靶向激活脑血管内皮SIRT1用于治疗肾上腺脑白质萎缩症的探究

项目名称： 靶向激活脑血管内皮SIRT1用于治疗肾上腺脑白质萎缩症的探究

项目编号： No.81501038

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 龚昳

作者单位： 徐州医科大学

项目金额： 17.5万元

中文摘要： X-ALD的进程与血脑屏障结构和功能的改变密切相关，但其机制不明。我们前期的工作证实：X-ALD的致病基因ABCD1的 沉默引发大脑血管内皮细胞特征性蛋白分子的表达和定位显著改变，致使内皮细胞功能严重失衡，同时伴发SIRT1表达的显著下调。由此，我们提出以下假设：ABCD1基因缺失，通过影响SIRT1的表达及其功能进而导致血管内皮特征性蛋白分子发生变化；最终导致内皮细胞功能失衡，是X-ALD的主要病理生理机制。本课题首次采用ABCD1缺失的大脑血管内皮细胞特征性分子表达和功能变化这一模型，应用基因敲除、小干扰RNA、免疫印迹、流式细胞等技术，研究SIRT1在ABCD1缺失大脑血管内皮细胞中的作用，并探讨药物和转基因技术提升SIRT1水平对血管内皮功能的影响以及对X-ALD疾病进程的影响。本结果将首次揭示ABCD1基因突变导致血脑屏障破坏的分子机制，为X-ALD的靶向治疗提供理论依据。

中文关键词： X-染色体连锁肾上腺脑白质萎缩症；ATP；结合转运子超家族成员；D1；；脑血管内皮；SIRT1；单核/巨噬细胞

英文摘要： Disruption of blood-brain barrier plays key role in the development of X-ALD disease, yet the mechanism remains elusive. Our previous work evidenced that the silencing of X-ALD disease-causing gene ABCD1 induced remarkable alteration of protein markers (both expression and localization) as well as functional deterioration in brain microvascular endothelial cells. All these changes was accompanied by dramatic downregulation of SIRT1 . Herein, we propose that ABCD1 deficiency leads to protein marker expression changes through SIRT1, then further functional damage in brain microvascular cells. We therefore will use the established ABCD1 deficient brain microvascular endothelial cell system to test our hypothesis. We will first use RNAi, immunoblot and flow cytometry to determine the function of SIRT1 in ABCD1 deficiency caused endothelial cell dysfunction. Afterward, we will improve SIRT1 expression and function via drug treatment or transgenic technique to test SIRT1 improvement on molecular marker and functional enhancement in ABCD1-deficient brain endothelial cells, thereby further impact on disease progression. The proposed study will test a novel hypothesis using innovative and advanced technology. The outcome could help delving into the molecular mechanism of blood-brain barrier disruption resulting from ABCD1 mutation. The results would likely provide theoretical support for the development of targeted intervention of X-ALD.

英文关键词： X-linked adrenoleukodystrophy;ABCD1;brain microvascular endothelium;SIRT1;monocyte/macrophage