Structural Identification and Functional Consequences of Different Amyloid Strains in Alzheimer's Disease

阿尔茨海默病中不同淀粉样蛋白菌株的结构鉴定和功能后果

• 批准号: 9789805
• 负责人: STEVEN Owen SMITH
• 金额: $ 62.66万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9789805
• 关键词: Academic Medical Centers; Address; Age; Age of Onset; Aging; Alzheimer' s Disease; Amyloid; Amyloid beta-Protein; Amyloid deposition; Apolipoprotein E; Atomic Force Microscopy; Biological Markers; Brain; Cell Culture Techniques; Cells; Characteristics; Clinical; Dementia; Deposition; Disease; Early Onset Familial Alzheimer' s Disease; Fluorescence Spectroscopy; Gender; Genetic Polymorphism; Genetic screening method; Genotype; Gossypium; Growth; Human; In Vitro; Inflammatory Response; Knowledge; Late Onset Alzheimer Disease; Location; Measurement; Microglia; Morphology; Mutation; NMR Spectroscopy; Netherlands; Onset of illness; Pathologic; Pathology; Peptides; Pharmaceutical Preparations; Presenile Alzheimer Dementia; Proteins; Rattus; Reporting; Sampling; Seeds; Senile Plaques; Spectroscopy, Fourier Transform Infrared; Spectrum Analysis; Structure; Symptoms; Tissue Sample; Toxic effect; Transmission Electron Microscopy; Wool; amyloid formation; beta pleated sheet; brain parenchyma; brain tissue; disorder subtype; familial Alzheimer disease; illness length; in vivo; microscopic imaging; neuroimaging; neuroinflammation; presenilin-2; solid state nuclear magnetic resonance; tau Proteins

Alzheimer's disease is associated with the deposition of amyloid in the brain during aging. Since the correlation between amyloid formation and AD was originally made, it has been recognized that there are many subtypes and forms that the disease can take. For example, accumulation of the amyloid β-protein (Aβ) in the brain parenchyma is the hallmark of Alzheimer's disease (AD). Nevertheless, there is a poor understanding as to why amyloid forms, and it is not known whether there are unique structural motifs that promote the distinct pathological consequences leading to dementia. The focus of this proposal is to fill this critical void in knowledge. Accordingly, the overall hypothesis of this proposal is that the Aβ peptides forming amyloid with distinct subtypes have distinct structures that determine their location and pathology. To address this hypothesis we propose two specific aims. First, we plan to isolate amyloid from different subtypes of post mortem brain tissue of late-onset AD and early- onset familial AD. We plan to compare five different amyloid plaque subtypes: typical AD, atypical AD, cotton wool, early-onset AD (EOAD) and very early-onset AD (VEOAD). The last two subtypes are associated with familial AD mutations. Clinical information is available concerning age and gender, age of onset and duration of disease, course and symptoms of the disease, medication and ApoE genotype. In the EOAD and VEOAD cases genetic testing was performed for APP, PSEN 1, PSEN2 and tau. For most cases, biomarkers in CSF and neuroimaging results are available. The isolated amyloid will serve as seeds to nucleate fibril growth for in vitro studies. The structure and polymorphism of the fibrils will be assessed by complementary structural approaches including solid-state NMR spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, and atomic force microscopy. Using the amyloid isolated from the five different subtypes of AD, we will assess the biofunctional consequences of the different strains using three approaches. First, we will assess the differences in the inflammatory response and cell toxicity due to different fibril forms using microglial cell cultures. Second, we will determine the influence of amyloid strains on promoting neuroinflammation. Third, we will determine the influence of amyloid strains on assembly and propagation in rat brain. The overall objective is to correlate pathologies (biofunctional consequences) of different amyloid subtypes between cell culture, rat brain and human brain, and to relate these pathologies with specific structural characteristics of the Aβ fibrils that are associated with the isolated amyloid from each subtype.

阿尔茨海默病与衰老过程中大脑中淀粉样蛋白的沉积有关。自从 淀粉样蛋白形成与 AD 之间的相关性最初被提出，现已认识到存在 该疾病可以采取多种亚型和形式。例如，β-淀粉样蛋白 (Aβ) 的积累 大脑实质中的缺陷是阿尔茨海默病 (AD) 的标志。尽管如此，还是有一个可怜的 了解淀粉样蛋白形成的原因，目前尚不清楚是否存在独特的结构基序 促进导致痴呆的独特病理后果。本提案的重点是填补这一空白 知识中的严重空白。因此，该提案的总体假设是 Aβ 肽形成 具有不同亚型的淀粉样蛋白具有决定其位置和病理学的独特结构。致地址 这个假设我们提出了两个具体目标。 首先，我们计划从迟发性 AD 和早期 AD 死后脑组织的不同亚型中分离淀粉样蛋白。 家族性AD发病。我们计划比较五种不同的淀粉样斑块亚型：典型 AD、非典型 AD、棉花 羊毛、早发性 AD (EOAD) 和极早发性 AD (VEOAD)。最后两个子类型与 家族性 AD 突变。可获得有关年龄和性别、发病年龄和持续时间的临床信息 疾病、病程和症状、药物治疗和 ApoE 基因型。在 EOAD 和 VEOAD 中 对 APP、PSEN 1、PSEN2 和 tau 病例进行基因检测。在大多数情况下，脑脊液中的生物标志物 和神经影像学结果可用。分离的淀粉样蛋白将作为种子使原纤维生长成核 体外研究。原纤维的结构和多态性将通过互补结构来评估 方法包括固态核磁共振光谱、傅里叶变换红外光谱、透射 电子显微镜、原子力显微镜。 使用从 AD 的五种不同亚型中分离出的淀粉样蛋白，我们将评估其生物功能 使用三种方法分析不同菌株的后果。首先，我们将评估差异 使用小胶质细胞培养物研究由于不同原纤维形式引起的炎症反应和细胞毒性。第二，我们 将确定淀粉样蛋白菌株对促进神经炎症的影响。第三，我们将确定 淀粉样蛋白菌株对大鼠脑中组装和传播的影响。 总体目标是关联不同淀粉样蛋白亚型的病理学（生物功能后果） 细胞培养物、大鼠大脑和人脑之间的关系，并将这些病理与特定的结构联系起来 与每个亚型中分离的淀粉样蛋白相关的 Aβ 原纤维的特征。