Molecular Mechanisms of SOD1-linked ALS (P01)

SOD1 相关 ALS 的分子机制 (P01)

• 批准号: 8644318
• 负责人: JOAN Selverstone VALENTINE
• 金额: $ 95.03万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-11 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8644318
• 关键词: Address; Amyloid Fibrils; Amyotrophic Lateral Sclerosis; Animal Disease Models; Animal Model; Award; Biochemical; Biological Assay; Biological Models; Cell Culture Techniques; Cessation of life; Collaborations; Data; Disease; Disease Progression; Disease model; Disulfides; Etiology; Familial Amyotrophic Lateral Sclerosis; Gene Mutation; Goals; Homeostasis; Human; In Vitro; Investigation; Lead; Link; Metals; Methods; Modeling; Molecular; Molecular Weight; Motor Neurons; Mus; Mutation; Neurodegenerative Disorders; Neuroglia; Pathogenesis; Patients; Preparation; Principal Investigator; Process; Property; Proteins; Research Personnel; Role; Source; Spinal Cord; Stem cells; Structure; Study models; Symptoms; Testing; Tissues; Toxic effect; Vertebral column; Work; animal tissue; biophysical techniques; copper zinc superoxide dismutase; design; efficacy testing; human tissue; in vivo; inhibitor/antagonist; instrumentation; mutant; programs; public health relevance

DESCRIPTION (provided by applicant): This Program Project brings together five researchers with very different expertise and experimental capabilities to work together on elucidating the underlying mechanisms of S0D1-linked familial amyotrophic lateral sclerosis (fALS) pathogenesis. It is well established that S0D1 multimers and larger aggregates are associated with disease but the toxic species and in vivo mechanism remain unknown. The overall goals of this proposal are to gain an extensive understanding of the role of aggregation in disease, to characterize further the biochemical properties associated with mutant S0D1 and its aggregation, to uncover clues about the initiation and progression of disease, to exploit this understanding to develop targeted blockers of multimerization. The PPG collaboration will encompass five primary investigators with four projects and a technical Core. Projects 1 (Dr. Joan Valentine) and 4 (Dr. David Eisenberg) will take an in vitro structural and biophysical approach to studying the mechanism of S0D1 multimerization and the structures of the multimers, with the goals of understanding the mechanism(s) of multimerization and designing inhibitors of aggregation. Project 2 (Dr. Martina Wiedau-Pazos) will use stem cell-derived motor neurons and glia and project 3 (Dr. David Borchelt) will use a mouse,and cell culture models to probe the toxicity of multimers and to characterize the changes in mutant S0D1 that lead toward disease. A particular emphasis on the latter project will be toward in vivo metal loading as it pertains to S0D1 stability, and metal homeostasis as it pertains toward cellular toxicity. Core A (Dr. Julian Whitelegge) will serve as the backbone of these investigations by providing and maintaining the necessary instrumentation and data delivery. Disease models from projects 2 and 3 will be used to test the efficacy of inhibitors from projects 1 and 4. Finally, ALS tissue will be used as a source to validate the findings and test new hypotheses.

描述（由申请人提供）：该计划项目汇集了五位具有截然不同专业知识和实验能力的研究人员，共同努力阐明 S0D1 相关的家族性肌萎缩侧索硬化症 (fALS) 发病机制的潜在机制。众所周知，S0D1 多聚体和较大的聚集体与疾病相关，但毒性种类和体内机制仍然未知。该提案的总体目标是广泛了解聚集在疾病中的作用，进一步表征与突变体 S0D1 及其聚集相关的生化特性，揭示疾病发生和进展的线索，利用这种理解来开发多聚化的靶向阻断剂。 PPG 合作将包括五个主要研究人员、四个项目和一个技术核心。项目1（Joan Valentine博士）和4（David Eisenberg博士）将采用体外结构和生物物理方法来研究S0D1多聚化机制和多聚体结构，目的是了解多聚化机制并设计聚集抑制剂。项目 2（Martina Wiedau-Pazos 博士）将使用干细胞衍生的运动神经元和神经胶质细胞，项目 3（David Borchelt 博士）将使用小鼠和细胞培养模型来探测多聚体的毒性，并表征导致疾病的突变型 S0D1 的变化。后一个项目的特别重点是体内金属负载，因为它与 S0D1 稳定性有关，以及金属稳态，因为它与细胞毒性有关。核心 A（Julian Whitelegge 博士）将作为这些调查的骨干，提供和维护必要的仪器和数据传输。项目 2 和 3 的疾病模型将用于测试项目 1 和 4 的抑制剂的功效。最后，ALS 组织将用作验证研究结果和测试新假设的来源。

项目成果

An analysis of interactions between fluorescently-tagged mutant and wild-type SOD1 in intracellular inclusions.

细胞内包涵体中荧光标记突变体和野生型 SOD1 之间相互作用的分析。

• DOI: 10.1371/journal.pone.0083981
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Qualls,DavidA;Crosby,Keith;Brown,Hilda;Borchelt,DavidR
• 通讯作者: Borchelt,DavidR

Unbiased screen reveals ubiquilin-1 and -2 highly associated with huntingtin inclusions.

• DOI: 10.1016/j.brainres.2013.06.006
• 发表时间: 2013-08-02
• 期刊: Brain research
• 影响因子: 2.9
• 作者: Rutherford NJ;Lewis J;Clippinger AK;Thomas MA;Adamson J;Cruz PE;Cannon A;Xu G;Golde TE;Shaw G;Borchelt DR;Giasson BI
• 通讯作者: Giasson BI

Role of disulfide cross-linking of mutant SOD1 in the formation of inclusion-body-like structures.

突变体 SOD1 的二硫键交联在包涵体样结构形成中的作用。

• DOI: 10.1371/journal.pone.0047838
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Roberts,BrittanyLT;Patel,Kinaree;Brown,HildaH;Borchelt,DavidR
• 通讯作者: Borchelt,DavidR

Substantially elevating the levels of αB-crystallin in spinal motor neurons of mutant SOD1 mice does not significantly delay paralysis or attenuate mutant protein aggregation.

显着提高突变型 SOD1 小鼠脊髓运动神经元中 αB-晶状体蛋白的水平不会显着延迟麻痹或减弱突变蛋白聚集。

• DOI: 10.1111/jnc.13022
• 发表时间: 2015
• 期刊: Journal of neurochemistry
• 影响因子: 4.7
• 作者: Xu,Guilian;Fromholt,Susan;Ayers,JacobI;Brown,Hilda;Siemienski,Zoe;Crosby,KeithW;Mayer,ChristopherA;Janus,Christopher;Borchelt,DavidR
• 通讯作者: Borchelt,DavidR

A Novel SOD1 Intermediate Oligomer, Role of Free Thiols and Disulfide Exchange.

• DOI: 10.3389/fnins.2020.619279
• 发表时间: 2020
• 期刊: Frontiers in neuroscience
• 影响因子: 4.3
• 作者: Koo BK;Munroe W;Gralla EB;Valentine JS;Whitelegge JP
• 通讯作者: Whitelegge JP