Genes Mediating Innate Immune Suppression by Hypercapnia in Mammals and Flies

哺乳动物和果蝇高碳酸血症介导先天免疫抑制的基因

• 批准号: 8238710
• 负责人: GREG J BEITEL
• 金额: $ 37.02万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-12-15 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8238710
• 关键词: Acute; Adult; Adverse effects; Bacterial Infections; Binding; Blood; CCL2 gene; Candidate Disease Gene; Carbon Dioxide; Cause of Death; Cells; ChIP-seq; Chromatin; Chronic; Chronic Obstructive Airway Disease; Clinical; Communities; Complex; Data; Defect; Drosophila genus; Foundations; Future; Gene Components; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Histones; Homologous Gene; Host Defense; Human; Hypercapnia; Immune; Immune Response Genes; Immunosuppression; Immunosuppressive Agents; In Vitro; Individual; Infection; Inflammatory; Interleukin-6; Lung; Lung diseases; Mammals; Mediating; Mediator of activation protein; Molecular; Mus; Mutation; Natural Immunity; Orthologous Gene; Outcome; Pathway interactions; Patients; Phagocytes; Play; Pneumonia; Predisposition; Proteins; Pseudomonas aeruginosa; RNA Interference; Regulator Genes; Risk; Risk Factors; Role; System; TNFRSF5 gene; Testing; Time; Tissues; Validation; Zinc Fingers; antimicrobial peptide; bacterial resistance; base; fly; genome-wide; histone methyltransferase; homeodomain; in vivo; macrophage; mortality; mouse model; p65; prevent; promoter; response; transcription factor

DESCRIPTION (provided by applicant): Hypercapnia, elevation of PCO2 in blood and tissue, commonly occurs in severe acute and chronic lung disorders, such as chronic obstructive pulmonary disease (COPD). Patients with advanced COPD frequently develop bacterial lung infections, and hypercapnia is a risk factor for mortality in such individuals, as in those with community-acquired pneumonia. We have shown that hypercapnia suppresses transcription of innate immune response genes required for host defense in human, mouse and Drosophila cells, and increases mortality due to bacterial infections in mice and Drosophila. These findings suggest that hypercapnia is not simply a marker of advanced lung disease, but plays a causal role in poor clinical outcomes by increasing susceptibility to infection. Our data also strongly suggest that hypercapnia inhibits innate immunity and host defense by pathway(s) conserved from Drosophila to mammals. Because the molecular mediator(s) of hypercapnic immune suppression are undefined, we conducted a genome-wide RNAi screen in cultured Drosophila cells, which identified ~140 genes required for hypercapnic suppression of antimicrobial peptide (AMP) genes. The 5 most potent of these candidate CO2-mediators encode the zinc finger homeodomain transcription factor, Zfh2; a histone deactylase; a histone methyltransferase; a chromatin-associated Ig-repeat protein; and a Rac-interacting protein. None of these genes were previously known to have immunoregulatory function. Exciting new data indicate that mutations in zfh2 protect adult Drosophila against CO2-induced host defense defects. Thus, we hypothesize that Zfh2 and the proteins encoded by the other 4 candidate genes are components of pathway(s) by which hypercapnia suppresses innate immune gene expression and host defense in Drosophila, and that the mammalian orthologs of these genes mediate hypercapnic suppression of innate immune/host defense genes in mouse and human phagocytes. In the case of Zfh2 and its mammalian orthologs, ZFHX3 and ZFHX4, we further hypothesize that hypercapnia alters their abilities to bind target gene promoters or components of the NF-:B transcriptional complex, or modifies the transcriptional activity of these factors, thereby decreasing expression of specific innate immune/host defense genes. In the proposed studies, we will test these hypotheses in vitro using cultured fly, mouse and human macrophages, and in vivo using adult Drosophila and lung inflammatory cells obtained from mice with Pseudomonas aeruginosa pneumonia. These studies will for the first time define components of conserved pathway(s) by which hypercapnia impairs innate immunity and host defense, and determine their mechanisms of action. The results should lay the basis for future studies aimed at preventing hypercapnic immune suppression in patients with advanced lung disease. PUBLIC HEALTH RELEVANCE: Hypercapnia, or elevation of the level of carbon dioxide (CO2) in the body, commonly develops in people with advanced lung diseases. These individuals more frequently develop lung infections and have an increased risk of dying. This project will determine how hypercapnia leads to more and worse infections by determining the function of the first identified mediator of hypercapnic immune suppression, and by identifying additional components of immunosuppressive CO2-response pathways .

描述（由申请人提供）：高碳酸血症，血液和组织中PCO 2升高，通常发生于严重急性和慢性肺部疾病，如慢性阻塞性肺病（COPD）。晚期COPD患者经常发生细菌性肺部感染，高碳酸血症是这些患者死亡的危险因素，如社区获得性肺炎患者。我们已经表明，高碳酸血症抑制了人类，小鼠和果蝇细胞中宿主防御所需的先天免疫反应基因的转录，并增加了小鼠和果蝇细菌感染引起的死亡率。这些研究结果表明，高碳酸血症不仅是晚期肺部疾病的标志物，而且通过增加感染的易感性在不良临床结局中发挥因果作用。我们的数据还强烈表明，高碳酸血症通过从果蝇到哺乳动物的保守途径抑制先天免疫和宿主防御。由于高碳酸血症免疫抑制的分子介体尚不明确，我们在培养的果蝇细胞中进行了全基因组RNAi筛选，确定了高碳酸血症抑制抗菌肽（AMP）基因所需的~140个基因。这些候选CO2介质中最有效的5种编码锌指同源结构域转录因子Zfh 2;组蛋白脱乙酰酶;组蛋白甲基转移酶;染色质相关Ig重复蛋白;和Rac相互作用蛋白。此前已知这些基因均不具有免疫调节功能。令人兴奋的新数据表明，zfh 2突变保护成年果蝇免受CO2诱导的宿主防御缺陷。因此，我们假设Zfh 2和其他4个候选基因编码的蛋白质是高碳酸血症抑制果蝇先天免疫基因表达和宿主防御的途径的组分，并且这些基因的哺乳动物直系同源物介导小鼠和人吞噬细胞中先天免疫/宿主防御基因的高碳酸血症抑制。在Zfh 2及其哺乳动物直系同源物ZFHX 3和ZFHX 4的情况下，我们进一步假设高碳酸血症改变了它们结合靶基因启动子或NF-：B转录复合物组分的能力，或改变了这些因子的转录活性，从而降低了特异性先天免疫/宿主防御基因的表达。在拟议的研究中，我们将测试这些假设在体外培养的苍蝇，小鼠和人类巨噬细胞，并在体内使用成年果蝇和肺部炎症细胞从铜绿假单胞菌肺炎小鼠。这些研究将首次确定高碳酸血症损害先天免疫和宿主防御的保守途径的组成部分，并确定其作用机制。该结果为将来预防晚期肺病患者高碳酸血症免疫抑制的研究奠定了基础。 公共卫生关系：高碳酸血症，或体内二氧化碳（CO2）水平的升高，通常发生在晚期肺病患者中。这些人更经常发生肺部感染，死亡风险增加。该项目将确定高碳酸血症如何导致更多和更严重的感染，通过确定高碳酸血症免疫抑制的第一个确定的介质的功能，并通过确定免疫抑制CO2反应途径的其他组成部分。