Structure and conformational diversity of amyloid oligomers

淀粉样蛋白寡聚物的结构和构象多样性

• 批准号: 7897965
• 负责人: Charles G. Glabe
• 金额: $ 27.92万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7897965
• 关键词: Address; Affinity; Alzheimer' s Disease; Amino Acid Sequence; Amyloid; Amyloid Fibrils; Amyloid Proteins; Antibodies; Antibody Specificity; Asses; Binding; Brain; Brain Diseases; Cells; Classification; Cognitive; Conflict (Psychology); Data; Deposition; Development; Disease; Epitopes; Face; Generic Drugs; Goals; Heterogeneity; Higher Order Chromatin Structure; Human; Huntington Disease; Immune Sera; Impaired cognition; In Vitro; Laboratories; Lead; Ligands; Molecular Conformation; Monitor; Monoclonal Antibodies; Mutation; Neurodegenerative Disorders; Neurons; Parkinson Disease; Pathogenesis; Pathology; Pathway interactions; Peptides; Pharmaceutical Preparations; Play; Preparation; Presenile Alzheimer Dementia; Principal Investigator; Prion Diseases; Protein Precursors; Proteins; Publishing; Rattus; Reagent; Reporting; Research Project Grants; Role; Running; Sampling; Seeds; Side; Specificity; Structure; Testing; Toxic effect; Transgenic Mice; Variant; Vertebral column; aggregation pathway; amyloid structure; base; cellular targeting; conformer; insight; monomer; prevent; programs; public health relevance; small molecule; synthetic peptide; therapeutic development; therapeutic target; three dimensional structure; Address; Affinity; Alzheimer' s Disease; Amino Acid Sequence; Amyloid; Amyloid Fibrils; Amyloid Proteins; Antibodies; Antibody Specificity; Asses; Binding; Brain; Brain Diseases; Cells; Classification; Cognitive; Conflict (Psychology); Data; Deposition; Development; Disease; Epitopes; Face; Generic Drugs; Goals; Heterogeneity; Higher Order Chromatin Structure; Human; Huntington Disease; Immune Sera; Impaired cognition; In Vitro; Laboratories; Lead; Ligands; Molecular Conformation; Monitor; Monoclonal Antibodies; Mutation; Neurodegenerative Disorders; Neurons; Parkinson Disease; Pathogenesis; Pathology; Pathway interactions; Peptides; Pharmaceutical Preparations; Play; Preparation; Presenile Alzheimer Dementia; Principal Investigator; Prion Diseases; Protein Precursors; Proteins; Publishing; Rattus; Reagent; Reporting; Research Project Grants; Role; Running; Sampling; Seeds; Side; Specificity; Structure; Testing; Toxic effect; Transgenic Mice; Variant; Vertebral column; aggregation pathway; amyloid structure; base; cellular targeting; conformer; insight; monomer; prevent; programs; public health relevance; small molecule; synthetic peptide; therapeutic development; therapeutic target; three dimensional structure

DESCRIPTION (provided by applicant): The goal of this research project is to explore the structural heterogeneity of amyloid aggregates and the relationships of this conformational variation to the toxicity or pathogenic activities of amyloid oligomers. Several distinct types of amyloid deposits accumulate in disease brain and current evidence suggests that soluble, oligomeric forms of Ass may play primary role in pathogenesis. Recent results indicate that conformation-dependent monoclonal antibodies can distinguish between different types soluble Ass oligomers. These antibodies also distinguish other conformations of Ass, including monomers, fibrils and natively-folded APP. We have recently prepared two additional conformation-dependent antisera that recognize generic epitopes associated with amyloid fibrils and pore-like annular protofibrils that are formed from many different types of amyloids (see Preliminary Data, below). These antibodies are complementary to the anti-oligomer antibody, A11 and recognize epitopes that are specific to fibrils and annular protofibril aggregates. We hypothesize that these conformationally distinct assembly states of Ass are differentially associated with AD pathogenesis. We anticipate that these results will help clarify some apparent inconsistencies and conflicting data, such as the observations that the total Ass amyloid deposited correlates poorly with disease and some people have large amounts of amyloid and are cognitive normal, while other brain samples that have little observable amyloid deposits are associated with cognitive dysfunction. We hypothesize that the distinct types of soluble oligomeric or annular protofibril forms of Ass have distinct toxicities. Conformation-dependent antibodies hold the potential of identifying and distinguishing specific assembly states because they only recognize a specific misfolded state and do not react with the natively folded precursor protein. The specific aims of this project will address the following questions: What is the conformational diversity of amyloid aggregates? What is the structural basis of conformation dependent antibody specificity and amyloid oligomer structural? What are the relationships between different conformational states of amyloids? What is the pathological significance of the different amyloid conformational states? The answers to these questions should provide insight into the range of amyloid oligomer conformational diversity and monoclonal antibody reagents that distinguish different conformations of oligomers. This may provide a more rational structural basis for the classification of oligomers and provide insight into the variability in oligomer preparations reported by different laboratories. Determining the 3 dimensional structure of the monoclonal Fabs bound to amyloid oligomers may provide unprecedented insight into the structure of amyloid oligomers and the mechanism of specific antibody recognition that may be useful for development of immunological therapeutics that target oligomer formation or prevent their interaction with cellular targets. The identification of small molecules that specifically inhibit the formation of different types of oligomers should help to clarify whether the oligomers are intermediates in the formation of higher order structures, like fibrils or whether they represent stable alternative end products. This aim may also provide small molecule lead compounds that specifically inhibit amyloid oligomer formation for therapeutic development. The characterization of which types of amyloid oligomers are more closely related to pathogenesis in human and transgenic mouse brain may help to identify targets for therapeutic development. PHS 398/2590 (Rev. 04/06) Page Continuation Format Page PUBLIC HEALTH RELEVANCE: Amyloid aggregates are believed to play a central role in the development of neurodegenerative diseases, like Alzheimer, Parkinson, Huntington and prion diseases. The goal of this proposal is to determine how many different types of amyloid aggregates exist and to determine the precise 3 dimensional structure of the amyloid aggregates. The proposal will also examine which of these aggregates is more toxic to neurons and most closely related to disease. An additional benefit is that this proposal may produce monoclonal antibodies and drugs that specifically target these amyloid aggregates for therapeutic development.

