Molecular phenotyping of cortical cell types in multiple rodent models of ALS

多种 ALS 啮齿动物模型中皮质细胞类型的分子表型

基本信息

批准号：
9258507
负责人：
Eric F Schmidt
金额：
$ 37.08万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-01 至 2021-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9258507
关键词：
Address Affinity Chromatography American Amyotrophic Lateral Sclerosis Anatomy Animal Model Architecture Astrocytes Basic Science Bioinformatics Brain Brain Stem Candidate Disease Gene Cell Nucleus Cells Cerebral cortex Clinical Collaborations Diagnosis Disease Disease Progression Disease model Early Intervention Engineering Familial Amyotrophic Lateral Sclerosis Gene Expression Genes Genetic Techniques Genetic study Genome Histologic Human Knowledge Label Lead Link Methodology Methods Modeling Molecular Molecular Analysis Molecular Genetics Motor Motor Neuron Disease Motor Neurons Mouse Strains Mus Muscle Mutation Nerve Degeneration Neurons New York Paralysed Pathologic Processes Pathology Pathway interactions Patients Play Population Process Proteins Pyramidal Cells RNA purification Report (document)Research Research Project Grants Resources Ribosomes Rodent Model Role Signal Pathway Spinal Cord Therapeutic Time Transgenes Transgenic Mice Transgenic Organisms Translating Translations Universities Vulnerable Populations astrogliosis cell type comparative experimental study improved innovation molecular pathology molecular phenotype motor neuron degeneration mouse model mutant neuron component neuropathology new therapeutic target novel pre-clinical pre-clinical research programs protein TDP-43 public health relevance response success superoxide dismutase 1 therapeutic candidate tool transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): Over 30,000 Americans currently suffer from amyotrophic lateral sclerosis (ALS) which is characterized by progressive paralysis due to the degeneration of nerve cells in the brain and spinal cord that control muscles. Almost all cases of ALS are eventually fatal and the rapid progression of the disease makes it particularly terrible, with over 80% of patients dying within five years of diagnosis. No cure exists for ALS and the only available treatment slows disease progression by merely a few months. Therefore a great need exists for more effective and specific therapies that can stop or even reverse neurodegeneration. Innovation for such therapies will only arise from a better understanding of the molecular mechanisms underlying the pathological process. The proposed study aims to identify molecules and pathways dysregulated during disease progression in specific cell populations in the cerebral cortex, including the vulnerable "upper" motor neurons (UMNs). Such an analysis has never been done before due to the complexity of cortical architecture hampering the ability to distinguish between cell populations. Genetic studies have linked a number of genes to ALS pathology, including SOD1, TDP43, and FUS, yet all of these genes are widely expressed in many cell types throughout the body while ALS afflicts only certain cells in the CNS. This project will utilize the novel translating ribosome affinity purification (TRAP) methodology to overcome these limitations by allowing for the examination of protein translation from genetically defined cell types. Engineered mice harboring the TRAP transgene (bacTRAP mice) in four cortical cell types (two populations of vulnerable UMNs, a non-vulnerable neuronal population, and astrocytes) will be crossed to three mouse models of ALS that utilize disease-linked mutations in the SOD1 (G93A), TDP43 (M337V), and FUS (P525L) genes. These models recapitulate the neurodegeneration seen in human patients and will enable a comprehensive assessment of cell-type specific molecular changes during ALS pathology. Changes in gene expression during disease progression will be determined by analyzing TRAP translational profiles at three time points (early, pre-symptomatic, and late) within each model. While this is a pre-clinical basic research project, efforts will be focused on identifying candidate genes that wil have the strongest and most immediate clinical impact. Particular emphasis will be placed on changes that occur at early and pre-symptomatic stages since earlier intervention will likely have an increased rate of success. These studies aim to improve upon the success rate of therapies arising from animal models by probing genes altered specifically in vulnerable cells across multiple models. Results from the proposed study will provide the field with a valuable resource of novel genes and signaling pathways to serve as candidate targets for more specific and innovative therapeutics to treat ALS.

描述（由申请人提供）：目前有超过 30,000 名美国人患有肌萎缩侧索硬化症 (ALS)，其特征是由于大脑和脊髓中控制肌肉的神经细胞退化而导致进行性瘫痪，几乎所有 ALS 病例最终都会致命。而且这种疾病的快速进展使其变得尤为可怕，超过 80% 的患者在诊断后五年内死亡。 ALS 尚无治愈方法，唯一可用的治疗方法只能减缓疾病进展几个小时。因此，迫切需要能够阻止甚至逆转神经变性的更有效和特异性的疗法，只有更好地了解病理过程的分子机制，才能产生这种疗法的创新。由于大脑皮层结构的复杂性阻碍了区分细胞群的能力，因此以前从未进行过此类分析。研究发现许多基因与ALS 病理学，包括 SOD1、TDP43 和 FUS，但所有这些基因在全身多种细胞类型中广泛表达，而 ALS 仅影响 CNS 中的某些细胞。该项目将利用新型翻译核糖体亲和纯化 (TRAP) 方法。通过允许检查基因定义的细胞类型的蛋白质翻译来克服这些限制，这些小鼠在四种皮质细胞类型（两个易受影响的群体）中携带TRAP转基因小鼠（bacTRAP小鼠）。 UMN（非脆弱神经元群体和星形胶质细胞）将与三种 ALS 小鼠模型杂交，这些模型利用 SOD1 (G93A)、TDP43 (M337V) 和 FUS (P525L) 基因中的疾病相关突变。这些模型概括了这些模型。人类患者中观察到的神经退行性变将能够通过分析 TRAP 来确定 ALS 病理过程中细胞类型特异性分子变化。每个模型中三个时间点（早期、症状前和晚期）的翻译概况。在临床前基础研究项目中，工作重点将集中于确定具有最强和最直接临床影响的候选基因，并将特别重视早期和症状前阶段发生的变化，因为早期干预可能会增加。这些研究旨在通过探测多个模型中易受影响的细胞中特异改变的基因来提高动物模型治疗的成功率，该研究的结果将为该领域提供新基因和信号通路的宝贵资源。作为更具体和更具体的候选目标治疗 ALS 的创新疗法。