How Serine-129 Phosphorylation Status Affects the Spreading of α-Synuclein Pathology in Vivo: a Study in Knock-in Animals

Serine-129 磷酸化状态如何影响体内 α-突触核蛋白病理学的传播：敲入动物研究

• 批准号: 10736995
• 负责人: Ulf Dettmer
• 金额: $ 219.27万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10736995
• 关键词: Acceleration; Affect; Alabama; Alzheimer' s Disease; Animal Model; Animals; Behavior; Binding; Biochemical; Biological; Biology; Brain; Brain Diseases; Brain region; Cells; Data; Dementia with Lewy Bodies; Development; Disease; Disparate; Drug Targeting; Electrophysiology (science); Exocytosis; Genotype; Goals; Health; Hippocampus; Homeostasis; Knock-in; Knock-in Mouse; Knowledge; Lead; Lesion; Lewy Bodies; Lewy neurites; Methods; Missense Mutation; Mission; Modeling; Modification; Molecular; Multiple System Atrophy; Mus; National Institute of Neurological Disorders and Stroke; Nervous System; Neurites; Neurodegenerative Disorders; Neurons; Outcome; Parkinson Disease; Pathogenesis; Pathogenicity; Pathologic; Pathology; Pathway interactions; Patients; Phosphorylation; Phosphotransferases; Physiological; Physiology; Play; Post-Translational Protein Processing; Predisposition; Procedures; Protein Dephosphorylation; Proteins; Public Health; Recombinants; Research; Research Personnel; Rodent; Role; Serine; Slice; Surface; Synapses; Testing; Vesicle; alpha synuclein; biomarker development; drug development; early onset; extracellular; in vivo; in vivo Model; innovation; insight; knockin animal; mouse model; nervous system disorder; neural; novel; pharmacologic; pre-formed fibril; preservation; prevent; protein complex; proteostasis; synucleinopathy; trafficking; Acceleration; Affect; Alabama; Alzheimer' s Disease; Animal Model; Animals; Behavior; Binding; Biochemical; Biological; Biology; Brain; Brain Diseases; Brain region; Cells; Data; Dementia with Lewy Bodies; Development; Disease; Disparate; Drug Targeting; Electrophysiology (science); Exocytosis; Genotype; Goals; Health; Hippocampus; Homeostasis; Knock-in; Knock-in Mouse; Knowledge; Lead; Lesion; Lewy Bodies; Lewy neurites; Methods; Missense Mutation; Mission; Modeling; Modification; Molecular; Multiple System Atrophy; Mus; National Institute of Neurological Disorders and Stroke; Nervous System; Neurites; Neurodegenerative Disorders; Neurons; Outcome; Parkinson Disease; Pathogenesis; Pathogenicity; Pathologic; Pathology; Pathway interactions; Patients; Phosphorylation; Phosphotransferases; Physiological; Physiology; Play; Post-Translational Protein Processing; Predisposition; Procedures; Protein Dephosphorylation; Proteins; Public Health; Recombinants; Research; Research Personnel; Rodent; Role; Serine; Slice; Surface; Synapses; Testing; Vesicle; alpha synuclein; biomarker development; drug development; early onset; extracellular; in vivo; in vivo Model; innovation; insight; knockin animal; mouse model; nervous system disorder; neural; novel; pharmacologic; pre-formed fibril; preservation; prevent; protein complex; proteostasis; synucleinopathy; trafficking

