Developing RNA therapeutics for TDP-43 proteinopathy in ALS/FTD

开发针对 ALS/FTD 中 TDP-43 蛋白病的 RNA 疗法

基本信息

批准号：
10534538
负责人：
Katie Elizabeth Copley
金额：
$ 4.68万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10534538
关键词：
ALS patients Affect Alzheimer&apos s disease related dementia Amyotrophic Lateral Sclerosis Behavior Binding Biochemical Biological Assay C-terminal Cell model Cells Cessation of life Clinical Data Data Analyses Development Disease Electron Microscopy Environment Etiology Evolution Exhibits Fibroblasts Fostering Frontotemporal Dementia Goals Health In Vitro Laboratories Language Link Missense Mutation Modeling Molecular Motor Neurons Nerve Degeneration Neurodegenerative Disorders Neurons Neurosciences Nucleotides Pathologic Pathology Patients Pennsylvania Personality Physiology RNA RNA Binding RNA Splicing RNA-Binding Proteins Recombinants Research Personnel Role Solubility Specificity Symptoms TDP-43 aggregation Techniques Temporal Lobe Testing Therapeutic Toxic effect Training Universities Variant base career cellular pathology collaborative environment crosslinking and immunoprecipitation sequencing design disease phenotype disorder subtype experimental study frontal lobe frontotemporal lobar dementia-amyotrophic lateral sclerosis gain of function in vitro Assay insight motor impairment mutant neuron loss prevent protein TDP-43 stem success therapeutic RNA therapeutically effective transcriptome sequencing

项目摘要

PROJECT SUMMARY My long-term career goal is to tackle neurodegeneration as an academic researcher. Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), an Alzheimer's Disease Related Dementia (ADRD), are both fatal neurodegenerative disorders characterized by neuronal loss. ALS is primarily characterized by motor impairments stemming from loss of motor neurons, whereas the main symptoms of FTD include changes in personality, behavior, and language stemming from loss of cortical neurons in the frontal and temporal lobes. ALS and FTD exist on a disease spectrum, where some patients present with features of both diseases. A molecular hallmark shared by almost all ALS patients and approximately half of FTD patients is the pathological aggregation of the RNA-binding protein TDP-43. It has recently been established that the solubility of TDP-43 is increased by its binding to RNA. My preliminary data utilizing in vitro aggregation assays with purified TDP-43 indicate that missense mutations in TDP-43 can alter the ability of RNA to prevent aggregation of TDP-43. Based on these findings, I hypothesize that aberrant aggregation of TDP-43 in ALS/FTD models is due to alterations in TDP-43:RNA interactions. and that directly manipulating the RNA interactions of TDP-43 can rescue disease phenotypes. The goal of this proposal is to assess if differences in the RNA interactions of TDP-43 are present in disease by 1) examining the ability of short RNAs to prevent aggregation of disease-linked TDP-43 variants in vitro and 2) determining if there are alterations in the RNAs that TDP-43 binds to in ALS/FTD patient-derived neurons versus control neurons. The proposal also aims to 3) determine if administration of short RNAs that bind TDP-43 can rescue disease phenotypes in ALS/FTD patient-derived neurons. The proposed experiments will provide critically lacking information on the interactions between TDP-43 and RNA in disease as well as strategies to target these interactions therapeutically. The proposed experiments will substantially contribute to my training, allowing me to gain expertise in new techniques such as electron microscopy, crosslinking immunoprecipitation (CLIP), and sequencing data analysis. My training environment will foster success for the proposal, combining the biochemical expertise of the Shorter laboratory and the exceedingly collaborative environment at the University of Pennsylvania and within the Neuroscience Graduate Group. These studies will facilitate both my scientific and career goals by supporting my evolution into an academic researcher who develops therapeutics for patients with neurodegenerative disease.

项目摘要我的长期职业目标是应对作为学术研究人员的神经退行性。肌萎缩性外侧硬化症（ALS）和额颞痴呆（FTD），阿尔茨海默氏病有关的痴呆症（ADRD）都是由神经元丧失的致命神经退行性疾病。 ALS的主要特征是运动神经元的丧失导致运动障碍，而FTD的主要症状包括额叶和颞叶中皮质神经元丧失的人格，行为和语言变化。 ALS和FTD存在于疾病范围内，其中一些患者均具有两种疾病的特征。几乎所有ALS患者和大约一半的FTD患者共享的分子标志是RNA结合蛋白TDP-43的病理聚集。最近已经确定，TDP-43的溶解度通过与RNA的结合增加了。我使用纯化的TDP-43的体外聚集测定法的初步数据表明，TDP-43中的错义突变可以改变RNA防止TDP-43聚集的能力。基于这些发现，我假设ALS/FTD模型中TDP-43的异常聚集是由于TDP-43：RNA相互作用的改变。直接操纵TDP-43的RNA相互作用可以挽救疾病表型。该提案的目的是评估疾病中TDP-43的RNA相互作用的差异是否存在1）检查短RNA防止疾病连接的TDP-43变体在体外和2）确定TDP-43与TDP-43在ALS/FTD NEURONS中结合的RNA是否发生变化的能力。该提案还旨在3）确定结合TDP-43的短RNA是否可以挽救ALS/FTD患者衍生的神经元中的疾病表型。拟议的实验将为疾病中TDP-43与RNA之间的相互作用以及以治疗靶向这些相互作用的策略提供严重的信息。提出的实验将有助于我的培训，从而使我能够获得新技术的专业知识，例如电子显微镜，交联免疫沉淀（剪辑）和测序数据分析。我的培训环境将促进该提案的成功，结合宾夕法尼亚大学和神经科学研究生群体的较短实验室和极其协作环境的生化专业知识。这些研究将通过支持我发展为神经退行性疾病患者的学术研究人员的进化来促进我的科学和职业目标。