MicroRNA-199b Modulates Vascular Cell Fate During iPS Cell Differentiation by Targeting the Notch Ligand Jagged1 and Enhancing VEGF Signaling.
MicroRNA-199b Modulates Vascular Cell Fate During iPS Cell Differentiation by Targeting the Notch Ligand Jagged1 and Enhancing VEGF Signaling.
复制标题
MicroRNA-199b 通过靶向 Notch 配体 Jagged1 并增强 VEGF 信号传导来调节 iPS 细胞分化过程中的血管细胞命运
DOI：
10.1002/stem.1930
复制
发表时间：
2015-05
期刊：
影响因子：
--
通讯作者：
Xu Q
中科院分区：
文献类型：
--
作者：
Chen T;Margariti A;Kelaini S;Cochrane A;Guha ST;Hu Y;Stitt AW;Zhang L;Xu Q
目的：近年来，从诱导多能干细胞(IPS)中诱导内皮细胞(ECs)的能力为个体化药物和干细胞治疗提供了巨大的治疗潜力。我们的目标是，更好地了解iPS细胞向内皮细胞分化过程中诱发的复杂分子信号，可能会允许特定的靶向其活性，以促进细胞分化和促进组织再生。方法和结果：在这项研究中，我们使用既定的方案从成纤维细胞中产生了小鼠iPS细胞。将iPS细胞接种于小鼠IV型胶原包被的培养皿中，观察到iPS细胞向血管方向分化。为了研究iPS细胞分化的分子机制，我们发现miR-199B参与了EC的分化。在EC分化过程中，miR-199的表达呈递增趋势。值得注意的是，miR-199B靶向Notch配体JAG1，导致血管内皮生长因子(VEGF)通过转录因子STAT3转录激活和分泌。在shRNA介导的Notch配体JAG1被敲除后，miR-199B的调节作用被消融，在EC分化过程中STAT3和VEGF被强烈诱导。JAG1基因敲除也可抑制miR-199B介导的iPS细胞向平滑肌标记物分化的抑制。通过体外管形成实验和植入Matrigel塞子，在体内，miR-199B还调节了血管内皮生长因子的表达和血管生成。结论：本研究表明miR-199B通过调节关键信号血管生成反应，在iPS细胞分化的血管细胞过程中作为表型转换的调节因子发挥了新的作用。干细胞2015；33：1405-1418
Aims: Recent ability to derive endothelial cells (ECs) from induced pluripotent stem (iPS) cells holds a great therapeutic potential for personalized medicine and stem cell therapy. We aimed that better understanding of the complex molecular signals that are evoked during iPS cell differentiation toward ECs may allow specific targeting of their activities to enhance cell differentiation and promote tissue regeneration. Methods and Results: In this study, we have generated mouse iPS cells from fibroblasts using established protocol. When iPS cells were cultivated on type IV mouse collagen‐coated dishes in differentiation medium, cell differentiation toward vascular lineages were observed. To study the molecular mechanisms of iPS cell differentiation, we found that miR‐199b is involved in EC differentiation. A step‐wise increase in expression of miR‐199 was detected during EC differentiation. Notably, miR‐199b targeted the Notch ligand JAG1, resulting in vascular endothelial growth factor (VEGF) transcriptional activation and secretion through the transcription factor STAT3. Upon shRNA‐mediated knockdown of the Notch ligand JAG1, the regulatory effect of miR‐199b was ablated and there was robust induction of STAT3 and VEGF during EC differentiation. Knockdown of JAG1 also inhibited miR‐199b‐mediated inhibition of iPS cell differentiation toward smooth muscle markers. Using the in vitro tube formation assay and implanted Matrigel plugs, in vivo, miR‐199b also regulated VEGF expression and angiogenesis. Conclusions: This study indicates a novel role for miR‐199b as a regulator of the phenotypic switch during vascular cell differentiation derived from iPS cells by regulating critical signaling angiogenic responses. Stem Cells 2015;33:1405–1418