PHF15, a potential repressor of inflammation in the brain and its relevance to Alzheimer's disease

PHF15，一种潜在的大脑炎症抑制剂及其与阿尔茨海默病的相关性

• 批准号: 10477295
• 负责人: Kaoru Saijo
• 金额: $ 19.43万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10477295
• 关键词: ATAC-seq; Age; Aging; Alzheimer' s Disease; Alzheimer' s disease risk; Amyloid beta-Protein Precursor; Animals; Binding; Binding Proteins; Biochemical; Biological Assay; Brain; Cell Culture System; Cell Line; ChIP-seq; Complement; Data; Disease; Drug Targeting; Environmental Risk Factor; Enzymes; Epigenetic Process; Failure; Family member; Gene Expression; Genes; Goals; High Fat Diet; Hippocampus (Brain); Histones; Human; Impaired cognition; In Vitro; Infection; Inflammasome; Inflammation; Inflammation Mediators; Inflammatory; Knock-out; Ligands; Lysine; Mass Spectrum Analysis; Mediating; Messenger RNA; Microglia; Modeling; Molecular; Multiprotein Complexes; Mus; Nerve Degeneration; Neuroglia; Output; Oxidative Stress; PHD Finger; Pathogenicity; Patients; Play; Prefrontal Cortex; Prevention; Proteins; Reporting; Repression; Role; Signal Transduction; Stimulus; Structure; Symptoms; System; Testing; Therapeutic; Toll-like receptors; Transcription Repressor; Up-Regulation; Viral Genes; Virus Diseases; age related; aged; base; chromatin remodeling; experimental study; genetic risk factor; healthy aging; histone modification; in vitro Model; in vivo; loss of function; mRNA Expression; member; mouse model; neuroinflammation; non-genetic; prevent; protein complex; stressor; therapy development; tissue culture; treatment strategy

Abstract While genetic risk factors for Alzheimer’s disease (AD) have been identified, around ninety percent of patients are sporadic cases. Therefore, in this proposal, we seek to understand the molecular mechanisms behind the establishment of sporadic cases of AD. Recently, we have found that a putative epigenetic regulator, PHD finger 15 (PHD15), is a transcriptional repressor of genes regulating inflammation in microglia. Indeed, knockout of Phf15 in mouse microglial cells was sufficient to increase pro-inflammatory mediators and anti-viral genes, of note, including amyloid precursor protein (APP) and complement factors, in the absence of any stimuli. Because the mRNA expression of PHF15 in human and mouse microglia is increased upon healthy aging, we hypothesize that PHF15 epigenetically represses age-induced inflammation in microglia and that failure to up-regulate PHF15 results in neuroinflammation and cognitive decline. To test our hypothesis, we will identify members of the protein complex containing PHF15 by mass-spectrometry using a human microglial cell line that we have established. After confirming protein binding using biochemical assays, we will perform functional assays by establishing gain- and loss-of-function cell lines to validate whether the identified complex proteins cooperate with PHF15 to repress inflammation. In addition to isolating the PHF15 protein complex, we will also try to identify non-genetic factors that prevent PHF15 mRNA up-regulation using both in vivo and in vitro models. Based on our preliminary data, we will focus on virus infection and high-fat diet for the in vivo experiments, as well as oxidative stress and infection mimics (ligands for toll-like receptors and inflammasomes) for the in vitro experiments. Overall, the goals of this proposal are to determine the molecular mechanism behind how PHF15 represses inflammation in microglia and to identify non-genetic factors that prevent up-regulation of PHF15 expression. These results may help us to better understand the causes of sporadic cases of AD and, thus, direct the development of treatment strategies to treat and/or prevent this disease.

抽象的 尽管已经确定了阿尔茨海默氏病（AD）的遗传危险因素，但约有90％ 患者是零星的病例。因此，在此提案中，我们试图了解分子机制 建立零星的AD案件的背后。最近，我们发现推定的表观遗传学 调节剂PhD手指15（PHD15）是小胶质细胞中基因调节性炎症的转录反射器。 实际上，小鼠小胶质细胞中PHF15的敲除足以增加促炎性介体和 抗病毒基因，包括淀粉样蛋白前体蛋白（APP）和补体因子，在没有的情况下 任何刺激。因为在人和小鼠小胶质细胞中PHF15的mRNA表达增加了 健康衰老，我们假设PHF15表观遗传反映了小胶质细胞和 未能上调PHF15导致神经炎症和认知能力下降。为了检验我们的假设， 我们将使用人的质谱法确定含有PHF15的蛋白质​​复合物的成员 我们已经建立的小胶质细胞系。在使用生化测定确认蛋白质结合后，我们将 通过建立功能丧失和功能丧失细胞系来验证是否已确定的功能测定 复杂的蛋白质与PHF15合作以反映注射。除了隔离PHF15蛋白 复杂，我们还将尝试确定使用两者在 体内和体外模型。根据我们的初步数据，我们将专注于病毒感染和高脂饮食 体内实验以及氧化应激和感染模拟物（Toll样受体的配体和配体 炎症）用于体外实验。总体而言，该提议的目标是确定分子 PHF15如何反映小胶质细胞注射的机制，并确定非遗传因素 防止PHF15表达的上调。这些结果可能有助于我们更好地了解 AD的零星病例，因此指导了治疗策略的制定以治疗和/或阻止这种情况 疾病。