Epigenetic regulation of viral infection and replication by periodontal bacteria

牙周细菌病毒感染和复制的表观遗传调控

• 批准号: 8739642
• 负责人: Fengchun Ye
• 金额: $ 48.11万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-23 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8739642
• 关键词: AIDS/HIV problem; Acetylation; Actins; Acute; Bacteria; Biological Models; CD4 Positive T Lymphocytes; Cells; Complex; Cytomegalovirus; Cytoskeleton; Data; Dissociation; EZH2 gene; Endothelial Cells; Epidemiology; Epigenetic Process; Epithelial; Epithelial Cells; Excision; Exhibits; Fusobacterium nucleatum; Gene Expression; Genetic Transcription; Gram-Negative Anaerobic Bacteria; HIV; HIV Infections; Herpesviridae; Herpesviridae Infections; Histone Acetylation; Histones; Human; Human Herpesvirus 4; Human Herpesvirus 8; Individual; Infection; Kaposi Sarcoma; Lead; Libraries; Link; Lysine; Metabolic; Methylation; Microtubules; Oral; Oral cavity; Pathway interactions; Patients; Periodontal Diseases; Periodontitis; Pilot Projects; Play; Polycomb; Porphyromonas gingivalis; Prevalence; Promoter Regions; Proteins; Proteomics; Recurrence; Regulation; Role; Severities; Simplexvirus; T-Lymphocyte; Testing; Transcriptional Activation; Umbilical vein; Viral; Viral Genes; Virus; Virus Diseases; Volatile Fatty Acids; base; latent infection; longitudinal human study; lytic replication; novel; novel strategies; oral bacteria; oral infection; oral lesion; periodontopathogen; prevent; promoter; public health relevance; screening; small hairpin RNA; trafficking; tumor

DESCRIPTION (provided by applicant): The purpose of the present study is to investigate the regulation of the host cell epigenetic machinery by metabolic products from periodontal bacteria and the impact on enhancing viral replication and infection in the oral cavity of HIV patients. It s well established that HIV patients exhibit increased prevalence and severity of chronic periodontitis and endure more frequent infections by herpesviruses such as herpes simplex virus (HSV), Epstein Barr virus (EBV), and Kaposi sarcoma-associated herpesvirus (KSHV). The oral cavity is also a potential reservoir for latent HIV infection. Previous studies suggest tht periodontal bacteria promote lytic replication of both HIV and herpeviruses, resulting in recurrent HIV infection and oral lesions. Anaerobic gram- negative bacteria produce various short chain fatty acids (SCFAs) that inhibit class-1/2 histone deacetylases (HDACs) to promote histone hyperacetylation and viral gene expression. However, whether SCFAs impact other epigenetic regulators and whether they also enhance viral infection have not been studied and remain to be determined. Data from our pilot study indicate that SCFAs not only induce lytic replication of HIV and KSHV but also increase the infection rates of the two viruses in their respective target cells. Besides inhibition of class-1/2 HDACs, SCFAs also down regulate expression of the class-3 HDAC SIRT1 and the subunit EZH2 of the polycomb repressive complex (PRC2), leading to histone hyperacetylation and reduction of repressive histone methylation simultaneously. Based on these results, we hypothesize that SCFAs modulate multiple epigenetic regulators to promote lytic replication of both HIV and herpesviruses such as KSHV in the oral cavity. We believe that SCFAs also impact the host cytoskelton and transport pathways to make cells more permissive to viral infection. A high throughput shRNA library screening for epigenetic regulators involved in HIV transcriptional activation has led to our identification of a number of epigenetic regulators that appear to form multi-protein silencing complexes. To test our hypotheses, in AIM-I, we will perform proteomic analysis to identify the specific components of the epigenetic silencing complexes and examine the effects of SCFAs on expression of the regulators in oral epithelial, endothelial, and CD4+ T- cells. We will also confirm the roles of the SCFAs-regulated epigenetic regulators in transcriptional activation of both HIV and KSHV in our model systems. Previous studies suggest that lysine acetylation of cytoskeleton actins and microtubule plays important roles in viral entry and intracellular trafficking. In AIM-II, we will examine if and how SCFAs cause lysine acetylation of cytoskeleton actins and microtubule to increase viral infectivity, as a consequence of SCFAs suppression of both class-1/2 HDACs and class-3 HDAC SIRT1. To further test our hypotheses, in AIM-III, we will examine if oral epithelial and T cells from HIV-patients with severe periodontal disease indeed display SCFAs-related epigenetic changes detected in AIM-I, and whether the epigenetic changes correlate with viral replication and infection in the patients, by conducting cross sectional and longitudinal human studies. It is expected that the present study will reveal novel and common mechanisms that control replication and infection of both HIV and herpesviruses in the oral cavity of HIV patients. The results should provide a scientific basis for developing novel strategies to prevent and treat recurrent HIV and herpesviral infections in HIV/AIDS patients simultaneously.

