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.

项目摘要 神经元和神经胶质内特定蛋白质的聚集包括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）。在AD中，聚集蛋白是b-淀粉样蛋白 (Ab)在PD中，它是α-突触核蛋白（α-syn）。有趣的是，这两种病理往往共存， 这是未知的。许多细胞缺陷是在存在α-syn突变或过表达的情况下检测到的， 突出地包括有缺陷的囊泡运输。悬而未决的问题--a-syn如何传授 毒性，以及它如何对广泛的细胞途径产生影响？病理学阶段， 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，通常被认为共存于患者的大脑中 共核蛋白病吗我们之前进行了一项全基因组CRISPR/Cas9筛选，以寻找遗传修饰剂， 高度捕获高级富膜a-syn聚集体的人细胞模型中的a-syn毒性 让人联想到帕金森病中的人类死后大脑病理学。我们还进行了靶向外显子组测序 在突触核蛋白病患者中进行筛选，重点是已知的α-syn和Ab细胞毒性的调节剂。 有趣的是，这两种方法的最高命中率都集中在与肌动蛋白细胞骨架调控相关的基因上。 这两个筛选结果的一致性使我们假设α-syn毒性的一个关键方面与 改变肌动蛋白细胞骨架的稳定性，这可能是α-syn和Ab融合的关键点。 细胞毒性事实上，肌动蛋白细胞骨架协调细胞器和亚结构的组织 其他人推测其失调可能在许多α-syn和Ab介导的细胞凋亡中起作用。 缺陷我们对人类细胞和患者的基因研究现在确定了一组特定的基因， 介导AD和PD病理之间的这种串扰。在这项建议中，我们的目的是（1）研究 肌动蛋白细胞骨架有助于突触核蛋白病iPSC衍生的神经元模型的毒性，（2）进行合并的 在iPSC衍生的神经元模型中进行二次筛选并验证α-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到Ab的调节，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， 任何可能与AD的联系，最终将有助于更好地掌握疾病，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

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

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

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

Pathogenic Mechanisms of Cytosolic and Membrane-Enriched α-Synuclein Converge on Fatty Acid Homeostasis.

细胞质和膜富集的 α-突触核蛋白的致病机制汇聚于脂肪酸稳态。

• DOI: 10.1523/jneurosci.1881-21.2022
• 发表时间: 2022
• 期刊: The Journal of neuroscience : the official journal of the Society for Neuroscience
• 作者: Tripathi,Arati;Alnakhala,Heba;Terry-Kantor,Elizabeth;Newman,Andrew;Liu,Lei;Imberdis,Thibaut;Fanning,Saranna;Nuber,Silke;Ramalingam,Nagendran;Selkoe,Dennis;Dettmer,Ulf
• 通讯作者: Dettmer,Ulf