Production and Distribution of well-characterized polymorphic variants of alpha-synuclein aggregates

α-突触核蛋白聚集体的充分表征的多态性变体的生产和分布

• 批准号: 10706583
• 负责人: CLAUDIO SOTO
• 金额: $ 75.95万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 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/10706583
• 关键词: Adopted; Affect; Alzheimer' s Disease; Alzheimer' s disease patient; Alzheimer' s disease related dementia; Amyloid beta-Protein; Animal Model; Animals; Appearance; Biochemical; Biological; Biological Assay; Biophysics; Brain; Cell model; Cells; Characteristics; Communities; Cryoelectron Microscopy; Dementia; Dementia with Lewy Bodies; Deposition; Disease; Documentation; Ensure; Exhibits; Exposure to; Freezing; Goals; Guidelines; Human; In Vitro; Investigation; Laboratories; Lewy Bodies; Lewy Body Dementia; Lewy neurites; Liquid substance; Methodology; Modeling; Molecular Conformation; Movement Disorders; Neurofibrillary Tangles; Neurons; Parkinson Disease; Parkinson' s Dementia; Pathologic; Pathology; Patients; Polymorph; Prevalence; Process; Production; Property; Proteins; Protocols documentation; Reporter; Reproducibility; Research; Research Personnel; Resolution; Resources; Role; Sampling; Senile Plaques; Structure; Technology; United States National Institutes of Health; Variant; alpha synuclein; amplification detection; biological systems; biophysical properties; clinical phenotype; in vivo; in vivo Model; misfolded protein; mouse model; neurodegenerative dementia; neurotoxic; protein aggregation; reagent standardization; response; synucleinopathy; tau Proteins; tau aggregation

ABSTRACT Alzheimer's disease (AD) is the most common form of dementia and may contribute to 60--70% of cases, followed in prevalence by Lewy body dementia (LBD), responsible for 17% of the cases. AD and related dementias are thought to be caused by the accumulation of misfolded protein aggregates in the brain. In AD, protein aggregates mostly appear in the form of amyloid plaques and neurofibrillary tangles composed of amyloid-beta (ABeta) and hyperphosporylated Tau, respectively. In LBD the main component of the aggregates is the alpha-synuclein (aSyn) protein. However, there is a substantial overlap of pathological abnormalities, leading to the relatively frequent appearance of mixed pathologies, characterized by the presence of multiple protein aggregates in the same brain. Indeed, up to 30-40% of AD patients display aSyn Lewy bodies in their brains. Although, much investigation has been devoted to understand the role of A~ and Tau aggregates in AD, little research has been done to evaluate the contribution of aSyn in AD and related dementias. Recently, we implemented and developed a highly sensitive and specific assay (termed aSyn-PMCA) to faithfully amplify aSyn seeds present in brain and biological fluids of patients affected by various synucleinopathies, including LBD. Using this technology we were able to amplify seeding competent aSyn aggregates from patients' samples and identify conformational strains associated to different diseases. The major goal of this project is to take advantage of the reliability of the aSyn-PMCA technology to massively amplify aSyn polymorphic variants in order to produce and distribute to the scientific community well-characterized aSyn aggregates amplified from patient's samples. Materials will be characterized for their size, biochemical, biophysical, structural and biological properties using various in vitro, cellular and animal models. Specific aims include: (1) Production of aSyn aggregates by aSyn-PMCA amplification from brain and CSF of patients affected by AD and LBD (as well as other synucleinopathies) and comprehensive analysis of the size, biochemical and biophysical properties of these aggregates; (2) Study the structural basis differentiating different synucleinopathies, in particular we propose to elucidate the structural fold at atomic resolution for aSyn aggregates implicated in different diseases; (3) Study the biological properties of aSyn aggregates derived from AD, LBD and other synucleinopathies using in vitro and in vivo models. (4) Produce and distribute to the scientific community well-characterized and biologically relevant aSyn aggregates. The resources generated in this project will be made fully available to researchers in order to standardize reagents and ensure the biological relevance of the findings.

抽象的 阿尔茨海默氏病 (AD) 是最常见的痴呆症，可能导致 60--70% 的病例，其次是路易体痴呆 (LBD)，占 17% 的病例。 AD 和相关痴呆症被认为是由大脑中错误折叠的蛋白质聚集体的积累引起的。在 AD 中，蛋白质聚集体主要以淀粉样斑块和神经原纤维缠结的形式出现，分别由淀粉样蛋白-β (ABeta) 和过度磷酸化的 Tau 组成。在 LBD 中，聚集体的主要成分是 α-突触核蛋白 (aSyn)。然而，病理异常存在大量重叠，导致混合病理相对频繁地出现，其特征是同一大脑中存在多种蛋白质聚集体。事实上，高达 30-40% 的 AD 患者大脑中存在 aSyn Lewy 小体。尽管已经进行了大量研究来了解 A~ 和 Tau 聚集体在 AD 中的作用，但很少有研究来评估 aSyn 在 AD 和相关痴呆中的作用。最近，我们实施并开发了一种高度敏感和特异性的测定（称为 aSyn-PMCA），以忠实地放大受各种突触核蛋白病（包括 LBD）影响的患者的大脑和生物体液中存在的 aSyn 种子。利用这项技术，我们能够从患者样本中扩增具有播种能力的 aSyn 聚集体，并鉴定与不同疾病相关的构象菌株。该项目的主要目标是利用 aSyn-PMCA 技术的可靠性来大规模扩增 aSyn 多态性变体，以便生产并向科学界分发从患者样本中扩增的特征良好的 aSyn 聚集体。将使用各种体外、细胞和动物模型来表征材料的尺寸、生化、生物物理、结构和生物学特性。具体目标包括： (1) 通过 aSyn-PMCA 扩增从 AD 和 LBD（以及其他突触核蛋白病）患者的大脑和 CSF 中产生 aSyn 聚集体，并对这些聚集体的大小、生化和生物物理特性进行综合分析； (2) 研究区分不同突触核蛋白病的结构基础，特别是我们建议以原子分辨率阐明不同疾病中涉及的 aSyn 聚集体的结构折叠； (3) 使用体外和体内模型研究源自 AD、LBD 和其他突触核蛋白病的 aSyn 聚集体的生物学特性。 (4) 生产并向科学界分发具有良好特征且具有生物学相关性的 aSyn 聚集体。该项目产生的资源将充分提供给研究人员，以标准化试剂并确保研究结果的生物学相关性。

项目成果

(De)stabilization of Alpha-Synuclein Fibrillary Aggregation by Charged and Uncharged Surfactants.

• DOI: 10.3390/ijms222212509
• 发表时间: 2021-11-19
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: Loureiro JA;Andrade S;Goderis L;Gomez-Gutierrez R;Soto C;Morales R;Pereira MC
• 通讯作者: Pereira MC