From the Hans Schöler Stem Cell Research Center, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), South Korea (S.Y.P., H.L., M.R.P., J.B.K.).
Department of Physiology, Pusan National University School of Medicine, Yangsan, South Korea (Y.W.K., J.H.K.).
Arterioscler Thromb Vasc Biol. 2020 Apr;40(4):e105-e113. doi: 10.1161/ATVBAHA.119.313684. Epub 2020 Feb 20.
Vascular progenitor cells (VPCs), which are able to differentiate into both endothelial cells and smooth muscle cells, have the potential for treatment of ischemic diseases. Generated by pluripotent stem cells, VPCs carry the risk of tumorigenicity in clinical application. This issue could be resolved by direct lineage conversion, the induction of functional cells from another lineage by using only lineage-restricted transcription factors. Here, we show that induced VPCs (iVPCs) can be generated from fibroblasts by ETS (E-twenty six) transcription factors, and . Approach and Results: Mouse fibroblasts were infected with lentivirus encoding and . Cell colonies appeared in - and -infected groups and were mechanically picked. The identity of cell colonies was confirmed by proliferation assay and reverse-transcription polymerase chain reaction with vascular markers- infected cell colonies were sorted by CD144 (also known as CDH5, VE-cadherin). We defined that CD144-positive iVPCs maintained its own population and expanded stably at multiple passages. iVPCs could differentiate into functional endothelial cells and smooth muscle cells by a defined medium. The functionalities of iVPC-derived endothelial cells and smooth muscle cells were confirmed by analyzing LDL (low-density lipoprotein) uptake, carbachol-induced contraction, and tube formation in vitro. Transplantation of iVPCs into the ischemic hindlimb model enhanced blood flow without tumor formation in vivo. Human iVPCs were generated by human ETS transcription factors and .
We demonstrate that ischemic disease curable iVPCs, which have self-renewal and bipotency, can be generated from mouse fibroblasts by enforced ETS family transcription factors, and expression. Our simple strategy opens insights into stem cell-based ischemic disease therapy.
血管祖细胞（VPCs）能够分化为内皮细胞和平滑肌细胞，具有治疗缺血性疾病的潜力。由多能干细胞产生的 VPCs 在临床应用中存在致瘤性风险。通过直接谱系转换，可以解决这个问题，即用仅受谱系限制的转录因子从另一个谱系诱导功能性细胞。在这里，我们展示了可以通过 ETS（E-twenty six）转录因子和 从成纤维细胞中诱导产生 VPCs（iVPCs）。
用编码 和 的慢病毒感染小鼠成纤维细胞。在 和 感染组中出现细胞集落，并通过机械方法挑取。通过增殖试验和血管标记物的逆转录聚合酶链反应（vascular markers-）证实细胞集落的身份-感染的细胞集落通过 CD144（也称为 CDH5、VE-钙粘蛋白）进行分选。我们定义 CD144 阳性 iVPCs 保持自身群体，并在多个传代中稳定扩增。iVPCs 可以在定义的培养基中分化为功能性内皮细胞和平滑肌细胞。通过分析 LDL（低密度脂蛋白）摄取、卡巴胆碱诱导的收缩和体外管状形成，证实了 iVPC 衍生的内皮细胞和平滑肌细胞的功能。将 iVPCs 移植到缺血性后肢模型中，可在体内增强血液流动而无肿瘤形成。用人 ETS 转录因子 和 生成人 iVPCs。
我们证明了可治愈缺血性疾病的 iVPCs 具有自我更新和双潜能，可以通过强制表达 ETS 家族转录因子 和 从小鼠成纤维细胞中产生。我们的简单策略为基于干细胞的缺血性疾病治疗提供了新的思路。
