Elucidating the biological differences between distinct fibrillar and non-fibrillar alpha-synuclein inclusions in human stem-cell models

阐明人类干细胞模型中不同纤维状和非纤维状 α-突触核蛋白内含物之间的生物学差异

• 批准号: 10739667
• 负责人: Vikram Khurana
• 金额: $ 15.65万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10739667
• 关键词: ARHGEF1 gene; Actins; Address; Affect; Alzheimer like pathology; Alzheimer' s Disease; Alzheimer' s disease pathology; Amyloid; Autopsy; Biological; Biological Process; Biology; Brain; Brain Pathology; CRISPR screen; CRISPR/Cas technology; Cell model; Cell physiology; Cells; Chronic; Cytopathology; Cytoskeleton; Defect; Disease; Exhibits; F-Actin; Functional disorder; Genes; Genetic; Genetic Screening; Genomics; Grant; Human; Induced pluripotent stem cell derived neurons; Investigation; Lead; Lewy Bodies; Lewy Body Disease; Lewy body pathology; Link; Literature; MAPT gene; Mediating; Membrane; Microfilaments; Modeling; Molecular; Mutation; Neurodegenerative Disorders; Neuroglia; Neurons; Organelles; Parents; Parkinson Disease; Pathologic; Pathology; Pathway interactions; Patients; Persons; Phenotype; Polymers; Proteins; RHOA gene; Regulation; Reporting; Rod; Synaptic Vesicles; Tauopathies; Testing; Toxic effect; Validation; Vesicle; alpha synuclein; cell type; cytotoxicity; genome-wide; grasp; human stem cells; link protein; mitochondrial dysfunction; neurotoxicity; new therapeutic target; novel; overexpression; pharmacologic; polymerization; protein aggregation; protein transport; stem cell model; synucleinopathy; targeted exome sequencing; tau Proteins; tau aggregation; trafficking

Project Summary Aggregation of specific proteins within neurons and glia comprise the hallmark pathologies of neurodegenerative diseases (ND) like Alzheimer’s (AD) and Parkinson’s disease (PD). In AD, the aggregating protein is b-amyloid (Ab), and in PD it is a-synuclein (a-syn). Intriguingly, these two pathologies often coexist and the significance of this is unknown. Many cellular defects are detected in the presence of a-syn mutations or overexpression, prominently including defective vesicle trafficking. Outstanding questions remain—how does a-syn impart toxicity, and how does it exert effects on a wide span of cellular pathways? How does stage of the pathology, including the formation of distinct subtypes of a-syn inclusions, alter the vulnerability of the cell? What is the connection between a-syn and the AD-linked protein Ab that is often seen to coexist in the brains of patients with synucleinopathies? We previously conducted a genome-wide CRISPR/Cas9 screen for genetic modifiers of a-syn toxicity in a human cellular model that captures advanced membrane-rich a-syn aggregates highly reminiscent of human postmortem brain pathology in PD. We also conducted a targeted exome sequencing screen in synucleinopathy patients, focusing on known modulators of both a-syn and Ab cytotoxicity. Interestingly, top hits from both of these approaches converged on genes related to actin cytoskeleton regulation. The convergence of hits from these two screens led us to hypothesize that a key aspect of a-syn toxicity relates to altered actin cytoskeletal stabilization and that this may be a key point of convergence of both a-syn and Ab cellular toxicity. Indeed, the actin cytoskeleton orchestrates organization of cellular organelles and substructures and others have postulated its dysregulation may play a role in many of the a-syn- and Ab-mediated cellular defects. Our genetic investigations in human cells and patients now pinpoint a specific set of genes that might mediate this crosstalk between AD and PD pathologies. In this proposal, we aim to (1) examine whether the actin cytoskeleton contributes to toxicity in synucleinopathy iPSC-derived neuron models, (2) conduct a pooled secondary screen in iPSC-derived neuron models and validate genetic modifiers of a-syn toxicity, and (3) examine whether a-syn-mediated actin cytoskeleton defects are modulated by Ab to understand the convergence between a-syn and Ab toxicity. Understanding the molecular and genetic underpinnings of PD, and any possible connection with AD, will ultimately contribute to a better grasp of the disease and help uncover novel therapeutic targets.

