VASCULAR-MEDIATED NEURONAL CELL DEATH IN ALZHEIMER'S

阿尔茨海默病中血管介导的神经细胞死亡

• 批准号: 6988276
• 负责人: PAULA GRAMMAS
• 金额: $ 2.87万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6988276
• 关键词: Alzheimer's disease; brain circulation; cell death; clinical research; human tissue; immunocytochemistry; laboratory mouse; laboratory rat; microcirculation; molecular cloning; monoclonal antibody; neurons; neurotoxicology; neurotoxins; postmortem; protein localization; protein purification; protein structure function; tissue /cell culture; vascular endothelium

Alzheimer's disease (AD) is a neurodegenerative disorder affecting 4 million older Americans. Our hypothesis is that brain microvessels contribute to the pathogenesis of AD by producing soluble factors that injure or kill neurons. Recent data from our group strongly support this hypothesis and demonstrate that both brain microvessels isolated from AD patients and rat brain endothelial cells injured in culture secrete factors that cause lethal injury specifically to neurons. We have generated monoclonal antibodies that recognize and immunoprecipitate the rat endothelial cell-derived toxin. Preliminary experiments show that at least one of these antibodies also immunoprecipitates the neurotoxic factor secreted by human AD microvessels, suggesting that the endothelial and microvessel neurotoxins are identical or closely related. These novel findings are exciting and implicate the vasculature as a source of neurotoxins in the brain. The feasibility of achieving the goals of this project, to purify and identify these vascular- derived neurotoxic factors, is strongly supported by our initial data and the availability of monoclonal antibodies. In this project, our specific aims are: 1) to purify and characterize the rat endothelial cell-derived toxic factor (EDTF); 2) to clone the cDNA for EDTF and determine the structural basis for neuronal killing; 3) to characterize multi-functional monoclonal antibodies to the human AD microvessel-derived neurotoxic factor(s); and 4) to determine the localization of neurotoxin(s) in normal and AD tissues. Our experiments are the first to identify the vasculature as a source of neurotoxic molecules in AD. Whether the neurotoxic factor is known or unique, as will be assessed in this project, the demonstration that AD brain microvessels produce soluble factors that injure or kill neurons shows that the cerebral microcirculation is a novel, unexplored source of neurotoxic factors in AD, and a heretofore unrecognized target for therapeutic intervention.

阿尔茨海默氏病（AD）是一种影响400万美国人的神经退行性疾病。 我们的假设是，脑微血管通过产生损害或杀死神经元的可溶性因子来促进AD的发病机理。 来自我们小组的最新数据强烈支持这一假设，并表明，在培养物中受伤的AD患者和大鼠脑内皮细胞分离出的脑微血管都会分泌专门针对神经元致命损伤的因素。我们已经产生了识别和免疫沉淀大鼠内皮细胞衍生的毒素的单克隆抗体。初步实验表明，这些抗体中的至少一种还可以免疫沉淀人类AD微血管分泌的神经毒性因子，这表明内皮和微血管神经毒素是相同或密切相关的。 这些新颖的发现令人兴奋，并暗示了脉管系统是大脑中神经毒素的来源。 我们的初始数据和单克隆抗体的可用性强烈支持实现该项目的目标，净化和识别这些血管衍生的神经毒性因子的可行性。 在这个项目中，我们的具体目的是：1）净化和表征大鼠内皮细胞衍生的毒性因子（EDTF）； 2）克隆EDTF的cDNA并确定神经元杀死的结构基础； 3）表征与人AD微血管衍生的神经毒性因子（S）的多功能单克隆抗体； 4）确定神经毒素在正常组织和AD组织中的定位。 我们的实验是第一个将脉管系统鉴定为AD中神经毒性分子的来源的实验。 在该项目中将评估神经毒性因子是已知还是独特的，这证明了AD脑微血管产生可溶性因子，损害或杀死神经元的可溶性因子表明，大脑微循环是一种新型的，未探索的AD和A AD型神经毒性因子的来源，以及A次托未识别的治疗靶标。