Conversion of human fibroblasts to functional endothelial cells by defined factors.
Conversion of human fibroblasts to functional endothelial cells by defined factors.
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DOI：
10.1161/atvbaha.112.301167
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发表时间：
2013-06
期刊：
影响因子：
--
通讯作者：
Ding S
中科院分区：
文献类型：
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作者：
Li J;Huang NF;Zou J;Laurent TJ;Lee JC;Okogbaa J;Cooke JP;Ding S
成纤维细胞向内皮细胞（EC）的转分化可能为缺血和纤维化等疾病提供新的治疗途径。在这里，我们证明，在诱导信号条件下，仅使用 Oct4 和 Klf4 两个因子即可将人成纤维细胞转分化为功能性 EC。为了确定人成纤维细胞是否可以通过多能因子的瞬时表达转化为EC，在存在促进内皮细胞程序诱导的可溶性因子的情况下，用编码Oct4和Klf4的慢病毒转导人新生儿成纤维细胞。 28 天后，诱导内皮 (iEnd) 细胞簇出现，并被分离以进行进一步增殖和后续表征。 iEnd 细胞通过表达 CD31、VE-钙粘蛋白和血管性血友病因子等内皮表型标记物，在转录特征上与原代人类 EC 相似。此外，iEnd 细胞可以掺入乙酰化低密度脂蛋白，并在体外和体内形成血管结构。当注射到小鼠缺血肢体时，iEnd 细胞被移植，毛细血管密度增加，组织灌注增强。在转分化过程中，内源多能网络没有激活，表明该过程绕过了多能中间步骤。多能因子诱导的转分化可成功用于生成用于治疗应用的功能性自体 EC。
Transdifferentiation of fibroblasts to endothelial cells (ECs) may provide a novel therapeutic avenue for diseases including ischemia and fibrosis. Here we demonstrate that human fibroblasts can be transdifferentiated into functional ECs by using only two factors, Oct4 and Klf4, under inductive signaling conditions. To determine if human fibroblasts could be converted into ECs by transient expression of pluripotency factors, human neonatal fibroblasts were transduced with lentiviruses encoding Oct4 and Klf4 in the presence of soluble factors that promote the induction of an endothelial program. After 28 days, clusters of induced endothelial (iEnd) cells appeared and were isolated for further propagation and subsequent characterization. The iEnd cells resembled primary human ECs in their transcriptional signature by expressing endothelial phenotypic markers such as CD31, VE-cadherin, and von Willebrand Factor. Furthermore, the iEnd cells could incorporate acetylated low density lipoprotein, and form vascular structures in vitro and in vivo. When injected into the ischemic limb of mice, the iEnd cells engrafted, increased capillary density, and enhanced tissue perfusion. During the transdifferentiation process, the endogenous pluripotency network was not activated, suggesting that this process bypassed a pluripotent intermediate step. Pluripotent factor–induced transdifferentiation can be successfully applied for generating functional autologous ECs for therapeutic applications.