Heterotypic amyloid interactions as modulators of selective cellular vulnerability

异型淀粉样蛋白相互作用作为选择性细胞脆弱性的调节剂

• 批准号: 10524877
• 负责人: Frederic Rousseau
• 金额: $ 52.37万
• 依托单位: FLANDERS INTERUNIV INST BIOTECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10524877
• 关键词: Affect; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid; Amyloid beta-Protein; Animals; Binding; Biological; Biological Assay; Biophysics; Biotechnology; Brain; Brain region; Cell Culture System; Cell Line; Cell model; Cells; Complex; Data Set; Deposition; Disease; Event; Fluorescence Resonance Energy Transfer; In Situ; In Vitro; Institutes; Laboratories; Lead; Length; Mediating; Modeling; Molecular; Morphology; Neurons; Pathologic; Pathology; Play; Proteins; Proteome; Proteomics; Recombinant adeno-associated virus (rAAV); Reporter; Research; Research Project Grants; Role; Running; Sequence Homology; Set protein; Site; Software Tools; Specificity; Statistical Data Interpretation; Structural Models; Structure; Therapeutic; Therapeutic Effect; Viral; abeta accumulation; abeta deposition; amyloid formation; amyloid pathology; base; biophysical techniques; brain cell; computational pipelines; extracellular; in silico; in vivo; mouse model; overexpression; proteostasis; tau Proteins; tau aggregation; vector

ABSTRACT Amyloid aggregates are the defining pathological hallmark of Alzheimer’s Disease (AD), yet the role they play and the therapeutic effect in targeting these aggregates remains controversial. Little is known about the impact of the proteome context in which these proteins reside, or what nucleates their aggregation in specific sites in the brain. Studying the composition of amyloid deposits using proteomic approaches has demonstrated the co- deposition of many other proteins, however currently there is no straightforward chain-of-events that explains plaque composition. The predicament in which the field currently subsists critically highlights the lack of suitable structural-mechanistic models to understand both the causes and consequences of amyloid aggregation in terms of direct molecular interactions, as well as which specific cellular factors determine pathognomonic disease initiation. In this project, the Switch Laboratory in VIB Flanders Institute for Biotechnology will approach the selective amyloid aggregation of beta amyloid (Aβ) and tau in AD mechanistically and will do a systematic search for potential interacting partners based on the sequence- and structure-specificity of aggregation. This systematic and proteome-wide screen is based on the assumption that amyloid aggregation is initiated by the specific interaction of aggregation-prone regions (APRs) within Aβ and tau with aggregation-prone sequence segments in other proteins within the background proteome. They have developed a unique computational pipeline to model heterotypic interactions with sufficient predictive power to identify amyloid modifiers in cells. The project will investigate the in vivo impact of heterotypic amyloid interactions in mouse models and for the hits, will analyze in molecular detail how the aggregation of Aβ and tau is modified by the interactions. Aim 1 will run an in-silico screen with special emphasis on known factors related to selective vulnerably. The computational screen will use all-atom modelling of sequence segments with sequence homology to the APRs of Aβ and tau to identify other brain-expressed proteins that may modify the aggregation of Aβ or tau. Aim 2 will screen full-length proteins in cellular models to identify candidates that can modify amyloid aggregation of Aβ and tau in a complex biological context. Aim 3 will identify modifiers that have an effect in vivo by expressing the most potent modifier proteins in mouse models, to study the impact on aggregation onset and extent of amyloid pathology of Aβ and tau. For the most promising modifiers identified in Aims 2 and 3, Aim 4 will unravel the molecular mechanism of selected heterotypic amyloid interactions use state-of-the-art biophysical methods to elucidate exactly how these interactions change the amyloid formation of Aβ and tau.

抽象的 淀粉样蛋白聚集体是阿尔茨海默氏病（AD）的定义病理标志，但它们扮演的角色 靶向这些骨料的治疗作用仍然存在争议。对影响知之甚少 这些蛋白质所在的蛋白质含量，或在特定位点的核核中 大脑。使用蛋白质组学方法研究淀粉样蛋白沉积物的组成表明 许多其他蛋白质的沉积，但是目前没有直接的事件可以解释 斑块组成。该领域当前订阅者的预测批评不足 结构机械模型以理解淀粉样蛋白聚集的原因和后果 直接分子相互作用以及哪些特定细胞因子决定了病原体疾病 引发。 在这个项目中，Vib Flanders生物技术研究所的开关实验室将接近选择性 β-淀粉样β淀粉样蛋白（Aβ）和tau的淀粉样蛋白聚集在机械上，将进行系统的搜索 潜在的相互作用伙伴基于聚合的序列和结构特异性。这个系统 蛋白质组范围的屏幕基于以下假设：淀粉样蛋白聚集是由特定的 Aβ和TAU内聚集区域（APR）与易于聚集的序列段的相互作用 在背景蛋白质组中的其他蛋白质中。 他们开发了独特的计算管道，以建模具有足够预测性的异型相互作用 鉴定细胞中淀粉样蛋白修饰剂的功率。该项目将研究异型淀粉样蛋白的体内影响 小鼠模型中的相互作用和点击中，将分子细节分析Aβ和TAU的聚集如何 通过相互作用修改。 AIM 1将运行一个内部屏幕，特别强调与选择性相关的已知因素。这 计算屏幕将使用具有序列同源性的序列段的全原子建模 Aβ和TAU的鉴定，可以鉴定其他脑表达的蛋白质，这些蛋白可能会改变Aβ或TAU的聚集。 AIM 2意志 筛选细胞模型中的全长蛋白质，以鉴定可以改变Aβ淀粉样蛋白聚集的候选者 和tau在复杂的生物学环境中。 AIM 3将通过表达在体内产生影响的修饰符 小鼠模型中的大多数锅修饰蛋白，以研究对淀粉样蛋白聚集的影响和程度的影响 Aβ和TAU的病理。对于目标2和3中确定的最有前途的修饰符，AIM 4将揭示 选定的异型淀粉样相互作用的分子机制使用最先进的生物物理方法 阐明这些相互作用如何改变Aβ和TAU的淀粉样蛋白形成。