Mechanisms of Neurodegeneration in KIF5A ALS/FTD

KIF5A ALS/FTD 神经退行性变的机制

基本信息

批准号：
10740732
负责人：
Jonathan Robert Brent
金额：
$ 20.82万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2028-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10740732
关键词：
ALS patients Adaptor Signaling Protein Affect Age Amyotrophic Lateral Sclerosis Applications Grants Axon Axonal Transport Behavior Behavioral Binding Biochemical Biology Brain C-terminal CRISPR/Cas technology Cells Cessation of life Complex Cytoplasmic Granules Cytoskeleton Data Defect Development Development Plans Disease Distal Doctor of Medicine Doctor of Philosophy Exclusion Exons Fluorescence Microscopy Foundations Frontotemporal Dementia Functional disorder Funding Future Genetic Genetic Engineering Genetically Engineered Mouse Gliosis Goals Health Homeostasis Human Hyperactivity Image Imaging Techniques In Vitro Induced pluripotent stem cell derived neurons Kinesin Knowledge Label Language Longevity Lysosomes Maintenance Manuscripts Mentors Mentorship Microtubules Mitochondria Morphology Motor Motor Neuron Disease Motor Neurons Mus Mutation Nerve Degeneration Neurites Neurodegenerative Disorders Neuromuscular Junction Neurons Organelles Pathogenesis Pathogenicity Pathology Patients Pattern Physicians Post-Translational Protein Processing Preparation Production Proteins Proteomics Publications RNA RNA Interference RNA Splicing Records Regulation Research Resources Role Scientist Site Slide Solid Spinal Cord Splice-Site Mutation Structure Synapses Techniques Testing Therapeutic Time Tissues Training Transmission Electron Microscopy Universities Western Blotting Work age related aged career development causal variant cell motility cognitive function cohort confocal imaging frontotemporal lobar dementia amyotrophic lateral sclerosis gain of function gain of function mutation genetic regulatory protein human disease in vivo induced pluripotent stem cell insight live cell imaging mouse model mutant neurofilament neuron loss novel programs protein TDP-43 protein aggregation rational design skills small molecule tool trafficking ultra high resolution

项目摘要

Amyotrophic Lateral Sclerosis (ALS) is a fatal disease characterized by the dysfunction and death of motor neurons. Some ALS patients develop widespread neuronal damage leading to Frontotemporal Dementia (FTD), which is characterized by progressive behavioral change and language decline. Significant overlap exists between ALS and FTD regarding the genetic causes, suggesting that they may share common pathogenic mechanisms. Recently, mutations in motor protein, KIF5A, were identified in patients with ALS, ALS/FTD, and FTD alone. These concentrate in the splicing regulatory sites flanking exon 27 leading to its exclusion and abnormal KIF5A C-terminal domain structure. Our preliminary findings suggest that KIF5A ALS/FTD mutations result in the production of a constitutively active protein that causes disease through toxic gain of function. This proposal will test the hypothesis that KIF5A ALS/FTD is caused by gain of function of KIF5A activity in axonal transport and cytoskeletal regulation. To ensure that our work will be highly disease relevant we will utilize human iPSC derived motor neurons and mouse models harboring KIF5A ALS/FTD mutation. In Aim 1: we will define the effects of KIF5A ALS mutations on the assembly and motility of motor/cargo complexes. Specifically, we will utilize live-cell imaging of cargo motility, assessment of cargo distribution by IHC and transmission electron microscopy (TEM), candidate-based biochemical assessment of interactions with cargo adaptors, and unbiased assessment of interactions with potentially novel cargo adaptors and regulatory proteins using proximity labelling. In Aim 2: we will characterize the effects of KIF5A ALS/FTD mutation on cytoskeletal structure using IHC and TEM, live-imaging of MT dynamics and sliding, and analysis of MT post-translational modifications. In Aim 3: we will develop a mouse model of KIF5A ALS/FTD and perform analysis of longevity, behavior, and pathology. The mouse model will be an invaluable resource to validate findings from our in vitro studies and to advance knowledge regarding key features of neurodegeneration in KIF5A ALS/FTD. My ultimate goal is to become a successful physician-scientist with an independent research program investigating the mechanisms of neurodegeneration in motor neuron disease with a specific focus on axonal transport and cytoskeletal dynamics. Northwestern University has unique strengths in the study of ALS, FTD, and cytoskeletal biology and an outstanding commitment to the development of physician-scientists. The mentorship team includes renowned scientists with strong records in mentorship including Han-Xiang Deng M.D., Ph.D. (Primary Mentor) and Robert Kalb M.D (co-mentor). The career development plan focuses on broadening the awardee’s portfolio of research publications and presentations, which will be necessary for successful competition for RO1 funding. Career development activities will bolster grantsmanship and manuscript preparation skills and provide extensive didactic training in advanced imaging techniques, proteomics, cytoskeletal biology and ALS/FTD degenerative mechanisms.

肌萎缩侧索硬化症（ALS）是一种致命性疾病，其特征是运动功能障碍和死亡一些 ALS 患者出现广泛的神经元损伤，导致额颞叶痴呆 (FTD)，其特点是进行性行为改变和语言衰退存在显着的重叠。 ALS 和 FTD 之间关于遗传原因的研究表明，它们可能具有共同的致病因素最近，在 ALS、ALS/FTD 和 ALS 患者中发现了运动蛋白 KIF5A 的突变。仅 FTD。这些集中在外显子 27 侧翼的剪接调控位点，导致其被排除和我们的初步研究结果表明 KIF5A ALS/FTD 突变。导致产生一种组成型活性蛋白，该蛋白通过毒性功能获得而引起疾病。该提案将检验 KIF5A ALS/FTD 是由轴突中 KIF5A 活性功能增强引起的假设为了确保我们的工作与疾病高度相关，我们将利用人类。 iPSC 衍生的运动神经元和携带 KIF5A ALS/FTD 突变的小鼠模型在目标 1 中：我们将定义。具体来说，我们将研究 KIF5A ALS 突变对运动/货物复合体的组装和运动的影响。利用货物运动的活细胞成像，通过 IHC 和传输电子评估货物分布显微镜（TEM），基于候选的与货物适配器相互作用的生化评估，以及公正的使用邻近标记评估与潜在新颖的货物接头和调节蛋白的相互作用。在目标 2 中：我们将使用 IHC 和 IHC 来表征 KIF5A ALS/FTD 突变对细胞骨架结构的影响 TEM、MT 动力学和滑动的实时成像以及 MT 翻译后修饰的分析在目标 3 中：我们。将开发 KIF5A ALS/FTD 小鼠模型并进行寿命、行为和病理学分析。小鼠模型将是验证我们体外研究结果并推进研究的宝贵资源关于 KIF5A ALS/FTD 神经退行性疾病主要特征的知识我的最终目标是成为一名成功的医师科学家，拥有独立的研究计划，调查机制运动神经元疾病中的神经变性，特别关注轴突运输和细胞骨架动力学。西北大学在 ALS、FTD 和细胞骨架生物学研究方面具有独特的优势，导师团队包括著名的医师科学家的发展。具有良好指导记录的科学家包括邓汉翔医学博士、博士（主要导师）和罗伯特 Kalb M.D（联合导师）。职业发展计划的重点是扩大获奖者的研究范围。出版物和演示文稿，这对于成功竞争 RO1 职业资助是必要的。开发活动将加强资助和手稿准备技能，并提供广泛的先进成像技术、蛋白质组学、细胞骨架生物学和 ALS/FTD 退行性病变的教学培训机制。