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: 6045298
• 负责人: PAULA GRAMMAS
• 金额: $ 32.31万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 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 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6045298
• 关键词: Alzheimer's disease; 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.

阿尔茨海默病 (AD) 是一种影响 400 万美国老年人的神经退行性疾病。 我们的假设是，脑微血管通过产生损伤或杀死神经元的可溶性因子而促成 AD 的发病机制。 我们小组的最新数据有力地支持了这一假设，并证明从 AD 患者中分离出的脑微血管和在培养物中受伤的大鼠脑内皮细胞都会分泌对神经元造成致命损伤的因子。我们已经产生了可识别并免疫沉淀大鼠内皮细胞衍生毒素的单克隆抗体。初步实验表明，这些抗体中至少有一种也能免疫沉淀人AD微血管分泌的神经毒性因子，表明内皮和微血管神经毒素是相同或密切相关的。 这些新发现令人兴奋，并暗示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有力地支持了实现该项目目标的可行性，即纯化和鉴定这些血管源性神经毒性因子。 在这个项目中，我们的具体目标是：1）纯化和表征大鼠内皮细胞源性毒性因子（EDTF）； 2）克隆EDTF的cDNA并确定杀伤神经元的结构基础； 3) 表征针对人 AD 微血管源性神经毒性因子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。 我们的实验首次确定了血管系统是 AD 中神经毒性分子的来源。 无论神经毒性因子是已知的还是独特的，正如本项目中将评估的那样，AD脑微血管产生损伤或杀死神经元的可溶性因子的证明表明，大脑微循环是 AD 中神经毒性因子的一种新的、未经探索的来源，并且迄今为止尚未认识到的治疗干预目标。