Investigation of TDP-43 Function and Toxicity in C. elegans

TDP-43 在秀丽隐杆线虫中的功能和毒性研究

• 批准号: 8061577
• 负责人: Christopher D. Link
• 金额: $ 32.57万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8061577
• 关键词: Alleles; Alzheimer' s Disease; Biochemical; Biological; Biological Models; Caenorhabditis elegans; Cell Culture Techniques; Cell Survival; Co-Immunoprecipitations; Complement; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Deposition; Disease; Frontotemporal Dementia; Frontotemporal Lobar Degenerations; Functional disorder; Gene Targeting; Genes; Genetic Screening; Health; Homologous Gene; Human; Huntington Disease; Immunohistochemistry; Informatin; Investigation; Lewy Body Disease; Mammalian Cell; Messenger RNA; Metabolism; Modeling; Molecular; Molecular Genetics; Movement; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Dysfunction; Neurons; Nuclear Protein; Orthologous Gene; Parkinson Disease; Pathology; Pathway interactions; Pattern; Phenotype; Pick Disease of the Brain; Play; Primary Lateral Sclerosis; Progranulin; Proteins; Publishing; RNA Interference; RNA Splicing; Reporter; Role; Screening procedure; Series; Small Interfering RNA; Stress; Synapses; Testing; Tissues; Toxic effect; Transcript; Transfection; Transgenic Organisms; Variant; Work; Writing; Yeasts; age related; amyloid peptide; base; design; effective therapy; gain of function; gene function; human Huntingtin protein; in vivo; in vivo Model; loss of function; loss of function mutation; mutant; neuron loss; neurotoxic; neurotoxicity; overexpression; polyglutamine; protein TDP-43; protein aggregate; research study; response; synuclein; trafficking; yeast two hybrid system

DESCRIPTION (provided by applicant): For many neurodegenerative diseases, both effective treatments and underlying causes are unknown. A specific protein, TDP-43, has recently been shown to be abnormally deposited in multiple neurodegenerative diseases, including Amyotropic Lateral Sclerosis (ALS) and Frontotemporal Lobar Dementia (FTLD). TDP-43 (TARDBP) is a conserved, broadly expressed nuclear protein with demonstrated roles in mRNA splicing and stability. By analogy with other neurodegenerative diseases associated with specific protein inclusions or aggregates (e.g., huntingtin in Huntington's, 1-synuclein in Parkinson's, 1-amyloid peptide in Alzheimer's, etc.), there are good reasons to believe that TDP-43 plays a causal role in neurodegenerative pathology. We have expressed human TDP-43 in C. elegans in order to assess the effects of TDP-43 overexpression in an in vivo model. Our experiments show that transgenic worms with neuronal expression of human TDP-43 have an uncoordinated movement phenotype that is indicative of neuronal dysfunction. Thus, our data support a neurotoxic role for TDP-43. The molecular mechanism(s) by which TDP-43 may be neurotoxic are unclear, in part because the full range of TDP-43 function is unknown. TDP-43-positive inclusions occur in FTLD cases caused by loss-of-function mutations in progranulin but the biological connection between TDP-43 and progranulin has not been established. In this project, we will seek to identify the conserved functions of TDP-43 (and progranulin), and the molecular and cellular mechanisms by which TDP-43 causes neurodegeneration. Specifically, we will carefully characterize C. elegans strains with deletions of TDP-43 and progranulin to determine the functions of these genes at the organismal level. We will also generate and characterize a series of transgenic C. elegans strains expressing variants of human TDP-43 designed to elucidate the molecular mechanisms of TDP-43 toxicity. These transgenic strains will also be used in forward genetic screens to identify components of the TDP-43 neurotoxic pathway. The C. elegans studies will be complemented by parallel cell culture studies, which will serve to validate and extend findings made in the C. elegans model system. PUBLIC HEALTH RELEVANCE: For many neurodegenerative diseases, both effective treatments and underlying causes are unknown. A specific protein, TDP-43, has recently been shown to be abnormally deposited in multiple neurodegenerative diseases, including Amyotropic Lateral Sclerosis (ALS) and Frontotemporal Lobar Dementia (FTLD). In this project we will use model systems to determine the functions of TDP-43 and how its deposition may cause neurodegeneration.

描述（由申请人提供）：对于许多神经退行性疾病，有效的治疗方法和根本原因尚不清楚。最近发现一种特定的蛋白质 TDP-43 在多种神经退行性疾病中异常沉积，包括肌萎缩侧索硬化症 (ALS) 和额颞叶痴呆 (FTLD)。 TDP-43 (TARDBP) 是一种保守的、广泛表达的核蛋白，已被证实在 mRNA 剪接和稳定性中发挥作用。通过与与特定蛋白质内含物或聚集体相关的其他神经退行性疾病（例如亨廷顿病中的亨廷顿蛋白、帕金森病中的 1-突触核蛋白、阿尔茨海默病中的 1-淀粉样肽等）进行类比，我们有充分的理由相信 TDP-43 在神经退行性病理学中发挥因果作用。我们在线虫中表达了人 TDP-43，以评估体内模型中 TDP-43 过表达的影响。我们的实验表明，神经元表达人 TDP-43 的转基因蠕虫具有不协调的运动表型，表明神经元功能障碍。因此，我们的数据支持 TDP-43 的神经毒性作用。 TDP-43 具有神经毒性的分子机制尚不清楚，部分原因是 TDP-43 的全部功能尚不清楚。 TDP-43 阳性包涵体发生在由颗粒体蛋白前体功能丧失突变引起的 FTLD 病例中，但 TDP-43 和颗粒体蛋白前体之间的生物学联系尚未建立。在这个项目中，我们将寻求确定 TDP-43（和颗粒体蛋白前体）的保守功能，以及 TDP-43 引起神经变性的分子和细胞机制。具体来说，我们将仔细表征删除了 TDP-43 和颗粒体蛋白前体的线虫菌株，以确定这些基因在生物水平上的功能。我们还将生成并表征一系列表达人类 TDP-43 变体的转基因线虫菌株，旨在阐明 TDP-43 毒性的分子机制。这些转基因菌株还将用于正向遗传筛选，以鉴定 TDP-43 神经毒性途径的成分。线虫研究将得到平行细胞培养研究的补充，这将有助于验证和扩展线虫模型系统中的发现。公共卫生相关性：对于许多神经退行性疾病，有效的治疗方法和根本原因尚不清楚。最近发现一种特定的蛋白质 TDP-43 在多种神经退行性疾病中异常沉积，包括肌萎缩侧索硬化症 (ALS) 和额颞叶痴呆 (FTLD)。在这个项目中，我们将使用模型系统来确定 TDP-43 的功能以及它的沉积如何导致神经变性。