Direct Reprogramming of Human Dermal Fibroblasts Into Endothelial Cells Using ER71/ETV2.
Direct Reprogramming of Human Dermal Fibroblasts Into Endothelial Cells Using ER71/ETV2.
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DOI：
10.1161/circresaha.116.309833
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发表时间：
2017-03-03
影响因子：
20.1
通讯作者：
Yoon YS
中科院分区：
文献类型：
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作者：
Lee S;Park C;Han JW;Kim JY;Cho K;Kim EJ;Kim S;Lee SJ;Oh SY;Tanaka Y;Park IH;An HJ;Shin CM;Sharma S;Yoon YS
绕过干细胞或祖细胞状态，将人类出生后细胞直接转化或重编程为内皮细胞（EC）对于再生医学、细胞治疗和病理生理学研究至关重要，但在很大程度上仍未得到探索。我们试图将人出生后真皮成纤维细胞（HDF）直接重编程为具有血管生成和内皮转录因子（TF）的EC，并确定其血管化和治疗潜力。我们利用七个 EC TF 的各种组合来转导 HDF，并发现单独的 ER71/ETV2 可以最好地诱导内皮特征。第 7 天从 ER71/ETV2 转导的 HDF 中分选的 KDR+ 细胞显示出不太成熟但丰富的内皮特征，因此被称为早期重编程 EC (rEC)，并且不会通过进一步培养而成熟。经过几周的无转基因培养，然后短暂重新诱导 ER71/ETV2，早期 rEC 在三个月的培养过程中成熟，并显示 ETV2 表达减少，达到与出生后人类 EC 相似的成熟表型。这些被称为晚期 rEC。虽然早期 rEC 表现出不成熟的表型，但将它们植入缺血后肢可促进缺血恢复。这两种 rEC 显示出通过体内直接血管合并促进新血管形成的明显能力。植入的早期 rEC 的旁分泌或促血管生成作用在修复后肢缺血方面发挥着重要作用。这项研究首次证明，在无转基因干预期后，单独的 ER71/ETV2 可以直接将人类出生后细胞重编程为有功能的成熟 EC。这些 rEC 对于细胞治疗、个性化疾病研究和重编程过程的探索可能很有价值。
Direct conversion or reprogramming of human postnatal cells into endothelial cells (ECs), bypassing stem or progenitor cell status, is crucial for regenerative medicine, cell therapy, and pathophysiological investigation but has remained largely unexplored. We sought to directly reprogram human postnatal dermal fibroblasts (HDFs) to ECs with vasculogenic and endothelial transcription factors (TFs) and determine their vascularizing and therapeutic potential. We utilized various combinations of seven EC TFs to transduce HDFs and found that ER71/ETV2 alone best induced endothelial features. KDR+ cells sorted at day 7 from ER71/ETV2-transduced HDFs showed less mature but enriched endothelial characteristics and thus were referred to as early reprogrammed ECs (rECs), and did not undergo maturation by further culture. After a period of several weeks’ transgene-free culture followed by transient re-induction of ER71/ETV2, early rECs matured during three months of culture and showed reduced ETV2 expression, reaching a mature phenotype similar to postnatal human ECs. These were termed late rECs. While early rECs exhibited an immature phenotype, their implantation into ischemic hindlimbs induced enhanced recovery from ischemia. These two rECs showed clear capacity for contributing to new vessel formation through direct vascular incorporation in vivo. Paracrine or pro-angiogenic effects of implanted early rECs played a significant role in repairing hindlimb ischemia. This study for the first time demonstrates that ER71/ETV2 alone can directly reprogram human postnatal cells to functional, mature ECs after an intervening transgene free period. These rECs could be valuable for cell therapy, personalized disease investigation, and exploration of the reprogramming process.