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

• 批准号: 10729722
• 负责人: PAUL T KOTZBAUER
• 金额: $ 423.15万
• 依托单位: WASHINGTON UNIVERSITY
• 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 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10729722
• 关键词: Address; Advanced Development; Affect; Affinity; Amino Acids; Amygdaloid structure; Amyloid beta-Protein; Animal Model; Autopsy; Binding; Binding Sites; Biological Markers; Biological Models; Bradykinesia; Brain; Brain region; Cell Culture Techniques; Cells; Clinical; Complement; Corpus striatum structure; Cryo-electron tomography; Cryoelectron Microscopy; Cytoplasm; Dementia; Dementia with Lewy Bodies; Deposition; Development; Disease; Disease Progression; Elements; Future; Genes; Goals; Growth; Imaging ligands; Impairment; In Situ; In Vitro; Individual; Injections; Isotope Labeling; Kinetics; Label; Lead; Lewy Bodies; Lewy Body Dementia; Lewy neurites; Ligand Binding; Ligands; Methods; Modeling; Motor; Mus; Neocortex; Neurons; Parkinson Disease; Parkinson' s Dementia; Pathogenesis; Pathologic; Polymorph; Positron-Emission Tomography; Posture; Proteins; Recombinants; Reflex action; Regulation; Reporting; Resolution; Role; Solvents; Structural Models; Structure; Subgroup; Therapeutic; Time; Tissue Sample; Tissues; Translations; Tremor; Variant; abeta accumulation; alpha synuclein; biomarker development; brain tissue; clinical diagnosis; clinical phenotype; conformer; design; disease mechanisms study; disease phenotype; dominant genetic mutation; electron tomography; imaging agent; improved; innovation; insight; milligram; monomer; motor symptom; mouse model; nano; nanomolar; neocortical; particle; solid state nuclear magnetic resonance; structural biology; synuclein; targeted biomarker; targeted imaging; targeted treatment; therapeutic development

Abstract Parkinson 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 Positron Emission Tomography (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 a combination of solid-state NMR (SSNMR) and cryo- electron microscopy (cryo-EM) to determine atomic resolution structures of Asyn fibrils in LBD. We developed multiple complimentary approaches to isolate fibrils from tissue and grow them in the presence of labeled monomeric Asyn protein for SSNMR and cryo-EM studies, enabling more comprehensive analysis of structure. 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, and determine whether structural variations relate to disease phenotype. To promote the translation of these structural studies we will utilize SSNMR and cryo-EM to determine the structural features of binding sites for leading PET imaging ligand candidates, which can guide further optimization of PET ligands. Finally, we will utilize Asyn fibrils derived from LBD tissue to seed accumulation of Asyn fibrils in cell culture and mouse models. We will utilize SSNMR, cryo-EM and cryo-electron tomography to analyze Asyn fibril structure in these model systems, which will guide future studies of disease mechanisms, PET ligand development and therapeutic development.

抽象的 帕金森病 (PD) 的病理学定义为 α-突触核蛋白 (Asyn) 原纤维的积累 神经元细胞质和神经炎包涵体称为路易体和路易神经突。 Asyn 在其中的作用 编码 Asyn 的基因中显性突变的鉴定支持了 PD 的发病机制 (SNCA) 罕见的家族性帕金森病。痴呆症常发生在帕金森病中。有时开始于 与运动症状（通常称为路易体痴呆或 DLB）大约同时发生，或 运动症状开始后最多 20 年（PD 伴痴呆或 PDD）。路易体痴呆 (LBD) 一词 涵盖了这一系列的临床表现，并与 Asyn 的广泛沉积有关 纤维遍布整个大脑，特别是新皮质。针对 Asyn 的多种治疗方法 正在追求积累。进一步优先考虑的是开发正电子发射断层扫描 (PET) 成像剂来量化活体中 Asyn 的沉积，作为目标参与的生物标志物 疾病进展。了解 LBD 中的 Asyn 原纤维结构可以指导 Asyn 靶向的开发 疗法和显像剂。在这个项目中，我们将结合使用固态核磁共振 (SSNMR) 和冷冻核磁共振技术。 电子显微镜 (cryo-EM) 确定 LBD 中 Asyn 原纤维的原子分辨率结构。我们开发了 多种互补方法从组织中分离原纤维并在标记的存在下生长它们 用于 SSNMR 和冷冻电镜研究的单体 Asyn 蛋白，可实现更全面的结构分析。 我们将分析和比较从 LBD 尸检病例的多个亚组中分离出的 Asyn 原纤维的结构 定义为痴呆的早发与晚发，以及是否存在同时存在的β淀粉样蛋白 积累，并确定结构变异是否与疾病表型相关。为促进 这些结构研究的转化，我们将利用 SSNMR 和冷冻电镜来确定 主要 PET 成像配体候选物的结合位点，可以指导 PET 配体的进一步优化。 最后，我们将利用来自 LBD 组织的 Asyn 原纤维在细胞培养物中进行 Asyn 原纤维的种子积累 和鼠标模型。我们将利用 SSNMR、冷冻电镜和冷冻电子断层扫描来分析 Asyn 原纤维 这些模型系统中的结构，这将指导疾病机制、PET配体的未来研究 的发展和治疗的发展。