Role of BET proteins in neuroinflammation in tauopathy

BET 蛋白在 tau 蛋白病神经炎症中的作用

• 批准号: 10512035
• 负责人: Edward Alan Burton
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-10-01 至 2025-09-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10512035
• 关键词: Acetylation; Address; Affinity; Alzheimer' s Disease; Animal Model; Autopsy; Binding; Biochemical; Biological Models; Brain; Brain Injuries; Bromodomain; CRISPR/Cas technology; Cells; Chemicals; Chronic; Clinical Trials; Complement; Craniocerebral Trauma; Cytoplasm; Data; Deterioration; Development; Disease; Disease Progression; Encephalitis; Event; Exposure to; Flow Cytometry; Frontotemporal Dementia; Funding; Gene Expression Profiling; Genetic; Genetic Transcription; Gliosis; Glutathione; Health; Human; Infiltration; Inflammatory; Length; Link; Lysine; Measures; Mediating; Microglia; Modeling; Molecular; Morphology; Motor; Natural Immunity; Nerve Degeneration; Nervous System Physiology; Neurodegenerative Disorders; Neurologic; Neurologic Deficit; Neurologic Dysfunctions; Neurons; Oxidative Stress; Pathogenesis; Pathogenicity; Pathologic; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Phosphorylation; Play; Process; Progressive Supranuclear Palsy; Proteins; Recovery; Reporter; Role; Shapes; Signal Transduction; Small Molecule Chemical Library; Symptoms; Synapses; Tauopathies; Testing; Therapeutic; Toxic effect; Traction; Transgenic Mice; Transgenic Organisms; Veterans; Work; Zebrafish; brain cell; brain tissue; cancer clinical trial; chronic traumatic encephalopathy; clinical development; comparative efficacy; density; disability; effective therapy; glial activation; hyperphosphorylated tau; in vivo; inhibitor; innovation; intravital imaging; microscopic imaging; migration; mutant; neuroinflammation; neuron loss; new therapeutic target; novel; novel strategies; oculomotor; oxidation; prevent; programs; ratiometric; response; selective expression; synaptic pruning; tau Proteins; tau aggregation; tau expression; therapeutic target; translational potential

Tauopathies are a group of neurodegenerative diseases that are characterized pathologically by neuronal loss and the accumulation of aggregates of the microtubule-associated protein Tau in surviving CNS neurons. Collectively, these disease, which include progressive supranuclear palsy (PSP), frontotemporal dementia (FTD), chronic traumatic encephalopathy (CTE) and Alzheimer's disease (AD), are a common and important cause of progressive neurological disability in veterans. Current treatments only partially mitigate symptoms and do not alter the course of these diseases; consequently, there is an urgent need for new approaches that address pathogenic mechanisms and prevent disease progression. Convergent genetic and pathological evidence strongly suggests that Tau is centrally involved in causing neurodegeneration in these diseases, and there is accumulating evidence that microglial activation and pathological microglial synaptic pruning in response to Tau accumulation plays a key role in pathogenesis. However, there is limited understanding of the mechanisms regulating aberrant functions of microglia in tauopathy and whether this could be targeted by novel therapies. During the current funding period, we discovered that a chemical inhibitor of bromodomains (protein modules that bind acetylated lysine residues) rescues both neurological function and neuroinflammation in a novel zebrafish tauopathy model. By using CRISPR/Cas9 to make zebrafish genetic null mutants, we found that the chemical targets a bromodomain and extraterminal motif-containing (BET) protein, Brd4, which has previously been implicated in pro-inflammatory signaling. Building on this work, our overall guiding hypothesis is that: Brd4 is necessary for microglial activation and synapse elimination in tauopathy and is a valid therapeutic target for treatment of these diseases. To address this hypothesis directly in vivo, we have crossed our novel zebrafish tauopathy model with a transgenic line expressing a fluorescent reporter protein in its microglia, allowing direct intravital microscopic imaging of microglial dynamics in the intact brain in vivo (Aim 1A) and recovery of microglia by flow cytometry for gene expression analysis (Aim 1B). We will determine how microglial functions and transcriptional programs are altered in tauopathy and by modulation of Brd4. We will further restore Brd4 expression within microglia of Brd4-null zebrafish to establish whether critical functions of Brd4 in tauopathy are cell-autonomous to microglia (Aim 1C). We will next determine how Brd4 influences neuronal tauopathy, oxidative stress and synaptic integrity using intravital imaging and biochemical approaches (Aim 2). Finally, we will test all available bromodomain inhibitors, including some that have already been used in human clinical trials, for activity in preventing neurological and neuroinflammatory phenotypes in the zebrafish tauopathy model (Aim 3A). We will take the most promising candidate into a mammalian tauopathy model to test its translational potential (Aim 3B). Together, these studies will elucidate the role of Brd4 in tauopathy and provide unique and innovative translational opportunities for development of Brd4-directed therapeutics.