描述（由申请人提供）：本研究的目的是调查牙周细菌的代谢产物对宿主细胞表观遗传机制的调节，并对HIV患者口腔口腔的病毒复制和感染的影响。众所周知，HIV患者表现出慢性牙周炎的患病率和严重程度的增加，并通过疱疹病毒（例如单纯疱疹）病毒（HSV），爱泼斯坦Barr病毒（EBV）和Kaposi Sarcoma相关的疱疹病毒（KSHV）忍受了更频繁的感染。口腔也是潜在的潜在艾滋病毒感染储层。先前的研究表明，牙周细菌促进了HIV和HERPEVIRAS的裂解复制，导致复发 艾滋病毒感染和口腔病变。厌氧革兰氏阴性细菌产生各种短链脂肪酸（SCFA），可抑制类1/2组蛋白脱乙酰基酶（HDAC）促进组蛋白高乙酰化和病毒基因表达。但是，SCFA是否会影响其他表观遗传调节剂以及是否还会增强病毒感染，并且尚未研究并保持待确定。来自我们的试点研究的数据表明，SCFA不仅诱导HIV和KSHV的裂解复制，而且增加了各自靶细胞中两种病毒的感染率。除了抑制1类HDAC外，SCFA还降低了PolyComb抑制剂复合物（PRC2）的3类HDAC SIRT1和亚基EZH2的表达，从而导致组蛋白过度乙酰化并同时减少抑制性组蛋白甲基化。基于这些结果，我们假设SCFA调节多个表观遗传调节剂，以促进HIV和疱疹病毒（例如KSHV）在口腔中的裂解复制。我们认为，SCFA还会影响宿主的细胞孔和传输途径，使细胞更允许病毒感染。对参与HIV转录激活的表观遗传调节剂的高吞吐量shRNA库筛选导致我们识别出许多表观遗传调节剂，这些调节剂似乎形成了多蛋白质沉默复合物。为了测试我们的假设，在AIM-I中，我们将执行蛋白质组学分析，以识别表观遗传沉默复合物的特定成分，并检查SCFAS对口腔上皮，内皮和CD4+ T-细胞中调节剂表达的影响。我们还将确认SCFA调节的表观遗传调节剂在我们的模型系统中HIV和KSHV的转录激活中的作用。先前的研究表明，细胞骨架肌动蛋白和微管的赖氨酸乙酰化在病毒进入和细胞内贩运方面起着重要作用。在AIM-II中，我们将检查是否以及如何 SCFA引起细胞骨架肌动蛋白和微管的赖氨酸乙酰化，从而增加病毒感染性，这是由于SCFAS抑制了1类HDAC和3级HDAC SIRT1。为了进一步检验我们的假设，在AIM-III中，我们将检查来自患有严重牙周疾病的HIV患者的口腔上皮和T细胞是否确实显示出AIM-I中检测到与SCFAS相关的表观遗传变化，以及表观遗传变化是否与患者的病毒复制和感染相关，是否与患者的病毒复制和感染相关。预计本研究将揭示新的和常见的机制，这些机制可以控制HIV患者口腔中HIV和疱疹病毒的复制和感染。 结果应为制定新的策略提供科学基础，以预防和治疗艾滋病毒/艾滋病患者的复发性艾滋病毒和疱疹病毒感染。