Target Assessment in Alzheimer's Neurovasculature/BBB

阿尔茨海默病神经血管系统/BBB 的目标评估

• 批准号: 6787895
• 负责人: JAN F SALLSTROM
• 金额: $ 10.7万
• 依托单位: SOCRATECH, LLC
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6787895
• 关键词: Alzheimer' s disease; astrocytes; bioassay; bioengineering /biomedical engineering; bioinformatics; biotechnology; blood brain barrier; cellular pathology; cerebrovascular system; disease /disorder etiology; functional /structural genomics; gene expression profiling; human tissue; immunologic techniques; laser capture microdissection; macrophage; microarray technology; molecular pathology; pathologic process; polymerase chain reaction; postmortem; proteomics; technology /technique development; tissue /cell culture; vascular endothelium

DESCRIPTION (provided by applicant): Socratech L.L.C. is a start-up company within the University of Rochester founded in July 2000 with a focus on vascular strategies for neurodegenerative disorders and Alzheimer's disease (AD). The goal of Phase I is to identify new therapeutic and diagnostic targets in AD neurovasculature/blood-brain barrier (BBB). Neurovasculature/BBB is comprised of brain endothelial cells (BEC), pericytes and astrocytes. Our central hypothesis is that AD could be primarily a neurovascular disorder and that dysfunctions in BEC, astrocytes and pericytes contribute significantly to the disease process. To optimize target assessment, we have developed a Process Biology approach which integrates several genomics and bioinformatics tools through different modules: the BioBank Socratech, BioAssay, Biolnfo, Microarray-Based Expression Profiling (MEP), Single-Target Expression Profiling (STEP), Histoproteomics and Rapid Inhibition of mRNA Expression (RIME). The BioBank Socratech with its 300 primary lines of BEC, pericytes, and glial cells from brains of AD patients and controls, tissue samples, RNA/DNA samples, RT-PCR primers and antibodies, is a central resource for this proposal. Our preliminary MEP/functional analysis of BEC and astrocytes supports the hypothesis that AD neurovascular cells are functionally and molecularly inherently different from controls and that they retain in culture certain pathoqenic molecular and cellular features observed in AD neurovasculature/BBB in brain tissue in situ. Phase I - Target Idenitification, will validate MEP/Functional data for selected Genes-of-Interest in AD and Controls through the STEP module by quantitative PCR in serial samples of cultured BEC and astrocytes, and in BEC and astrocytes isolated from brain tissue by Laser Capture Microscopy (aim 1); through the Histoproteomics module by immunodetection in serial brain tissues samples (aim 2); and will identify new Genes-of-Interest in pericytes by MEP (aim 3). Phase II will use STEP/RIME approach for Target Pathway Analysis and screening of different Libraries of Compounds, and Vectors for over-expression studies with drug-targetable genes. The Novelty is focus on the BBB in AD, and use of new in vitro neurovascular models in combination with LCM/QPCR tissue analysis to identify new targets. The potential products include new diagnostics DNA chips and brain imaging agents, therapeutic genetic vectors and new drugs to treat neurovascular AD disorder.

描述（由申请人提供）：Socratech L.L.C.是罗切斯特大学（University of Rochester）的一家初创公司，成立于2000年7月，着重于神经退行性疾病的血管策略和阿尔茨海默氏病（AD）。第一阶段的目的是确定AD神经血管形/血脑屏障（BBB）中的新治疗和诊断靶标。神经腔/BBB由脑内皮细胞（BEC），周细胞和星形胶质细胞组成。我们的中心假设是AD可能主要是一种神经血管疾病，并且BEC，星形胶质细胞和周细胞的功能障碍对疾病过程产生了重大贡献。为了优化目标评估，我们开发了一种过程生物学方法，该方法通过不同的模块来整合几种基因组学和生物信息学工具：Biobankank Socratech，Socratech，Bioassay，Biolnfo，基于微阵列的表达谱（MEP），单目标表达分析（STEP），组织蛋白质组学和快速抑制mRNA（RIME）（RIME）。来自AD患者和对照组，组织样本，RNA/DNA样品，RT-PCR引物和抗体的Biobank socratech及其BEC，周细胞和神经胶质细胞的30​​0条主要线条是该建议的核心资源。我们对BEC和星形胶质细胞的初步MEP/功能分析支持了以下假设：AD神经血管细胞在功能和分子上与对照固有不同，并且它们保留在某些病原体分子和细胞特征中在AD神经血管造影/BBB中观察到的某些病原体分子和细胞特征。第一阶段 - 目标iDenitifific将验证 AD中选定的利益基因的MEP/功能数据通过步骤模块通过 在培养的BEC和星形胶质细胞的串行样品以及通过激光捕获显微镜从脑组织中分离出的BEC和星形胶质细胞中的定量PCR（AIM 1）；通过串行脑组织样品中的免疫检测通过组织蛋白质组学模块（AIM 2）；并将通过MEP确定周细胞中的新基因（AIM 3）。 第二阶段将使用步骤/rime方法进行目标途径分析和筛选不同库 用药物靶向基因进行过表达研究的化合物和载体的载体。新颖性是专注于AD中的BBB，并将新的体外神经血管模型与LCM/QPCR组织分析结合使用以识别新靶标。潜在的产品包括新的诊断DNA芯片和脑成像剂，治疗遗传载体以及治疗神经血管AD疾病的新药物。