Investigating mechanisms of pathogenesis in TBK1-associated amyotrophic lateral sclerosis

研究 TBK1 相关肌萎缩侧索硬化症的发病机制

基本信息

批准号：
9583748
负责人：
Cindy V Ly
金额：
$ 16.31万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9583748
关键词：
Adaptor Signaling Protein Affect Alzheimer&apos s Disease Alzheimer&apos s disease risk Amyotrophic Lateral Sclerosis Animal Disease Models Animal Model Astrocytes Astrocytosis Autophagocytosis Award Basic Science Behavioral Assay Biochemical C9ORF72 Cells Cessation of life Clinical Complex Disease Disease Pathway Embryo Environment Expression Profiling Familial Amyotrophic Lateral Sclerosis Fibroblasts Foundations Frontotemporal Dementia Functional disorder Genes Genetic Immune Impairment In Vitro Infection Inflammation Inflammation Mediators Inflammatory Inflammatory Response Pathway Innate Immune Response K-Series Research Career Programs Knockout Mice Lead Medicine Mentors Mentorship Messenger RNA Methods Microglia Motor Motor Neurons Mus Mutation Natural Immunity Nerve Degeneration Nervous system structure Neuraxis Neurodegenerative Disorders Neuroglia Neurologic Neuromuscular Diseases Neurons Nonsense-Mediated Decay Organelles PGRN gene Pathogenesis Pathogenicity Pathway interactions Patients Peripheral Phagocytes Phagocytosis Phenotype Phosphorylation Phosphotransferases Physicians Process Proteins Research Research Training Risk Role Scientist Signal Transduction TANK-binding kinase 1 TREM2 gene Tissues Training Training Support Translating Universities Variant Washington base cell type cytokine diagnostic biomarker effective therapy experimental study extracellular gene function genetic association knowledge base loss of function mRNA Expression motor neuron degeneration mouse model nervous system disorder neurogenetics neuromuscular neuron loss new therapeutic target novel pathogen programs protein expression proteostasis recruit response skills stem stressor therapeutic target tool

项目摘要

The purpose of this mentored career development award is to propel an early stage PI towards independence as a physician-scientist with a research program on mechanisms of amyotrophic lateral sclerosis (ALS) and clinical focus on neuromuscular medicine. ALS is a rapidly progressive neurologic disorder characterized by motor neuron degeneration that leads to death in 3-5 years and for which there are no effective treatments. TANK binding kinase 1 (TBK1) was recently uncovered as a novel causative gene in sporadic and familial ALS and frontotemporal dementia (FTD). However, the mechanisms by which variants in TBK1 because disease remain unclear. Under the guidance of Dr. Timothy Miller, I propose to investigate how disruptions in TBK1, a ubiquitously expressed kinase that functions at the intersection of two processes implicated in ALS pathogenesis, autophagy and innate immunity, lead to disease pathogenesis. Although a subset of TBK1 mutations cause truncation and reduced expression of the protein, the significance and pathogenicity of many missense variants remain uncharacterized. Notably, haploinsufficiency of microglial- derived genes such as progranulin (PGRN) and triggering receptor expressed on myeloid cells 2 (TREM2) has been shown to cause or increase risk for Alzheimer's disease, frontotemporal dementia, and ALS [10] by altering inflammatory signaling and phagocytic function. Given the contribution of non-cell-autonomous mechanisms in motor neuron cell death in ALS and known expression of TBK1 in neuronal and non-neuronal cell types, I will also examine the impact of TBK1 loss of function (LoF) in astrocytes and microglia, the primary immune cells of the central nervous system. In order to understand how TBK1 LoF influences manifestation of disease, an animal model of disease would be an incredibly valuable tool. Unfortunately, previously described TBK1 knock out mice are embryonic lethal which precluded an analysis of neurological phenotypes. Based on these considerations, I propose the a research program incorporating the following aims: 1) Determine the functional consequences of TBK1 missense and LoF variants on autophagy using biochemical and histopathological methods 2) Determine the effect of TBK1 LoF on phagocytosis, autophagy, and inflammatory signaling in astrocytes and microglia 3) Determine whether TBK1 causes ALS via cell autonomous or non-cell autonomous mechanisms by examining phenotypes of mice with selective loss of TBK1 in motor neurons, astrocytes, and microglia The experiments proposed above are expected to clarify the role of TBK1 in ALS pathogenesis and reveal novel therapeutic targets. The research training supported by the award will fortify my skill set and knowledge base for translating basic science discoveries to novel bedside treatment and diagnostic markers for ALS and potentially other neurodegenerative diseases.

这项指导的职业发展奖的目的是推动早期PI作为医生 - 科学家的独立性，并通过有关肌萎缩性侧面硬化症机制的研究计划（ALS）和神经肌肉医学的临床关注。 ALS是一种以运动神经元变性为特征的快速进行性神经系统疾病，在3 - 5年内导致死亡，并且没有有效的治疗方法。坦克结合激酶1（TBK1）最近被发现为零星和家族性ALS和额颞痴呆（FTD）中的一种新型致病基因。但是，由于疾病尚不清楚，因此TBK1中变异的机制。在蒂莫西·米勒（Timothy Miller）博士的指导下，我建议研究TBK1中的破坏是一种普遍表达的激酶，该激酶在与ALS发病机理，自噬和先天免疫的两个过程的交点上起作用，导致疾病发病机理。尽管TBK1突变的一部分会导致蛋白质的截断和表达降低，但许多错义变体的显着性和致病性仍然没有表征。值得注意的是，在髓样细胞2（TREM2）上表达的小胶质细胞衍生基因的单倍症（例如pRigranulin（PGRN）和触发受体）已被证明会导致或增加阿尔茨海默氏病，额颞痴呆症和ALS [10]的风险，而改变了炎症信号和炎症信号的风险。鉴于非细胞自治机制在ALS运动神经元细胞死亡中的贡献以及TBK1在神经元和非神经元细胞类型中的已知表达，我还将检查星形胶质细胞和小胶质细胞中TBK1功能丧失（LOF）的影响，中枢神经系统的主要免疫细胞。为了了解TBK1 LOF如何影响疾病的表现，疾病的动物模型将是一种非常有价值的工具。不幸的是，先前描述的TBK1敲除小鼠是胚胎致死的，这排除了神经系统型的分析。基于这些考虑，我提出了一项研究计划，该计划结合了以下目的：1）确定使用生化和组织病理学方法的TBK1错义和LOF变体对自噬的功能后果2）确定TBK1 LOF通过吞噬性，自噬和炎症信号的影响，确定TBK1 LOF对吞噬，自噬和炎症信号的作用1，并确定Sustrocytes 1 3）3）或通过检查运动神经元，星形胶质细胞和小胶质细胞中TBK1选择性丢失的小鼠的表型，或者通过非细胞自主机制进行检查，上述实验有望阐明TBK1在ALS发病机理中的作用并揭示新的治疗靶标。该奖项支持的研究培训将加强我的技能和知识库，以将基础科学发现转化为新颖的床边治疗和ALS和潜在的其他神经退行性疾病的诊断标记。