Polo-like-kinase-2-dependent α-Synuclein Serine-129 Phosphorylation: a Physiological RoleDuring Synaptic Activity

Polo 样激酶 2 依赖性 α-突触核蛋白丝氨酸 129 磷酸化：突触活动期间的生理作用

• 批准号: 10522495
• 负责人: Ulf Dettmer
• 金额: $ 212.19万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10522495
• 关键词: Address; Alzheimer' s Disease; Alzheimer' s disease related dementia; Behavior; Binding; Biochemical; Biological; Biology; Brain; Brain Diseases; Calcineurin; Calcium; Cell Culture Techniques; Cells; Data; Dementia with Lewy Bodies; Deposition; Development; Disease; Dopamine; Drug Targeting; Electrophysiology (science); Exocytosis; Feedback; Goals; Health; Hippocampus (Brain); Homeostasis; In Vitro; Knock-in Mouse; Knowledge; Lead; Lesion; Lewy Bodies; Lewy neurites; Locales; Mediating; Methods; Mind; Missense Mutation; Mission; Modification; Molecular; Molecular Profiling; Multiple System Atrophy; Mus; National Institute of Neurological Disorders and Stroke; Neurites; Neurodegenerative Disorders; Neurons; Outcome; PLK1 gene; Parkinson Disease; Parkinson' s Dementia; Pathogenesis; Pathologic; Pathology; Pathway interactions; Pharmacology; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Physiology; Play; Post-Translational Protein Processing; Protein Phosphatase 2A Regulatory Subunit PR53; Proteins; Public Health; Recommendation; Reporting; Research; Research Priority; Rodent; Role; Serine; Slice; Sodium Channel Blockers; Surface; Synapses; Tetrodotoxin; Toxic effect; Vesicle; Work; alpha synuclein; alpha synuclein gene; base; biomarker development; calcineurin phosphatase; drug development; early onset; environmental enrichment for laboratory animals; in vivo; innovation; insight; mouse model; nervous system disorder; neurotransmitter release; novel; preservation; prevent; protein complex; proteostasis; relating to nervous system; response; symposium; synuclein; 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 αS biology in detail 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, pS129 may also have normal physiological role at synapses sup- ported by our observation that pS129 is reversibly induced by neural activity. Our overall objectives in this appli- cation are to (i) identify the synaptic mechanisms by which activity regulates pS129, and (ii) determine the rele- vance of pS129 for αS function at the synapse. Our central hypothesis is that polo-like kinase 2 (Plk2) phosphor- ylates αS at serine-129 in response to synaptic activity, thereby fine-tuning αS function. The rationale for this project is that understanding normal synaptic 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. We propose the following specific aims: 1) Identify the mechanism(s) that govern reversible αS pS129 during neu- ronal activity. 2) Identify the role of reversible pS129 in fine-tuning αS function. Under the first aim, primary rodent cortical neuron cultures will be used to confirm Plk2 as the kinase that mediates pS129 during neuronal activity and to identify molecular changes in αS and Plk2 that may trigger pS129. Key findings will be confirmed in mouse hippocampal slices and in a mouse model of enriched environment. In the second aim, we will study the effects of pS129 on αS biology functionally in vitro and in vivo. Most importantly, we will characterize S129 phospho- deficient (S129A) and -mimicking (S129D) knock-in mouse models functionally with a special focus on dopamine release, using established methods in the Sulzer lab. The proposed research is innovative, because it focuses on dynamic αS S129 phosphorylation at αS’s normal locale (the synapse of mature neurons), considers synaptic activity as an important parameter, identifies key proteins, addresses functional consequences, and extends cell culture findings to in vivo. Whereas pS129 has been widely studied, most previous work focused on its role in pathological deposits. The contribution will be significant because it is expected to provide novel, paradigm- shifting insight into normal αS biology at the synapse. Corroborating that an αS modification commonly associ- ated with disease occurs normally, 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. This work pursues research priorities outlined in “Recommendations of the Alzheimer's disease-related dementias confer- ence”. It focuses on priorities that address AD-related dementias (ADRD), specifically DLB and PD dementia.

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。 我们的长期目标是了解αs生物学 细节并制定策略来保留/重新建立αs的正常身体状态和功能。 αsin 路易体/神经突通常在丝氨酸-129（PS129）上磷酸化，涉及的激酶已经是 被视为潜在的药物靶标。但是，PS129在突触中也可能具有正常的身体作用。 通过我们的观察，即PS129是由神经活动可逆诱导的。我们在此应用中的总体目标 阳离子是（i）确定活性调节PS129的合成机制，（ii）确定相关 PS129的VANCE用于突触时αs功能。我们的中心假设是类似类球激酶2（PLK2）磷酸 - 丝氨酸129处的YLATESαs响应突触活性，从而微调αs功能。理由 项目是了解正常的突触αS磷酸化可能为开发提供新的见解 保留αs稳态，纠正αS失衡和量化αs病理学的特征的策略。我们 建议以下具体目的：1）确定在neu-期间控制可逆αsps129的机制 罗纳尔活动。 2）确定可逆PS129在微调αS函数中的作用。在第一个目标下，主要啮齿动物 皮质神经元培养物将用于确认PLK2为在神经元活性过程中介导PS129的激酶 并确定可能触发PS129的α和PLK2中的分子变化。关键发现将在鼠标中确认 海马切片和富集环境的小鼠模型。在第二个目标中，我们将研究效果 在αs生物学上的PS129在体外和体内的生物学上。最重要的是，我们将表征S129磷酸 缺乏（S129A）和-Mimicking（S129D）在功能上敲入鼠标模型，特别关注多巴胺 使用Sulzer Lab中的既定方法释放。拟议的研究具有创新性，因为它专注于 在αS正常位置（成熟神经元的突触）处的动态αSS129磷酸化上，考虑突触 活动作为重要参数，识别关键蛋白质，解决功能后果并扩展细胞 体内的培养结果。尽管PS129已被广泛研究，但大多数以前的工作都集中在其在 该贡献将是重要的，因为它有望提供新颖的范式 - 转移对突触时正常αs生物学的洞察力。证实αs修饰通常是关联的 疾病正常发生，并了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。这项工作 在“阿尔茨海默氏病有关的痴呆症会议的建议中概述的研究优先事项 - 它的重点是针对广告相关痴呆症（ADRD），特别是DLB和PD痴呆症的优先级。