Differentiation and characterization of conformational alpha-synuclein strains associated with Parkinson's disease and related synucleinopathies

与帕金森病和相关突触核蛋白病相关的构象α-突触核蛋白菌株的分化和表征

• 批准号: 10460906
• 负责人: Sandra Pritzkow
• 金额: $ 51.75万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10460906
• 关键词: Adopted; Affect; Animal Model; Animals; Area; Behavioral; Biochemical; Biological; Biological Assay; Brain; Brain Injuries; Cell model; Cells; Clinical; Clinical Treatment; Dementia with Lewy Bodies; Deposition; Detection; Development; Differential Diagnosis; Disease; Early Diagnosis; FYN gene; Foundations; Goals; Histologic; In Vitro; Individual; Injections; Liquid substance; Mammalian Cell; Molecular Conformation; Morphology; Multiple System Atrophy; Neurodegenerative Disorders; Parkinson Disease; Pathogenesis; Pathogenicity; Pathologic; Patients; Periodicity; Phenotype; Plant Roots; Play; Prion Diseases; Prions; Process; Prognosis; Property; Proteins; Sampling; Seeds; Slice; Structure; Symptoms; Techniques; Technology; Testing; Tissues; Transgenic Mice; acute toxicity; alpha synuclein; brain tissue; clinical trial enrollment; effective therapy; in vivo; individual patient; laser capture microdissection; misfolded protein; mouse model; neurotoxicity; patient stratification; personalized medicine; prion-like; protein aggregation; protein misfolding; protein misfolding cyclic amplification; screening; synucleinopathy

Synucleinopathies are a diverse group of neurodegenerative diseases characterized by misfolding, aggregation and accumulation of misfolded alpha-synuclein (αSyn) which include Parkinson‘s disease (PD), multiple system atrophy (MSA) and dementia with Lewy bodies (DLB). Despite sharing the same pathological protein, different clinical and pathological phenotypes are typically observed in different synucleinopathies. One of the main obstacles to develop effective treatments for synucleinopathies is the lack of an early diagnosis before the onset of brain damage and clinical symptoms of the disease. Clinically, it is very challenging to discriminate between some of the synucleinopathies (e.g. PD and MSA), especially at early stages of the disease. This is an important problem because these diseases have different prognosis and modes of treatment. Recently, we developed the protein misfolding cyclic amplification (PMCA, also known as RT- QuIC) technology for highly sensitive and specific detection of αSyn oligomers in biological fluids of patients affected by synucleinopathies. αSyn-PMCA utilizes the prion-like seeding mechanism to cyclically amplify the process of protein misfolding, enabling the efficient amplification of small quantities of αSyn oligomers, facilitating their detection. We have evidence that the product of αSyn-PMCA coming from CSF samples of patients affected by PD and MSA can be experimentally distinguished. Thus, we hypothesize that αSyn aggregates associated with different synucleinopathies correspond to different conformational strains/ sub-strains of αSyn that can be faithfully amplified by αSyn-PMCA and the product can be differentiated by their biochemical, structural and biological properties. In this study, our main goal is to detect, differentiate and amplify minute amounts of αSyn strains/ sub-strains from biological fluids and brain tissue of patients with synucleinopathies by an in-vitro seeding/ amplification assay, and characterize these αSyn conformational aggregates using various biochemical, structural, and biological techniques. The findings obtained here will lay the foundation towards the development of a biochemical test for differential diagnosis of synucleinopathies by sensitive and specific detection of different αSyn strains/sub-strains, which will help in patients’ stratification, target enrollment for clinical trial and personalized treatment.

突触核蛋白病是一组不同的神经退行性疾病，其特征是错误折叠、 错误折叠的 α-突触核蛋白 (αSyn) 的聚集和积累，包括帕金森病 疾病（PD）、多系统萎缩（MSA）和路易体痴呆（DLB）。 共享相同的病理蛋白，不同的临床和病理表型通常是 在不同的突触核蛋白病中观察到的这一现象是开发有效治疗方法的主要障碍之一。 对于突触核蛋白病来说，在脑损伤和临床发生之前缺乏早期诊断 在临床上，区分某些疾病的症状非常具有挑战性。 突触核蛋白病（例如 PD 和 MSA），尤其是在疾病的早期阶段。 重要的问题，因为这些疾病有不同的预后和治疗方式。 最近，我们开发了蛋白质错误折叠循环扩增（PMCA，也称为RT- QuIC) 技术，用于对生物体液中的 αSyn 寡聚物进行高灵敏度和特异性检测 αSyn-PMCA 利用类似朊病毒的播种机制来治疗受突触核蛋白病影响的患者。 循环放大蛋白质错误折叠的过程，从而实现小分子的有效放大 大量的 αSyn 寡聚体，有利于它们的检测。 来自 PD 和 MSA 患者脑脊液样本的 αSyn-PMCA 可以进行实验 因此，我们追求与不同相关的αSyn聚集体。 突触核蛋白病对应于 αSyn 的不同构象菌株/亚菌株，可以 通过 αSyn-PMCA 忠实地扩增，产物可以通过其生化、 在这项研究中，我们的主要目标是检测、区分和鉴定。 从生物体液和脑组织中扩增微量的 αSyn 菌株/亚菌株 通过体外接种/扩增测定法对患有突触核蛋白病的患者进行分析，并表征这些患者 使用各种生化、结构和生物技术的 αSyn 构象聚集体。 这里获得的研究结果将为生化测试的开发奠定基础 通过灵敏且特异地检测不同的 αSyn 来鉴别诊断突触核蛋白病 菌株/亚菌株，这将有助于患者分层、临床试验的目标招募和 个性化治疗。