Ansary Stem Cell Institute, Division of Regenerative Medicine, Department of Medicine, Weill Cornell Medicine, New York, NY 10065.
Ronald O. Perelman and Claudia Cohen Center for Reproductive Medicine, Weill Cornell Medicine, New York, NY 10065.
Proc Natl Acad Sci U S A. 2021 Feb 23;118(8). doi: 10.1073/pnas.2016950118.
Cells derived from pluripotent sources in vitro must resemble those found in vivo as closely as possible at both transcriptional and functional levels in order to be a useful tool for studying diseases and developing therapeutics. Recently, differentiation of human pluripotent stem cells (hPSCs) into brain microvascular endothelial cells (ECs) with blood-brain barrier (BBB)-like properties has been reported. These cells have since been used as a robust in vitro BBB model for drug delivery and mechanistic understanding of neurological diseases. However, the precise cellular identity of these induced brain microvascular endothelial cells (iBMECs) has not been well described. Employing a comprehensive transcriptomic metaanalysis of previously published hPSC-derived cells validated by physiological assays, we demonstrate that iBMECs lack functional attributes of ECs since they are deficient in vascular lineage genes while expressing clusters of genes related to the neuroectodermal epithelial lineage (Epi-iBMEC). Overexpression of key endothelial ETS transcription factors (, , and ) reprograms Epi-iBMECs into authentic endothelial cells that are congruent with bona fide endothelium at both transcriptomic as well as some functional levels. This approach could eventually be used to develop a robust human BBB model in vitro that resembles the human brain EC in vivo for functional studies and drug discovery.
为了使体外多能来源的细胞在转录和功能水平上尽可能地类似于体内细胞，成为研究疾病和开发治疗方法的有用工具，这些细胞必须具有接近体内细胞的特性。最近，已有报道称可将人类多能干细胞（hPSC）分化为具有血脑屏障（BBB）特性的脑微血管内皮细胞（EC）。这些细胞已被用作药物输送和神经疾病机制理解的强大体外 BBB 模型。然而，这些诱导的脑微血管内皮细胞（iBMEC）的确切细胞特性尚未得到很好的描述。通过对以前发表的 hPSC 衍生细胞进行全面的转录组元分析，并通过生理测定进行验证，我们证明 iBMEC 缺乏 EC 的功能属性，因为它们缺乏血管谱系基因，而表达与神经外胚层上皮谱系（Epi-iBMEC）相关的基因簇。关键内皮 ETS 转录因子（、和）的过表达将 Epi-iBMEC 重编程为真正的内皮细胞，在转录组和一些功能水平上与真正的内皮细胞一致。这种方法最终可用于开发一种体外的稳健人类 BBB 模型，该模型在功能研究和药物发现方面类似于体内的人脑 EC。