项目概要 神经元和神经胶质细胞内特定蛋白质的聚集，构成神经退行性疾病的标志性病理学 阿尔茨海默病 (AD) 和帕金森病 (PD) 等疾病 (ND) 在 AD 中，聚集蛋白是 b-淀粉样蛋白。 (Ab)，而在 PD 中，它是 a-突触核蛋白 (a-syn) 有趣的是，这两种病理学经常共存，并且具有重要意义。 这是未知的，许多细胞缺陷是在 a-syn 突变或过度表达的情况下检测到的。 突出地包括有缺陷的囊泡运输，但仍然存在悬而未决的问题——a-syn 是如何传递的。 毒性，以及它如何对广泛的细胞途径产生影响？ 包括形成不同亚型的 a-syn 内含物，改变细胞的脆弱性是什么？ a-syn 和 AD 相关蛋白 Ab 之间的联系，通常在患者大脑中共存 与突触核蛋白病？我们之前进行了全基因组 CRISPR/Cas9 筛选，以寻找 人类细胞模型中的 a-syn 毒性，捕获先进的富含膜的 a-syn 聚集体 我们还进行了针对性的外显子组测序。 对突触核蛋白病患者进行筛选，重点关注已知的 a-syn 和 Ab 细胞毒性调节剂。 模棱两可，这两种方法的热门结果都集中在与肌动蛋白细胞骨架调节相关的基因上。 这两个屏幕的命中率的融合使我们认识到 a-syn 毒性的一个关键方面与 改变肌动蛋白细胞骨架稳定性，这可能是 a-syn 和 Ab 收敛的关键点 事实上，肌动蛋白细胞骨架协调细胞器和亚结构的组织。 和其他人假设其失调可能在许多 a-syn 和 Ab 介导的细胞中发挥作用 我们对人类细胞和患者的基因研究现在查明了一组可能存在缺陷的特定基因。 调解 AD 和 PD 病理之间的这种串扰在本提案中，我们的目标是 (1) 检查是否 肌动蛋白细胞骨架导致突触核蛋白病 iPSC 衍生神经元模型的毒性，(2) 进行汇总 在 iPSC 衍生的神经元模型中进行二次筛选并验证 a-syn 毒性的遗传修饰剂，以及 (3) 检查 a-syn 介导的肌动蛋白细胞骨架缺陷是否受 Ab 调节，以了解 a-syn 和 Ab 毒性之间的趋同了解 PD 的分子和遗传基础，以及 任何与 AD 可能存在的联系，最终都将有助于更好地了解这种疾病并帮助发现 新的治疗靶点。

项目成果

Effects of Excess Brain-Derived Human α-Synuclein on Synaptic Vesicle Trafficking.

• DOI: 10.3389/fnins.2021.639414
• 发表时间: 2021
• 期刊: Frontiers in neuroscience
• 影响因子: 4.3
• 作者: Román-Vendrell C;Medeiros AT;Sanderson JB;Jiang H;Bartels T;Morgan JR
• 通讯作者: Morgan JR

α-Synuclein serine129 phosphorylation - the physiology of pathology.

• DOI: 10.1186/s13024-023-00680-x
• 发表时间: 2023-11-13
• 期刊: MOLECULAR NEURODEGENERATION
• 影响因子: 15.1
• 作者: Ramalingam, Nagendran;Dettmer, Ulf
• 通讯作者: Dettmer, Ulf

The Parkinson's disease protein alpha-synuclein is a modulator of processing bodies and mRNA stability.

• DOI: 10.1016/j.cell.2022.05.008
• 发表时间: 2022-06-09
• 期刊: CELL
• 影响因子: 64.5
• 作者: Hallacli, Erinc;Kayatekin, Can;Nazeen, Sumaiya;Wang, Xiou H.;Sheinkopf, Zoe;Sathyakumar, Shubhangi;Sarkar, Souvarish;Jiang, Xin;Dong, Xianjun;Di Maio, Roberto;Wang, Wen;Keeney, Matthew T.;Felsky, Daniel;Sandoe, Jackson;Vahdatshoar, Aazam;Udeshi, Namrata D.;Mani, D. R.;Carr, Steven A.;Lindquist, Susan;De Jager, Philip L.;Bartel, David P.;Myers, Chad L.;Greenamyre, J. Timothy;Feany, Mel B.;Sunyaev, Shamil R.;Chung, Chee Yeun;Khurana, Vikram
• 通讯作者: Khurana, Vikram

Brain region-specific susceptibility of Lewy body pathology in synucleinopathies is governed by α-synuclein conformations.

• DOI: 10.1007/s00401-022-02406-7
• 发表时间: 2022-04
• 期刊: Acta neuropathologica
• 影响因子: 12.7
• 作者: de Boni L;Watson AH;Zaccagnini L;Wallis A;Zhelcheska K;Kim N;Sanderson J;Jiang H;Martin E;Cantlon A;Rovere M;Liu L;Sylvester M;Lashley T;Dettmer U;Jaunmuktane Z;Bartels T
• 通讯作者: Bartels T

Adding hydrophobicity or positive charges to the cytosolic half of the α-synuclein 3-11 helix increases membrane association and S129 phosphorylation.

添加疏水性或正电荷到 α-突触核蛋白 3-11 螺旋的胞质一半会增加膜结合和 S129 磷酸化。

• DOI: 10.1002/1873-3468.14773
• 发表时间: 2024
• 期刊: FEBS letters
• 影响因子: 3.5
• 作者: Shimanaka,Kazuma;Shi,Bryan;Brontesi,Lisa;Alnakhala,Heba;Jayanthi,Vidyashree;Subramanian,Kanagaraj;Ramalingam,Nagendran;Tripathi,Arati;Dettmer,Ulf
• 通讯作者: Dettmer,Ulf