The Microvasculature in Alzheimer’s Disease: Viable Microvessels for Studies of Brain Endothelial and Glycocalyx Health

阿尔茨海默氏病的微血管系统：用于脑内皮和糖萼健康研究的活性微血管

• 批准号: 10352021
• 负责人: MAY J REED
• 金额: $ 15.55万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10352021
• 关键词: Alzheimer' s Disease; Autopsy; Biological Assay; Blood - brain barrier anatomy; Blood Circulation; Brain; Cell Aging; Cell Count; Cell Survival; Cell physiology; Clinical; Dementia; Endothelium; Enzymes; Extracellular Matrix; Extracellular Matrix Degradation; Foundations; Freezing; Gel; Glycocalyx; Glypican; HAS2 gene; Health; Heparan Sulfate Proteoglycan; Histologic; Hour; Hyaluronan; Impairment; Intervention; Knowledge; Label; Liquid substance; Measures; Metabolic; Methods; Movement; Outcome; Outcome Measure; Phenotype; Prevalence; Process; Proteins; Sampling; Specimen; Stains; Structure; Sulfate; Surface; Testing; Therapeutic; Thinness; Tissues; Transcript; base; beta-Galactosidase; biglycan; brain endothelial cell; brain parenchyma; brain tissue; cerebral microvasculature; clinical diagnosis; enzyme activity; experimental study; human subject; innovation; insight; neuropathology; neurovascular unit; proteoglycan core protein; senescence; syndecan; tissue culture

ABSTRACT/SUMMARY Brain microvasculature is altered in Alzheimer’s Disease (AD), but there are few methods to examine cellular processes in living tissue. Viable brain microvessels (MV) can be isolated from whole brain tissue of subjects undergoing rapid autopsy (<12 hours post mortem) in the UW Neuropathology (NP) Core. The donor subjects in the UW NP Core have a clinical diagnosis of dementia or no dementia, and all brain specimens undergo extensive neuropathologic assessment. This proposal utilizes innovative methods to rapidly obtain and maintain viable brain MV for a minimum of 5 days in supportive liquid media under standard tissue culture conditions, which affords a unique opportunity to study the microvasculature in brains with and without AD. Many aspects of MV structure and composition contribute to interactions that are relevant to AD. For this R03, the focus is on the luminal surface and the 2 components that are the initial interface between the brain and the systemic circulation: 1) the brain microvascular endothelial cell (BEC), and 2) the glycocalyx, a dynamic gel-like layer of extracellular matrix (ECM) that lines the luminal surface of BEC. The following 2 hypotheses will be tested. First, MV from those with dementia have similar BEC-specific viability and overall metabolic activity, but increased prevalence of senescent BEC relative to MV from those without dementia. Second, MV from those with dementia have greater glycocalyx shedding, which reflects increased degradative enzymes, relative to MV from those without dementia. Samples and outcome measures will be initially examined based on the clinical diagnosis of dementia or no dementia, and then re-classified according to subsequent detailed neuropathologic criteria. Aim 1 will define how BEC from dementia and no dementia MV specimens differ with respect to outcome measures of BEC-specific viability, overall metabolic activity, and BEC senescence. Aim 2 will determine how the glycocalyx from dementia and no dementia MV differ in de novo synthesis of ECM and markers of ECM degradation. Studies of glycocalyx will focus on heparan sulfate proteoglycans (HSPGs) and hyaluronan (HA). Knowledge gained will provide the foundation for perturbation/intervention experiments that are feasible, and have therapeutic implications for the mitigation of deleterious MV changes with dementia.

摘要/总结 阿尔茨海默病 (AD) 中大脑微血管系统发生改变，但几乎没有方法可以改变 检查活组织中的细胞过程。可以分离出活的脑微血管（MV） 在华盛顿大学接受快速尸检（死后 12 小时内）的受试者的全脑组织 神经病理学（NP）核心。 UW NP 核心的供体受试者具有以下临床诊断： 痴呆或无痴呆，所有脑标本均经过广泛的神经病理学检查 评估。该提案利用创新方法快速获得并维持存活的大脑 在标准组织培养条件下，在支持性液体培养基中 MV 至少 5 天， 这为研究患有和不患有 AD 的大脑微血管系统提供了独特的机会。 MV 结构和组成的许多方面都有助于与相关的相互作用 广告。对于此 R03，重点是管腔表面和初始的 2 个组件 大脑与体循环的界面：1）大脑微血管内皮 细胞 (BEC)，以及 2) 糖萼，细胞外基质 (ECM) 的动态凝胶状层， BEC 的管腔表面。 将检验以下 2 个假设。首先，痴呆症患者的 MV 具有相似的特征 BEC 特异性活力和总体代谢活性，但衰老 BEC 的患病率增加 相对于那些没有痴呆的人的 MV。其次，痴呆症患者的 MV 更大 糖萼脱落，反映了相对于 MV 而言，降解酶的增加 没有痴呆症。样本和结果测量将根据临床情况进行初步检查 诊断痴呆或无痴呆，然后根据后续详细信息重新分类 神经病理学标准。 目标 1 将定义痴呆 BEC 和非痴呆 MV 样本之间的差异 BEC 特异性活力、总体代谢活性和 BEC 衰老的结果测量。 目标 2 将确定痴呆症 MV 和非痴呆 MV 的糖萼在新过程中有何不同 ECM 的合成和 ECM 降解标记物。糖萼的研究将集中在乙酰肝素 硫酸蛋白聚糖（HSPG）和透明质酸（HA）。获得的知识将提供 为可行且具有治疗作用的扰动/干预实验奠定了基础 对减轻痴呆症引起的有害 MV 变化的影响。