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 帕金森病 (PD)、路易体痴呆 (DLB)、多系统萎缩症 (MSA) 和某些 阿尔茨海默病 (AD) 的一种形式是“突触核蛋白病”——以病变为特征的脑部疾病（路易体 ies/路易神经突）富含 α-突触核蛋白 (αS)，因此无法获得缓解疾病的治疗，部分原因是缺乏。 深入了解天然 αS 动力学如何变得异常。 我们的长期目标是了解 αS 生物学 详细说明并制定策略以保留/重建 αS 的正常生理状态和功能。 路易体/神经突通常在丝氨酸 129 (pS129) 上被磷酸化，并且所涉及的激酶已被 然而，pS129 也可能在突触抑制方面具有正常的生理作用。 根据我们的观察，pS129 是由神经活动可逆诱导的，我们在此应用中的总体目标。 阳离子的目的是（i）确定活动调节 pS129 的突触机制，以及（ii）确定相关 pS129 在突触中发挥 αS 功能的作用 我们的中心假设是 polo 样激酶 2 (Plk2) 磷酸化。 响应突触活动，在丝氨酸 129 处标记 αS，从而微调 αS 功能。 该项目的目的是了解正常的突触 αS 磷酸化可能为发育提供新的见解 保持 αS 稳态、纠正 αS 失衡和量化 αS 病理特征的策略。 提出以下具体目标：1）确定在neu-过程中控制可逆αS pS129的机制 2) 确定可逆 pS129 在微调 αS 功能中的作用。 皮层神经元培养物将用于确认 Plk2 是在神经元活动期间介导 pS129 的激酶 并确定可能触发 pS129 的 αS 和 Plk2 分子变化 主要发现将在小鼠中得到证实。 海马切片和丰富环境的小鼠模型在第二个目标中，我们将研究其影响。 pS129 对 αS 生物学的体外和体内功能最重要的是，我们将表征 S129 磷酸化。 功能缺陷（S129A）和模拟（S129D）敲入小鼠模型，特别关注多巴胺 所提出的研究具有创新性，因为它侧重于苏尔寿实验室的既定方法。 关于 αS 正常部位（成熟神经元突触）的动态 αS S129 磷酸化，考虑突触 活性作为一个重要参数，识别关键蛋白质，解决功能后果，并扩展细胞 虽然 pS129 已被广泛研究，但之前的大多数工作都集中在其在体内的作用。 病理沉积物的贡献将是重大的，因为它有望提供新颖的范式。 转变对突触正常 αS 生物学的认识，证实 αS 修饰通常与 疾病是正常发生的，了解疾病如何发生以及为何发生，是迈向全面了解疾病的重要一步。 αS 在健康和疾病中的重要观点对药物和生物标志物的开发具有重大影响。 追求“与阿尔茨海默病相关的痴呆症的建议”中概述的研究重点 它重点关注解决 AD 相关痴呆症 (ADRD)，特别是 DLB 和 PD 痴呆症。