Project 2

项目2

基本信息

批准号：
8249451
负责人：
MARTINA H WIEDAU-PAZOS
金额：
$ 23.12万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-08-11 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8249451
关键词：
Adolescent Adverse effects Affect Age Amyotrophic Lateral Sclerosis Animals Antibodies Area Axon Behavior Behavioral Biochemical Biological Biological Assay Biological Markers Biological Models Cell Culture System Cell Death Cell Extracts Cell Survival Cells Cellular Morphology Cessation of life Clinical Coculture Techniques Cohort Effect Collaborations Crystallins Detection Development Diagnostic Procedure Disease Disease Pathway Disease model Drug Delivery Systems Electrophysiology (science)Epitopes Evaluation Event Exhibits Fluorescence Microscopy Free Radical Formation Future Genes Goals Grant Green Fluorescent Proteins Human In Vitro Investigation Knowledge Laboratories Length Link Location Measurement Measures Mediating Membrane Potentials Metals Microtubules Mitochondria Modeling Molecular Molecular Conformation Motor Motor Neurons Mouse Strains Movement Mus Nerve Degeneration Neuroglia Neurons Organelles Pathway interactions Patients Pattern Peptides Pharmaceutical Preparations Physiological Play Preparation Process Property Proteins Publishing Reporter Research Role Signal Pathway Spinal Cord Staging Staining method Stains Stem cells Synapses Synchrotrons System Techniques Testing Tetanus Helper Peptide Therapeutic Time Tissues Toxic effect Transfection Transgenes Transgenic Organisms Ubiquitin Viral Vector Work base cell type cytochrome c design human disease human embryonic stem cell improved in vivo inhibitor/antagonist mitochondrial membrane motor neuron degeneration mouse model multicatalytic endopeptidase complex mutant neuron loss overexpression oxidation prevent programs promoter protein aggregate protein aggregation relating to nervous system research study superoxide dismutase 1 synaptogenesis uptake vector

项目摘要

Aggregation and fibrillation of SODl have been implicated in disease mechanisms of Amyotrophic Lateral Sclerosis (ALS), and it is a major new goal of this Program Project renewal to develop better biological assays to study the toxicity of these multimeric forms of SODl in systems that will be more relevant to the disease in humans. In Project 2 we further develop and use a human cell culture system that closely models important cell biological aspects of motor neuron degeneration¿our recently developed human embryonic stem cell-derived motor neuron (HESC-MN) system. The cells have distinct advantages over other model systems as they represent the major cell type that degenerates in ALS and they are fully human. The cells express identifying neuronal markers, exhibit electrophysiological function typical for mature motor neurons, and can be co-cultured with other neuronal and non-neuronal cells. Transfection of these cells to express ALS-SODl proteins causes deleterious effeds on cell survival and morphology. Importantly for this project, we have recently shown that exogenously added ALS-SODl protein multimers are taken up quite well. We will utilize these cells to study the toxicity of SODl protein multimers and aggregates at different stages of their formation and relate it to the progression of motor neuron degeneration. This research plan outlines a highly collaborative, step-by-step approach to evaluate spontaneous and induced mutant and WT SODl aggregate formation in motor neurons, followed by an investigation of the consequences of SODl aggregates on neurodegenerative mechanisms and, finally, by using pharmacological inhibitors of SODl aggregation to investigate whether reduced SODl aggregation can prevent motor neuron death.

肌萎缩性侧面的疾病机制已隐含SODL的聚集和纤颤硬化症（ALS），这是该计划项目更新的主要新目标，可以开发出更好的生物学测定法研究这些多个例子SODL在系统中与该疾病更相关的系统的毒性人类。在项目2中，我们进一步开发并使用了密切建模重要细胞的人类细胞培养系统运动神经元变性的生物学方面。我们最近开发的人类干细胞衍生运动神经元（HESC-MN）系统。这些单元在代表其他模型系统中具有明显的优势在ALS中退化的主要细胞类型，它们是完全人类的。细胞表达识别神经元的细胞标记物，典型的成熟运动神经元的暴露电生理功能，可以与其他神经元和非神经元细胞。转染这些细胞以表达ALS-SODL蛋白会引起有害细胞的生存和形态。重要的是，对于这个项目，我们最近表明外源添加的ALS-SODL蛋白多聚体非常好。我们将利用这些细胞研究 SODL蛋白多聚体和聚集体在其形成的不同阶段的毒性，并将其与运动神经元变性的进展。该研究计划概述了一个高度协作的，逐步的评估运动神经元中的赞助和诱导突变体和WT SODL骨料形成的方法，然后是SODL骨料对神经退行性机制的后果的投资，以及最后，通过使用SODL聚集的药物抑制剂来研究SODL是否减少聚集可以预防运动神经元死亡。