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

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.

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