SUMMARY/ABSTRACT Parkinson’s disease (PD), dementia with Lewy bodies (DLB), multiple-system atrophy (MSA), and certain forms of Alzheimer’s disease (AD) are ‘synucleinopathies’ - brain diseases characterized by lesions (Lewy bod- ies/Lewy neurites) rich in α-synuclein (αS). Disease-modifying treatments are not available, in part due to a lack of insight into how native αS dynamics becomes aberrant. Our long-term goal is to understand in detail how normal αS biology goes awry and to develop strategies to preserve/reestablish the normal physiological state and function of αS. αS in Lewy bodies/neurites is often phosphorylated on serine-129 (pS129), and the kinase(s) involved have been discussed as potential drug targets. However, our recent research also suggests a normal physiological role of pS129 in regulating synaptic activity (pS129 is reversibly induced by neural activity). Our overall objectives in this application are to (i) identify the effect of pS129 on the seeded intraneuronal aggregation of αS, and (ii) to determine the relevance of pS129 for the spreading of αS pathology in animal brains. Our central hypothesis is that both lack and excess of pS129 may trigger αS aggregation in the first place (because normal αS dynamics are perturbed in the absence of the “right” level of pS129). The subsequent effect on spreading is exploratory. The rationale for this project is that understanding both normal and seeded αS phosphorylation is likely to offer new insight for the development of strategies to preserve αS homeostasis, correct αS imbalance and quantify signatures of αS pathology. Taking advantage of our novel S129 phospho-deficient (S129A) and - mimicking (S129D) knock-in (KI) mouse models, we propose the following specific aims: 1) Effects of PFFs and patient-derived seed on αS aggregation in cultured WT, S129AKI, and S129DKI neurons. 2) Effects of PFFs and patient-derived seed on αS aggregation and pathogenic spread in WT, S129AKI, and S129DKI mice. Under Aim 1, primary rodent cortical neuron cultures (WT, S129AKI, and S129DKI) will be used to determine the seeding effects of PFFs and patient-derived brain extracts± synaptic activity. Synaptotoxicity mechanisms will be studied in hippocampal slices of all genotypes. In Aim 2, we will determine the effects of pS129 on αS pathogenic spread in our animal models in vivo. As one outcome, we hope to have a comprehensive view on initiation vs. spreading of pathology as a function of pS129. The proposed research is innovative because it employs novel rodent KI models of pS129 loss and pS129 excess. To our knowledge, such models have never been treated with αS “seed” before. A systemic analysis of pS129 on initiation and spreading of abnormal αS biology has not been done. Our combination of cellular, brain slice, and animal models is also innovative. Our contribution will be significant because it is expected to provide novel, paradigm-shifting insight into how normal αS biology is per- turbed in disease, and how the perturbation spreads within the brain. Corroborating that pS129 plays a key role in disease initiation and spreading, and understanding how and why, is an important step towards a comprehen- sive view of αS in health and disease with major implications for drug and biomarker development.

摘要/摘要 帕金森氏病（PD），有路易尸体的痴呆症（DLB），多系统萎缩（MSA）和某些 阿尔茨海默氏病（AD）的形式是“突触核酸病变” - 以病变为特征的脑部疾病（Lewy身体 - IES/LEWY神经运动）富含α-突触核蛋白（αs）。不可用疾病改良治疗，部分原因是缺乏 深入了解天然αs动态如何变得异常。 我们的长期目标是详细了解如何 正常α的生物学出现问题，并制定保留/重新建立正常生理状态的策略 和αs的功能。路易体/神经突中的α通常在丝氨酸-129（PS129）和激酶（S）上磷酸化。 涉及作为潜在药物靶标的讨论。但是，我们最近的研究也表明正常 PS129在调节性突触活动中的生理作用（PS129是由神经活动可逆诱导的）。我们的 本应用程序中的总体目标是（i）确定pS129对种子神经内神经元聚集的影响 αs和（ii）确定PS129与动物大脑中αS病理的相关性。我们的中心 假设是，缺乏和过量的PS129首先可能触发αs聚集（因为正常） 在没有PS129的“右”水平的情况下，αS动力学会受到干扰。随后对传播的影响是 探索性。该项目的理由是了解正常和种子的αs磷酸化是 可能提供新的见解，以制定保持αs体内平衡的策略，纠正αS失衡 并量化αs病理的特征。利用我们的新型S129磷酸化（S129A）和 - 模仿（S129D）敲入（KI）小鼠模型，我们提出以下特定目的：1）PFF和 培养的WT，S129AKI和S129DKI神经元中αs聚集的患者衍生种子。 2）PFF和 在WT，S129AKI和S129DKI小鼠中，患者衍生的种子在αs聚集和致病性传播中。在目标下 1，将使用初级啮齿动物皮质神经元培养物（WT，S129AKI和S129DKI）来确定播种 PFF和患者衍生的大脑提取物的影响±突触活动。突触毒性机制将研究 在所有基因型的海马切片中。在AIM 2中，我们将确定PS129对αs致病性差异的影响 在我们的动物模型中。作为一个结果，我们希望对启动与传播有全面的看法 病理与PS129的关系。拟议的研究具有创新性，因为IT员工新颖 PS129损耗和PS129的模型超过。据我们所知，这种模型从未用αs处理 之前的“种子”。 PS129在异常αs生物学的启动和扩散的系统分析尚未是 完毕。我们的细胞，脑切片和动物模型的结合也是创新的。我们的贡献将是 意义重大，因为它有望提供新颖的，范式转变的见解 受到疾病的影响，以及扰动如何在大脑内部传播。证实PS129起关键作用 在疾病的启动和扩散以及理解如何以及为什么是迈向完整的重要一步 对健康和疾病中αs的看法对药物和生物标志物发育产生了重大影响。