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

基本信息

批准号：
10289321
负责人：
Kaoru Saijo
金额：
$ 23.47万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
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/10289321
关键词：
ATAC-seq Age Aging Alzheimer&apos s Disease Alzheimer&apos s disease risk Amyloid beta-Protein Precursor Animals Antiviral Agents 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

项目摘要

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.

抽象的虽然阿尔茨海默病 (AD) 的遗传风险因素已被确定，但大约百分之九十的人因此，在本提案中，我们寻求了解其分子机制。 AD散发病例成立的背后最近，我们发现了一个推定的表观遗传。 PHD Finger 15 (PHD15) 调节因子是小胶质细胞炎症调节基因的转录抑制因子。事实上，敲除小鼠小胶质细胞中的 Phf15 足以增加促炎介质和值得注意的是，抗病毒基因，包括淀粉样前体蛋白（APP）和补体因子，在缺乏因为人类和小鼠小胶质细胞中 PHF15 的 mRNA 表达在任何刺激下都会增加。健康老龄化，我们认为 PHF15 通过表观遗传抑制小胶质细胞中年龄引起的炎症，未能上调 PHF15 会导致神经炎症和认知能力下降为了检验我们的假设，我们将使用人类质谱仪来鉴定含有 PHF15 的蛋白质复合物的成员在使用生化测定确认蛋白质结合后，我们将建立小胶质细胞系。通过建立功能获得和丧失功能的细胞系来进行功能测定，以验证所识别的除了分离 PHF15 蛋白外，复合蛋白还与 PHF15 协同抑制炎症。复杂的情况下，我们还将尝试使用这两种方法来识别阻止 PHF15 mRNA 上调的非遗传因素根据我们的初步数据，我们将重点关注病毒感染和高脂肪饮食。体内实验，以及氧化应激和感染模拟物（Toll 样受体的配体和总的来说，该提案的目标是确定分子。 PHF15 抑制小胶质细胞炎症背后的机制，并确定非遗传因素防止PHF15表达上调这些结果可能有助于我们更好地了解其原因。 AD 散发病例，因此指导制定治疗和/或预防这种情况的治疗策略疾病。