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

基本信息

批准号：
8453483
负责人：
Christopher D. Link
金额：
$ 31.43万
依托单位：
UNIVERSITY OF COLORADO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-01 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8453483
关键词：
Alleles Alzheimer&apos 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 Expression Profile Gene Targeting Genes Genetic Screening Health Homologous Gene Human Huntington Disease Immunohistochemistry Informatin Investigation Lewy Body Dementia 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 Phenotype Pick Disease of the Brain Play Primary Lateral Sclerosis Progranulin Proteins Publishing RNA Interference RNA Splicing Reporter Role 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 screening synuclein trafficking yeast two hybrid system

项目摘要

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.

描述（由申请人提供）：对于许多神经退行性疾病，有效的治疗和潜在原因尚不清楚。最近已证明一种特异性蛋白TDP-43在多种神经退行性疾病中被证明异常沉积，包括肌动肌外侧硬化症（ALS）和额颞叶痴呆症（FTLD）。 TDP-43（TARDBP）是一种保守的，广泛表达的核蛋白，在mRNA剪接和稳定性中表现出作用。通过与其他与特定蛋白夹杂物或聚集体相关的神经退行性疾病（例如，亨廷顿的亨廷顿蛋白，帕金森氏症中的1-核蛋白，阿尔茨海默氏症中的1-淀粉样肽等），有充分的理由相信TDP-43有很多理由，在神经退行性病理中的作用。我们已经在秀丽隐杆线虫中表达了人类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，人类TDP-43神经元表达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可能是神经毒性的分子机制尚不清楚，部分原因是TDP-43功能的全范围尚不清楚。 TDP-43阳性夹杂物发生在由尿素损失突变引起的FTLD病例中，但尚未建立TDP-43和Progranulin之间的生物学联系。在这个项目中，我们将寻求确定TDP-43（和progranulin）的保守功能，以及TDP-43引起神经变性的分子和细胞机制。具体而言，我们将仔细地表征秀丽隐杆线虫菌株，其缺失TDP-43和Progranulin，以确定这些基因在生物水平上的功能。我们还将生成和表征一系列的转基因秀丽隐杆线虫菌株，表达人类TDP-43的变体，旨在阐明TDP-43毒性的分子机制。这些转基因菌株也将用于正向遗传筛选中，以鉴定TDP-43神经毒性途径的成分。秀丽隐杆线虫的研究将由平行细胞培养研究补充，该研究将有助于验证和扩展秀丽隐杆线虫模型系统中的发现。