Phage Display Investigations of TDP-43

TDP-43 的噬菌体展示研究

• 批准号: 7941728
• 负责人: BRIAN KENNETH KAY
• 金额: $ 19.3万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7941728
• 关键词: Amyotrophic Lateral Sclerosis; Antibodies; Bacteria; Binding; Binding Proteins; Cell Nucleus; Cells; Cytoplasmic Inclusion; DNA-Binding Proteins; Disease; Epitopes; Exhibits; Familial Amyotrophic Lateral Sclerosis; Frontotemporal Dementia; Genes; Immunoglobulin Variable Region; Inherited; Intraventricular; Investigation; Lead; Libraries; Ligands; Location; Monoclonal Antibodies; Motor Neurons; Mutate; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Nuclear; Parkinson Disease; Pathogenesis; Patients; Peptides; Phage Display; Play; Process; Proteins; Role; Specificity; Therapeutic; Toxic effect; Transgenic Mice; Ubiquitin; Work; cell type; effective therapy; frontotemporal lobar dementia-amyotrophic lateral sclerosis; insight; mutant; protein TDP-43; protein aggregate; protein function; protein misfolding; public health relevance; tau Proteins

DESCRIPTION (provided by applicant): The focus of this application is on abnormalities of TDP-43, a protein that underlies a number of neurodegenerative diseases (frontotemporal lobar dementia [FTLD], Parkinson's disease [PD], and amyotrophic lateral sclerosis [ALS]). In 2006, ubiquinated cytoplasmic inclusions in neurons in FTLD were found to contain abnormally phosphorylated fragments of aggregated TDP-43, a DNA-binding protein normally nuclear in location. Remarkably, TDP-43 inclusions are seen in all patients with sporadic ALS, the majority of patients with familial ALS (FALS), and in all patients with sporadic and familial FTLD with ubiquitin-positive, tau-negative inclusions with or without ALS. TDP-43 mutations have recently been identified in FALS patients, indicating the clear direct involvement of mutant (MT) TDP-43 in ALS. The mechanism underlying the toxicity of TDP-43 remains unclear; however, the presence of cytoplasmic aggregates suggests that misfolding of this protein is important in the pathogenesis of ALS. These aggregates may sequester TDP-43 binding-proteins important to the viability of the motor neuron (MN). In this application we plan to use phage display libraries to: (1) identify and characterize peptides that bind TDP- 43 and predict the cellular interacting proteins; (2) identify and characterize single chain fragments of variable region antibodies (scFvs) that can be used to clarify the function of TDP-43 and may have therapeutic potential. PUBLIC HEALTH RELEVANCE: Amyotrophic lateral sclerosis (ALS) is a desperate disease in which there is no cure and no effective treatment. This application involves identifying peptide ligands and generating antibodies that bind a protein that appears to be key to the pathogenesis of ALS as well as other neurodegenerative processes. The peptide ligands may be used to predict the cellular interacting proteins and the antibodies may clarify the function of the protein. Together, this work may provide insight into why mutations of this protein cause ALS, and potentially provide a direction for treatment of sporadic and inherited ALS (as well as other neurodegenerative diseases).

描述（由申请人提供）：本申请的重点是 TDP-43 的异常，TDP-43 是一种导致多种神经退行性疾病（额颞叶痴呆 [FTLD]、帕金森病 [PD] 和肌萎缩侧索硬化症 [ALS]）的蛋白质）。 2006 年，人们发现 FTLD 神经元中泛素化的细胞质内含物含有异常磷酸化的聚集 TDP-43 片段，TDP-43 是一种 DNA 结合蛋白，通常位于细胞核内。值得注意的是，TDP-43 包涵体可见于所有散发性 ALS 患者、大多数家族性 ALS (FALS) 患者，以及所有伴有泛素阳性、tau 阴性包涵体（伴或不伴 ALS）的散发性和家族性 FTLD 患者中。最近在 FALS 患者中发现了 TDP-43 突变，表明突变 (MT) TDP-43 明显直接参与 ALS。 TDP-43 的毒性机制尚不清楚；然而，细胞质聚集体的存在表明该蛋白的错误折叠在 ALS 的发病机制中很重要。这些聚集体可能会隔离对运动神经元 (MN) 活力很重要的 TDP-43 结合蛋白。在此应用中，我们计划使用噬菌体展示库来：（1）识别和表征结合 TDP-43 的肽并预测细胞相互作用蛋白； (2) 鉴定和表征可变区抗体 (scFv) 的单链片段，可用于阐明 TDP-43 的功能并可能具有治疗潜力。 公众健康相关性：肌萎缩侧索硬化症 (ALS) 是一种无法治愈且无法有效治疗的绝望疾病。该应用涉及识别肽配体并生成与蛋白质结合的抗体，该蛋白质似乎是 ALS 以及其他神经退行性过程发病机制的关键。肽配体可用于预测细胞相互作用蛋白质，而抗体可阐明蛋白质的功能。总之，这项工作可能会深入了解为什么这种蛋白质的突变会导致 ALS，并可能为散发性和遗传性 ALS（以及其他神经退行性疾病）的治疗提供方向。