PDE3A通过内皮细胞调控血脑屏障功能参与脑小血管病发生发展的机制研究

内皮细胞是血脑屏障的关键组成部分，与周围细胞互作以维持血管稳态。内皮损伤是脑小血管病的起始事件，可引发血管内炎症和血脑屏障破坏，进而导致相关脑损害。本团队前期从中国国家卒中登记III研究中发现，PDE3A基因与脑小血管病相关；研究表明该基因参与调节内皮屏障功能与维持血管完整性。由此我们推测：PDE3A变化可引起内皮细胞激活和损伤，从而导致血脑屏障破坏及脑小血管病相关病理改变。而PDE3A参与内皮损伤及脑小血管病发生发展的机制有待深入研究。本项目拟采用CRISPR-Cas9技术分别构建PDE3A功能缺失和增强的模型鼠；在不同时间点（鼠龄第4-72周），评估内皮及脑小血管病相关表型及病理变化，评估血管炎症、血脑屏障完整性、胶质细胞变化及内皮细胞中基因表达差异，发掘相关分子网络，揭示PDE3A通过内皮细胞调控血脑屏障功能参与脑小血管病发生发展机制，为寻找脑小血管病新治疗靶点提供科学依据。

Endothelial cells are key components of the blood-brain barrier and interact with surrounding cells to maintain vascular homeostasis. Endothelial injury is the initial event of cerebral small vessel disease, which can lead to intravascular inflammation, destruction of the blood-brain barrier, and related brain damage. We have previously found that PDE3A gene was associated with cerebral small vessel disease from the China National Stroke Registry III (CNSR-III). PDE3A gene plays significant roles in regulation of endothelial function and maintenance of vascular integrity. Thus, we hypothesize that alteration of PDE3A can cause activation and injury of endothelial cells, which will lead to damage of blood-brain barrier and pathological changes related to cerebral small vessel disease. Nevertheless, the mechanism of PDE3A leading to endothelial dysfunction and cerebral small vessel disease remains to be further investigated. In this study, we intend to use CRISPR-Cas9 tool to generate mouse models of PDE3A function deletion or enhancement, respectively. Endothelial function, phenotypes and pathological changes related to cerebral small vessel disease will be assessed, at different time points (4-72 weeks of age). The differences of vascular inflammation, blood-brain barrier integrity, glial cells and gene expression in endothelial cells among different mouse models will also be evaluated. These researches will help to reveal the mechanism of PDE3A promoting the occurrence and development of cerebral small vessel disease through endothelial injury, and provide a substantial basis for searching for new therapeutic targets.