Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
Departments of Neurosciences and Pharmacology, University of California, San Diego, La Jolla, CA 92093, USA.
Sci Adv. 2024 Feb 2;10(5):eadi1737. doi: 10.1126/sciadv.adi1737.
Brain mural cells regulate development and function of the blood-brain barrier and control blood flow. Existing in vitro models of human brain mural cells have low expression of key mural cell genes, including . Thus, we asked whether activation of Notch3 signaling in hPSC-derived neural crest could direct the differentiation of brain mural cells with an improved transcriptional profile. Overexpression of the Notch3 intracellular domain (N3ICD) induced expression of mural cell markers PDGFRβ, TBX2, , , , and endogenous Notch3. The resulting N3ICD-derived brain mural cells produced extracellular matrix, self-assembled with endothelial cells, and had functional K channels. ChIP-seq revealed that Notch3 serves as a direct input to relatively few genes in the context of this differentiation process. Our work demonstrates that activation of Notch3 signaling is sufficient to direct the differentiation of neural crest to mural cells and establishes a developmentally relevant differentiation protocol.
脑壁细胞调节血脑屏障的发育和功能，并控制血流。现有的人脑壁细胞体外模型中，关键的壁细胞基因表达水平较低，包括. 因此，我们想知道 Notch3 信号在 hPSC 衍生的神经嵴中的激活是否可以指导具有改善的转录谱的脑壁细胞的分化。Notch3 细胞内结构域（N3ICD）的过表达诱导了壁细胞标志物 PDGFRβ、TBX2、、、和内源性 Notch3 的表达。由此产生的 N3ICD 衍生的脑壁细胞产生细胞外基质，与内皮细胞自我组装，并具有功能性 K 通道。ChIP-seq 表明，Notch3 在这种分化过程中作为相对较少基因的直接输入。我们的工作表明，Notch3 信号的激活足以指导神经嵴向壁细胞的分化，并建立了一个具有发育相关性的分化方案。
