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是一种导致许多神经退行性疾病（额颞叶痴呆[FTLD]、帕金森病[PD]和肌萎缩侧索硬化[ALS]）的蛋白质。在2006年，发现FTLD神经元中的泛素化细胞质内含物含有聚集的TDP-43的异常磷酸化片段，TDP-43是一种通常位于核内的DNA结合蛋白。值得注意的是，TDP-43包涵体见于所有散发性ALS患者、大多数家族性ALS患者（FALS）以及所有散发性和家族性FTLD患者，这些患者具有泛素阳性、tau阴性包涵体，伴或不伴ALS。TDP-43突变最近已在ALS患者中鉴定，表明突变型（MT）TDP-43在ALS中的明确直接参与。TDP-43毒性的潜在机制尚不清楚;然而，细胞质聚集体的存在表明该蛋白质的错误折叠在ALS的发病机制中很重要。这些聚集体可以螯合对运动神经元（MN）的活力重要的TDP-43结合蛋白。在本申请中，我们计划使用噬菌体展示文库：（1）鉴定和表征结合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，其可用于阐明TDP-43的功能并可能具有治疗潜力。 公共卫生相关性：肌萎缩侧索硬化症（ALS）是一种绝望的疾病，目前没有治愈和有效的治疗。该应用涉及识别肽配体并产生与蛋白质结合的抗体，该蛋白质似乎是ALS发病机制以及其他神经退行性过程的关键。肽配体可用于预测细胞相互作用蛋白质，抗体可阐明蛋白质的功能。总之，这项工作可以深入了解这种蛋白质突变导致ALS的原因，并可能为治疗散发性和遗传性ALS（以及其他神经退行性疾病）提供方向。