Phage Display Investigations of TDP-43

描述(申请人提供)：本申请的重点是TDP-43的异常，这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]和肌萎缩侧索硬化症[ALS])的蛋白质。2006年，在FTLD的神经元中发现泛素胞质包涵体含有聚集的TDP-43的异常磷酸化片段，TDP-43是一种DNA结合蛋白，位于正常核的位置。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，以及所有伴有或不伴有ALS泛素阳性、tau阴性包涵体的散发性和家族性FTLD患者。最近在FAL患者中发现了TDP-43突变，表明突变型(MT)TDP-43明显直接参与了ALS。TDP-43的毒性机制尚不清楚；然而，胞浆聚集体的存在表明该蛋白的错误折叠在ALS的发病机制中起重要作用。这些聚集体可以隔离TDP-43结合蛋白，这些蛋白对运动神经元(MN)的生存至关重要。在这一应用中，我们计划使用噬菌体展示文库来：(1)鉴定和鉴定与TDP-43结合的多肽并预测细胞相互作用蛋白；(2)鉴定和鉴定可用于阐明TDP-43功能并可能具有治疗潜力的可变区抗体(ScFv)的单链片段。 公共卫生相关性：肌萎缩侧索硬化症(ALS)是一种无法治愈也没有有效治疗方法的绝症。这一应用涉及识别多肽配体和产生抗体，以结合一种蛋白质，该蛋白质似乎是ALS和其他神经退化过程的关键。多肽配体可用于预测与细胞相互作用的蛋白质，抗体可用于阐明蛋白质的功能。总之，这项工作可能为了解这种蛋白突变导致ALS的原因提供洞察力，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。

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).