Sciences, Asan Medical Center, Seoul, Republic of Korea.
Microvasc Res. 2024 Sep;155:104698. doi: 10.1016/j.mvr.2024.104698. Epub 2024 May 25.
Angiogenesis is mainly regulated by the delivery of VEGF-dependent signaling to cells. However, the angiogenesis mechanism regulated by VEGF-induced miRNA is still not understood. After VEGF treatment in HUVECs, we screened the changed miRNAs through small-RNA sequencing and found VEGF-induced miR-4701-3p. Furthermore, the GFP reporter gene was used to reveal that TOB2 expression was regulated by miR-4701-3p, and it was found that TOB2 and miR-4701-3p modulation could cause angiogenesis in an in-vitro angiogenic assay. Through the luciferase assay, it was confirmed that the activation of the angiogenic transcription factor MEF2 was regulated by the suppression and overexpression of TOB2 and miR-4701-3p. As a result, MEF2 downstream gene mRNAs that induce angiogenic function were regulated. We used the NCBI GEO datasets to reveal that the expression of TOB2 and MEF2 was significantly changed in cardiovascular disease. Finally, it was confirmed that the expression of circulating miR-4701-3p in the blood of myocardial infarction patients was remarkably increased. In patients with myocardial infarction, circulating miR-4701-3p was increased regardless of age, BMI, and sex, and showed high AUC levels in specificity and sensitivity analysis (AUROC) (AUC = 0.8451, 95 % CI 0.78-0.90). Our data showed TOB2-mediated modulation of MEF2 and its angiogenesis by VEGF-induced miR-4701-3p in vascular endothelial cells. In addition, through bioinformatics analysis using GEO data, changes in TOB2 and MEF2 were revealed in cardiovascular disease. We suggest that circulating miR-4701-3p has high potential as a biomarker for myocardial infarction.
血管生成主要受 VEGF 依赖性信号传递至细胞的调节。然而，VEGF 诱导的 miRNA 调节的血管生成机制仍不清楚。在 HUVECs 中经 VEGF 处理后，我们通过小 RNA 测序筛选出变化的 miRNA，发现 VEGF 诱导的 miR-4701-3p。此外，使用 GFP 报告基因揭示 TOB2 表达受 miR-4701-3p 调节，并且发现 TOB2 和 miR-4701-3p 调节可在体外血管生成测定中引起血管生成。通过荧光素酶测定，证实血管生成转录因子 MEF2 的激活受 TOB2 和 miR-4701-3p 的抑制和过表达调节。结果，调节了诱导血管生成功能的 MEF2 下游基因 mRNAs。我们使用 NCBI GEO 数据集揭示了心血管疾病中 TOB2 和 MEF2 的表达明显改变。最后，证实了心肌梗死患者血液中循环 miR-4701-3p 的表达显著增加。在心肌梗死患者中，循环 miR-4701-3p 增加，无论年龄、BMI 和性别如何，特异性和敏感性分析（AUROC）显示出高 AUC 水平（AUC=0.8451，95%CI 0.78-0.90）。我们的数据表明，VEGF 诱导的 miR-4701-3p 通过血管内皮细胞中的 TOB2 介导调节 MEF2 及其血管生成。此外，通过使用 GEO 数据进行生物信息学分析，揭示了心血管疾病中 TOB2 和 MEF2 的变化。我们建议循环 miR-4701-3p 作为心肌梗死的生物标志物具有很高的潜力。
