脑血管内皮细胞起源和发育及其调控机制

Cerebral vessel abnormalities are the major causes of morbidity and mortality worldwide, but the underlying molecular pathology remains incompletely understood and clinical interventions are very limited. By applying zebrafish embryology and genetics, high-resolution confocal imaging, single-cell RNA-seq, ChIP-seq and others, we plan to 1) investigate the origin and development of cerebral vascular endothelial cells, as well as its potential regulation by transcriptional factor Npas4l and others; 2) study the role of WNT signaling (ligands and receptors) in regulating the differentiation of anterior lateral plate mesoderm into cerebral endothelial cells; and 3) explore the function of betaPix and interacting proteins in regulating maturation of cerebral vessels. This proposal is highly supported by our previous observations that the dorsal, but not ventral, lateral cells of shield embryos contribute to the formation of cerebral endothelial cells, which requires down-regulation of the canonical WNT signaling, and betaPix is essential for stabilization of cerebral vessels. Taken together, our studies may allow us to discover new principles in the field of cerebral vessel development, as well as improve differentiation of human embryonic stem cells or direct transdifferentiation of fibroblasts into endothelial cells for cerebral vessel repair and regeneration.

脑血管疾病是严重影响人类生命和健康的重大疾病，目前针对脑血管病缺乏理想的治疗药物。本项目利用斑马鱼胚胎学和遗传学手段，结合大视场高分辨显微成像、单细胞转录组分析、ChIP-Seq等技术，深入研究斑马鱼脑血管内皮细胞的起源，以及Npas4l等调控中胚层细胞特化成为内皮祖细胞的分子机制；探讨调控从前侧板中胚层发育成脑血管的信号通路，尤其集中研究WNT信号调控脑血管发育的分子机制；研究betaPix及相关因子在脑血管成熟和稳态维持中的功能。我们前期工作发现囊胚期背侧面细胞参与前侧板中胚层和脑血管内皮细胞发育，并且此发育过程受到WNT信号通路负调控；betaPix突变体中MMP9过表达导致脑血管破裂出血，这些数据为本项目提供了扎实的基础。本研究将不仅完善脑血管发育生物学理论，而且也为指导人多能干细胞定向分化或转分化成为血管内皮细胞以及改善脑血管损伤修复提供理论依据。

脑血管疾病是严重影响人类生命和健康的重大疾病，目前针对脑血管病缺乏理想的治疗药物。本项目利用斑马鱼胚胎学和遗传学手段，结合大视场高分辨显微成像等技术，深入研究了斑马鱼脑血管内皮细胞的起源，发现胚盾期背侧区代表头部血管内皮祖细胞特化区域。从单细胞转录组分析、ChIP-Seq等技术，探讨调控从前侧板中胚层发育成脑血管的信号通路，发现WNT信号负调控脑血管发育的机制。建立了betaPix条件性敲除和敲入斑马鱼品系，发现betaPix基因富集于中后脑交界区，神经胶质细胞特异性敲除betaPix导致脑出血，提示胶质细胞betaPix在调节脑出血中的重要功能。通过比较betaPix突变体与对照胚胎单细胞转录组，将有利于揭示神经胶质细胞在脑血管发育与稳态中的功能。本研究不仅完善脑血管发育生物学的理论，而且也为指导人多能干细胞定向分化或转分化成为血管内皮细胞以及改善脑血管损伤修复提供理论依据。