描述（由申请人提供）：该研究项目的目的是探索淀粉样蛋白聚集体的结构异质性以及这种构型变化与淀粉样蛋白寡聚物的毒性或致病活性的关系。几种不同类型的淀粉样蛋白沉积物积累在疾病大脑中，目前的证据表明，可溶的，寡聚形式的ASS可能在发病机理中起主要作用。最近的结果表明，依赖构象的单克隆抗体可以区分不同类型的可溶性屁股低聚物。这些抗体还区分了屁股的其他构象，包括单体，原纤维和折叠的应用。我们最近准备了两个额外的构型依赖性抗血清，它们识别与淀粉样蛋白纤维和孔状的环形原子纤维相关的通用表位，它们是由许多不同类型的淀粉样蛋白形成的（请参阅下面的初步数据）。这些抗体与抗橄榄抗体A11互补，并识别特有的纤维和环形原纤维聚集体的表位。我们假设这些构型不同的ASS组装状态与AD发病机理有差异化。我们预计这些结果将有助于阐明一些明显的不一致和数据相互矛盾的数据，例如观察到，总淀粉样蛋白沉积的总淀粉样蛋白与疾病的相关性很差，有些人具有很大的淀粉样蛋白，并且具有很少的淀粉样蛋白沉积物的其他脑样本，而淀粉样蛋白的沉积物几乎没有与认知能力相关。我们假设可溶性的寡聚或环形原纤维形式的不同类型的ASS具有不同的毒性。构型依赖性抗体具有识别和区分特定组装状态的潜力，因为它们仅识别特定的错误折叠状态，并且不会与本质上折叠的前体蛋白反应。该项目的具体目的将解决以下问题：淀粉样蛋白聚集体的构象多样性是什么？构型依赖性抗体特异性和淀粉样蛋白低聚物结构的结构基础是什么？淀粉样蛋白不同构象状态之间有什么关系？不同淀粉样蛋白构象状态的病理意义是什么？这些问题的答案应提供有关淀粉样蛋白低聚物构象多样性和单克隆抗体试剂的范围，从而区分了低聚物的不同构象。这可以为低聚物的分类提供更合理的结构基础，并洞悉不同实验室报告的低聚物制剂的变异性。确定与淀粉样蛋白低聚物结合的单克隆晶圆剂的3维结构可能会提供对淀粉样蛋白低聚物结构的前所未有的见解，以及特定抗体识别的机制，这些机制可能有用，可用于开发具有靶向寡聚物或防止其与细胞靶标相互作用的免疫学治疗剂的发展。特异性抑制不同类型的低聚物形成的小分子的鉴定应有助于阐明寡聚物在高阶结构形成中是否是中间体，例如原纤维，还是代表稳定的替代最终产品。该目标还可以提供小分子铅化合物，这些化合物专门抑制淀粉样蛋白寡聚物形成以进行治疗性发育。哪种类型的淀粉样蛋白低聚物的表征与人和转基因小鼠脑的发病机理更紧密相关，可能有助于识别治疗性发育的靶标。 PHS 398/2590（修订版04/06）页面延续格式页面 公共卫生相关性：据信淀粉样蛋白骨料在神经退行性疾病的发展中起着核心作用，例如阿尔茨海默氏症，帕金森氏症，亨廷顿和普里奥疾病。该建议的目的是确定存在多少种不同类型的淀粉样蛋白聚集体，并确定淀粉样蛋白聚集体的精确3维结构。该提案还将检查哪些聚集体对神经元更具毒性，并且与疾病最密切相关。另一个好处是，该建议可能会产生单克隆抗体和药物，这些抗体和药物专门针对这些淀粉样蛋白骨料用于治疗性发育。