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）：该应用的重点是TDP-43的异常，该蛋白是多种神经退行性疾病的基础（额叶lobar痴呆[FTLD]，帕金森氏病[PD]和疗效的后期孢子虫[ALS]）。 2006年，发现FTLD中神经元中的泛质细胞夹杂物包含聚集的TDP-43的异常磷酸化片段，TDP-43是一种通常在位置的DNA结合蛋白。值得注意的是，在所有零星ALS，大多数家族ALS（FALS）的患者以及所有患有偶然和家族性FTLD的患者患有泛素阳性，tau阴性的内含物，有或没有ALS的所有患者中，都可以看到TDP-43夹杂物。最近在FALS患者中鉴定出TDP-43突变，表明突变体（MT）TDP-43在ALS中明显直接受累。 TDP-43毒性的基础机制尚不清楚；然而，细胞质聚集体的存在表明该蛋白的错误折叠在ALS的发病机理中很重要。这些聚集体可能会隔离TDP-43结合蛋白对运动神经元（MN）的生存能力很重要。在此应用程序中，我们计划使用噬菌体显示库来：（1）识别和表征结合TDP-43并预测细胞相互作用蛋白的肽； （2）识别和表征可变区域抗体（SCFV）的单链片段，可用于阐明TDP-43的功能并具有治疗潜力。 公共卫生相关性：肌萎缩性横向硬化症（ALS）是一种绝望的疾病，没有治愈方法，也没有有效的治疗。该应用涉及鉴定肽配体并生成结合蛋白质的抗体，该蛋白似乎是ALS发病机理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的蛋白质，抗体可以阐明蛋白质的功能。总之，这项工作可以洞悉为什么该蛋白质引起ALS的原因，并有可能为零星和遗传的ALS（以及其他神经退行性疾病）的治疗提供了方向。