Tauopathies是一组以神经元丢失为病理特征的神经退行性疾病 以及微管相关蛋白Tau聚集体在存活的CNS神经元中的积累。 总的来说，这些疾病，包括进行性核上性麻痹（PSP），额颞叶痴呆（FTD）， 慢性创伤性脑病（CTE）和阿尔茨海默病（AD）是阿尔茨海默病的常见和重要原因。 进行性神经功能障碍目前的治疗只能部分缓解症状， 改变这些疾病的进程;因此，迫切需要新的方法来解决 致病机制和预防疾病进展。遗传和病理学证据一致 表明Tau在这些疾病中主要参与引起神经变性， 有证据表明，小胶质细胞活化和病理性小胶质细胞突触修剪响应于Tau积累 在发病机制中起关键作用。然而，对异常的细胞凋亡的调控机制的了解有限。 小胶质细胞在tau蛋白病中的功能，以及这是否可以通过新的疗法来靶向。于本 在资助期间，我们发现了一种溴结构域（结合乙酰化的蛋白质模块）的化学抑制剂 赖氨酸残基）在新的斑马鱼tau蛋白病模型中拯救神经功能和神经炎症。 通过使用CRISPR/Cas9来制造斑马鱼遗传无效突变体，我们发现这种化学物质靶向一种 溴结构域和含有末端外基序（BET）的蛋白，Brd 4，它以前曾被牵连 在促炎信号中的作用。在这项工作的基础上，我们的总体指导假设是：Brd 4是必要的 对于tau蛋白病中的小胶质细胞活化和突触消除，并且是用于治疗的有效治疗靶点 这些疾病。为了直接在体内解决这一假设，我们将我们的新斑马鱼tau蛋白病 在其小胶质细胞中表达荧光报告蛋白的转基因系的模型，允许直接活体内 体内完整脑中小胶质细胞动力学的显微镜成像（Aim 1A）和 流式细胞术用于基因表达分析（Aim 1B）。我们将确定小胶质细胞的功能， 在Tau蛋白病中和通过调节Brd 4改变转录程序。我们将进一步恢复Brd 4 在Brd 4缺失的斑马鱼的小胶质细胞内表达，以确定Brd 4在tau蛋白病中是否具有关键功能 对小胶质细胞是细胞自主的（Aim 1C）。我们接下来将确定Brd 4如何影响神经元tau蛋白病， 氧化应激和突触完整性使用活体成像和生物化学方法（目的2）。最后我们 将测试所有可用的溴结构域抑制剂，包括一些已经用于人类临床的药物。 试验，用于预防斑马鱼tau蛋白病的神经和神经炎症表型的活性 模型（Aim 3A）。我们将把最有希望的候选者带到哺乳动物tau蛋白病模型中，以测试其 翻译电位（Aim 3B）。总之，这些研究将阐明Brd 4在tau蛋白病中的作用，并提供 为开发Brd 4导向疗法提供了独特和创新的转化机会。