Molecular Mechanisms of SOD1-linked ALS (P01)

描述（由申请人提供）：该计划项目汇集了五名具有非常不同的专业知识和实验能力的研究人员，共同致力于阐明S 0 D1相关家族性肌萎缩侧索硬化症（fALS）发病机制的潜在机制。已经确定S 0 D1多聚体和较大的聚集体与疾病相关，但毒性物质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S 0 D1及其聚集相关的生化特性，揭示疾病起始和进展的线索，利用这种理解开发多聚化的靶向阻断剂。PPG合作将包括五名主要研究人员，四个项目和一个技术核心。项目1（Joan Valentine博士）和项目4（大卫艾森伯格博士）将采用体外结构和生物物理方法研究S 0 D1多聚化的机制和多聚体的结构，目的是了解多聚化的机制和设计聚集抑制剂。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，项目3（大卫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S 0 D1的变化。后一个项目的特别重点将是体内金属负载，因为它涉及S 0 D1的稳定性，和金属稳态，因为它涉及细胞毒性。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组织将被用作验证研究结果和测试新假设的来源。

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.