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.

总结/摘要 帕金森病（PD）、路易体痴呆（DLB）、多系统萎缩（MSA）和某些 阿尔茨海默氏病（AD）的形式是"突触核蛋白病"--以病变为特征的脑疾病（Lewybod.com）。 lewy神经突）富含α-突触核蛋白（α S）。没有改善疾病的治疗方法，部分原因是缺乏 了解原生α S动力学如何变得异常。 我们的长期目标是了解α S生物学， 详细描述并制定策略以保持/重建α S的正常生理状态和功能。α S in 路易体/神经突通常在丝氨酸-129（pS129）上磷酸化，并且所涉及的激酶已经被发现。 被认为是潜在的药物靶点然而，pS129也可能在突触上具有正常的生理作用。 我们观察到pS129是由神经活动可逆诱导的。我们在这一应用中的总体目标- 阳离子是（i）确定活动调节pS129的突触机制，和（ii）确定相关的 pS129对α S功能的万斯作用。我们的中心假设是polo样激酶2（Plk 2）磷酸化- 响应突触活动，使α S在丝氨酸-129处发生磷酸化，从而微调α S功能。这样做的理由 了解正常的突触α S磷酸化可能会为开发 保护α S稳态、纠正α S失衡和量化α S病理特征的策略。我们 提出了以下具体目标：1）确定在neu过程中控制可逆α S pS129的机制， ronal活动2)确定可逆pS129在微调α S功能中的作用。在第一个目标下，初级啮齿动物 皮质神经元培养物将用于确认Plk2作为在神经元活动期间介导pS129的激酶 并鉴定可能触发pS129的α S和Plk2的分子变化。将在小鼠中证实关键发现 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 pS129对α S生物学功能的影响。最重要的是，我们将表征S129磷酸- 功能性缺陷（S129A）和模拟（S129D）敲入小鼠模型，特别关注多巴胺 使用苏尔寿实验室的既定方法进行放行。这项研究是创新的，因为它关注的是 在α S正常位置（成熟神经元的突触）的动态α S S129磷酸化，认为突触 活性作为一个重要的参数，确定关键蛋白质，解决功能的后果，并扩展细胞 培养结果在体内。尽管pS129已被广泛研究，但大多数先前的工作集中在其在以下方面的作用： 病理性沉积贡献将是重大的，因为它预计将提供新颖的，范式- 改变了对突触处正常α S生物学的认识。证实了α S修饰通常与- 与疾病发生正常，并了解如何和为什么，是一个重要的一步，以消除母鸡- α S在健康和疾病中的作用，对药物和生物标志物的开发具有重要意义。这项工作 追求在"阿尔茨海默病相关痴呆症的建议，赋予- 恩塞"。它侧重于解决AD相关痴呆（ADRD），特别是DLB和PD痴呆的优先事项。