STRUCTURAL BIOLOGY OF ALPHA-SYNUCLEIN IN LEWY BODY DEMENTIA

路易体痴呆中α-突触核蛋白的结构生物学

基本信息

批准号：
10473717
负责人：
PAUL T KOTZBAUER
金额：
$ 80.57万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-30 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10473717
关键词：
Affinity Age of Onset Amino Acids Amyloid beta-Protein Antibodies Autopsy Binding Binding Sites Biological Assay Bradykinesia Brain Brain region Clinical Competitive Binding Cryo-electron tomography Cryoelectron Microscopy Data Dementia Dementia with Lewy Bodies Deposition Development Disease Disease Progression Electron Microscopy Elements Genes Genetic Polymorphism Goals Growth Human Imaging ligands Impairment In Vitro Individual Joints Kinetics Lewy Bodies Lewy Body Dementia Lewy neurites Ligand Binding Ligands Modeling Molecular Conformation Monoclonal Antibodies Motor Mutagenesis Mutation Neocortex Neurons Parkinson Disease Parkinson&apos s Dementia Pathogenesis Pathologic Polymorph Positron-Emission Tomography Posture Proteins Recombinants Reflex action Resolution Role Sampling Seeds Sodium Chloride Specificity Structure Subgroup Testing Therapeutic Time Tissue Sample Tissues Translations Tremor Variant Vertebral column abeta accumulation alpha synuclein base beta pleated sheet biomarker development brain tissue clinical diagnosis conformer illness length imaging agent improved monomer motor symptom new growth solid state nuclear magnetic resonance structural biology targeted biomarker targeted imaging targeted treatment tool

项目摘要

Parkinson’s disease (PD) is defined pathologically by the accumulation of alpha-synuclein (Asyn) fibrils in neuronal cytoplasmic and neuritic inclusions known as Lewy bodies and Lewy neurites. The role of Asyn in the pathogenesis of PD is supported by the identification of dominant mutations in the gene encoding Asyn (SNCA) in rare familial versions of PD. Dementia occurs frequently in PD. It sometimes begins at approximately the same time as motor symptoms (often referred to as dementia with Lewy bodies or DLB), or up to 20 years after motor symptoms begin (PD with dementia or PDD). The term Lewy body dementia (LBD) encompasses this spectrum of clinical presentations and is associated with widespread deposition of Asyn fibrils throughout the brain, particularly neocortex. Multiple therapeutic approaches targeting Asyn accumulation are being pursued. A further priority is to develop a PET imaging agent to quantify the deposition of Asyn in living individuals, as a biomarker for target engagement and disease progression. Understanding Asyn fibril structure in LBD can guide the development of Asyn-targeted therapies and imaging agents. In this project, we will use cryo-electron microscopy (cryo-EM) to determine atomic resolution structures of Asyn fibrils in LBD, in conjunction with solid-state NMR (SSNMR) for refinement of structures. We will analyze and compare structures of Asyn fibrils isolated from multiple subgroups of LBD autopsy cases defined by early versus late onset of dementia, as well as the presence or absence of co-occurring amyloid β accumulation. We will also utilize cryo-electron tomography, SSNMR spectral analysis and new monoclonal antibodies to extend the analysis of Asyn fibril structure to additional autopsy cases. To promote the translation of these structural studies we will utilize cryo-EM to determine binding sites of leading candidates for PET imaging ligand and use fibril growth assays to identify specific amino acid residues in the Asyn protein that are important for fibril growth and stability.

帕金森氏病（PD）在病理上是通过神经元细胞质和神经性夹杂物中的α-核蛋白（ASYN）原纤维的积累来定义的。 ASYN在PD发病机理中的作用得到了鉴定在PD的稀有家族版本中编码ASYN（SNCA）的基因中的显性突变。痴呆症经常出现在PD中。有时它与运动症状（通常称为Lewy尸体或DLB的痴呆症）或运动症状开始后长达20年（伴有痴呆或PDD的PD）大约开始。 Lewy身体痴呆（LBD）一词涵盖了这一临床表现的范围，并与整个大脑中的Asyn原纤维的宽度沉积有关，尤其是新皮层。正在追求针对ASYN积累的多种治疗方法。另一个优先级是开发宠物成像剂来量化ASYN在活人中的沉积，作为目标参与和疾病进展的生物标志物。了解LBD中的Asyn Fibril结构可以指导以ASYN为目标的疗法和成像剂的发展。在该项目中，我们将使用低温电子显微镜（Cryo-EM）确定LBD中Asyn原纤维的原子分辨率结构，并与固态NMR（SSNMR）结合结合结构。我们将分析和比较从痴呆症早期和晚期发作定义的LBD尸检病例的多个亚组中分离出的ASYN原纤维的结构，以及存在或不存在淀粉样βββ的积累。我们还将利用冷冻电子断层扫描，SSNMR光谱分析和新的单克隆抗体将ASYN原纤维结构的分析扩展到其他尸检病例。为了促进这些结构研究的翻译，我们将利用冷冻EM来确定宠物成像配体的主要候选物的结合位点，并使用原纤维生长测定法确定在Asyn蛋白中保留的特定氨基酸对原纤维生长和稳定性很重要的氨基酸。