Structures and Propagation of Pathologically Relevant Amyloids in Alzheimer's

阿尔茨海默病中病理相关淀粉样蛋白的结构和传播

基本信息

批准号：
9927626
负责人：
YOSHITAKA ISHII
金额：
$ 28.65万
依托单位：
TOKYO INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-01 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9927626
关键词：
Address Affect Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease diagnosis Amino Acid Sequence Amyloid Amyloid Fibrils Amyloid beta-42 Amyloid beta-Protein Attention Biochemical Brain Caliber Cell Death Cessation of life Chemicals Communities Diffuse Disease Progression Fingerprint Fluorescence Freezing Homo Human Immune In Vitro Infection Kinetics Link Methods Minor Modeling Molecular Structure Nerve Degeneration Neuronal Dysfunction Neurons Neurotoxins Pathogenicity Pathologic Patients Play Preparation Presenile Alzheimer Dementia Prions Property Publishing Role Sampling Senile Plaques Severities Site Spectrum Analysis Structure Time Toxic effect Transmission Electron Microscopy Variant X-Ray Crystallography abeta oligomer amyloid structure beta pleated sheet biophysical techniques experimental study in vitro Model insight mimetics mutant novel strategies prion-like public health relevance reconstitution relating to nervous system self assembly simulation solid state nuclear magnetic resonance tool

项目摘要

DESCRIPTION (provided by applicant): Amyloid fibrils of 42- and 40-residues Alzheimer's β-amyloid (Aβ) peptides represent two primary components of senile plaques, which are a hallmark of Alzheimer's disease (AD). As amyloid fibrils formed via self-assembly of Aβ show notable toxicity, neural cell deaths in AD have long been associated with Aβ amyloid fibrils. More recently, diffusible subfibrillar aggregates of Aβ were identified as a new suspect of AD since many of these diffusible Aβ aggregates show much higher toxicity than Aβ fibrils. Molecular structures of amyloid fibrils and diffusible aggregates of Aβ have attracted great attention. However, applications of traditional methods such as X-ray crystallography or solution NMR have been limited to non-crystalline amyloid aggregates. In particular, for more pathologically relevant 42-residue Aβ (Aβ42), only a few atomic-level structural details were known because of notoriously difficult sample preparation due to its high propensity to aggregate. Similarly, for Aβ mutants associated with early onset of AD, very little is known about structures of most pathogenic mutants such as E22G Aβ because they aggregate readily into heterogeneous aggregates, which are not suited for a structural analysis. Through this study, we will reveal atomic details of amyloid fibrils and spherical aggregates in order to highlight structural relationships and interactions between different amyloid aggregate species, in particular for Aβ42, which is considered to be more pathogenic than more abundant 40-residue Aβ40. As a vital tool for site- specific structural analysis for amyloid aggregates, we will use solid-state NMR (SSNMR), in a combination with transmission electron microscopy (TEM), MD simulations, and other biochemical/biophysical methods. Our studies entail the following three Specific Aims. (1) In Aim 1, we will establish preparation of homogenous amyloid fibrils optimized for structural analyses for Aβ42 and a pathogenic mutant E22G Aβ40. For the Aβ42 fibrils, we will elucidate atomic-level structural details by advanced SSNMR spectroscopy. The interactions of different Aβ species in AD will be modeled by cross-seeding experiments, which allow us to examine structural compatibility of two different Aβ species in fibrils. The aim will define structures and structural variations of Aβ42 fibrils while offering insight into how the structures can impact AD progression. (2) In Aim 2, we will examine atomic details of two distinctive diffusible Aβ42 aggregates. First, we will establish a synthetic reconstitute of highl toxic subfibrillar aggregate called amylospheroid (ASPD), which is found in AD brains. As the level of ASPD is correlated with the severity of AD, this study will reveal the first atomic detail a pathologically relevant amyloid oligomer in AD. We will also examine a similar spherical assembly called SPA, which has a larger size (20 nm diameter) and modest toxicity. (3) In Aim 3, we will profile structures and toxicity of brain-derived Aβ42 fibrils, which are replicated fro Aβ42 fibrils in AD brain. Through a comparison of ~30 brain samples, the aim will allow us to compare structures and toxicity of brain-derived amyloid fibrils in AD.

描述（由适用提供）：42个和40个沉积物阿尔茨海默氏症的β-淀粉样蛋白（Aβ）肽的淀粉样蛋白原纤维代表了老年斑块的两个主要成分，这是阿尔茨海默氏病（AD）的标志。由于通过Aβ自组装形成的淀粉样蛋白原纤维表现出显着的毒性，因此AD中的神经细胞死亡长期与Aβ淀粉样蛋白原纤维有关。最近，Aβ的扩散亚纤维聚集体被确定为AD的新嫌疑犯，因为这些扩散的Aβ聚集体中的许多毒性表现出比Aβ原纤维高得多的毒性。淀粉样蛋白原纤维和Aβ的扩散骨料的分子结构引起了极大的关注。但是，X射线晶体学或溶液NMR等传统方法的应用仅限于非晶淀粉样蛋白聚集体。特别是，对于更具病理相关的42个残留Aβ（Aβ42），由于众所周知的样品制备了众所周知的较高的样品准备，因此仅知道少数原子级的结构细节。同样，对于与AD早期发作相关的Aβ突变体，关于大多数致病突变体（例如E22GAβ）的结构很容易聚集成异质骨料，这些聚集体不适合结构分析。通过这项研究，我们将揭示淀粉样蛋白原纤维和球形骨料的原子细节，以突出不同淀粉样蛋白聚集体之间的结构关系和相互作用，特别是对于Aβ42，这被认为是比40多种残留的Aβ40更具致病性的。作为淀粉样蛋白聚集体的位点特异性结构分析的重要工具，我们将与透射电子显微镜（TEM），MD模拟和其他生化/生物物理学方法结合使用固态NMR（SSNMR）。我们的研究需要以下三个具体目标。（1）在AIM 1中，我们将建立针对Aβ42和致病性突变体E22GAβ40的结构分析优化的同质淀粉样蛋白原纤维的制备。对于Aβ42纤维，我们将通过晚期SSNMR光谱阐明原子级的结构细节。 AD中不同Aβ物种的相互作用将通过交叉种子实验来建模，这使我们能够检查原纤维中两个不同Aβ物种的结构兼容性。该目标将定义Aβ42原纤维的结构和结构变化，同时洞悉结构如何影响AD的进展。（2）在AIM 2中，我们将检查两个独特的扩散Aβ42聚集体的原子细节。首先，我们将建立一个称为Amylospheroid（ASPD）的高毒性亚纤维骨料的合成重构，该聚集体在AD大脑中发现。由于ASPD的水平与AD的严重程度相关，因此该研究将揭示AD中第一个原子细节与病理相关的淀粉样蛋白低聚物。我们还将检查一个称为水疗中心的类似球形组件，该组件的尺寸较大（20 nm）和适度的毒性。（3）在AIM 3中，我们将介绍大脑衍生的Aβ42原纤维的结构和毒性，这些原纤维在AD脑中被复制到Aβ42纤维中。通过比较约30个大脑样本，其目的将使我们能够比较AD中脑源性淀粉样蛋白纤维的结构和毒性。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Controlled functionalization of graphene oxide with sodium azide.

DOI：
10.1039/c3nr04332k
发表时间：
2013-12-21
期刊：
Nanoscale
影响因子：
6.7
作者：
Eigler S;Hu Y;Ishii Y;Hirsch A
通讯作者：
Hirsch A

Aβ(1-42) fibril structure illuminates self-recognition and replication of amyloid in Alzheimer's disease.

DOI：
10.1038/nsmb.2991
发表时间：
2015-06
期刊：
NATURE STRUCTURAL & MOLECULAR BIOLOGY
影响因子：
16.8
作者：
Xiao, Yiling;Ma, Buyong;McElheny, Dan;Parthasarathy, Sudhakar;Long, Fei;Hoshi, Minako;Nussinov, Ruth;Ishii, Yoshitaka
通讯作者：
Ishii, Yoshitaka

Solid-State NMR Studies of Amyloid Materials: A Protocol to Define an Atomic Model of Aβ(1-42) in Amyloid Fibrils.

淀粉样蛋白材料的固态核磁共振研究：定义淀粉样原纤维中 Aβ(1-42) 原子模型的协议。