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）：本研究的目的是研究牙周细菌代谢产物对宿主细胞表观遗传机制的调节，以及对增强艾滋病毒患者口腔中病毒复制和感染的影响。众所周知，HIV 患者慢性牙周炎的患病率和严重程度有所增加，并且更频繁地遭受疱疹病毒感染，例如单纯疱疹病毒 (HSV)、EB 病毒 (EBV) 和卡波西肉瘤相关疱疹病毒 (KSHV)。口腔也是潜在艾滋病毒感染的潜在储存库。先前的研究表明，牙周细菌会促进艾滋病毒和疱疹病毒的裂解性复制，从而导致牙周病复发 HIV 感染和口腔病变。厌氧革兰氏阴性菌产生各种短链脂肪酸 (SCFA)，可抑制 1/2 类组蛋白脱乙酰酶 (HDAC)，从而促进组蛋白过度乙酰化和病毒基因表达。然而，短链脂肪酸是否影响其他表观遗传调节因子以及它们是否也会增强病毒感染尚未得到研究，仍有待确定。我们的初步研究数据表明，短链脂肪酸不仅诱导 HIV 和 KSHV 的裂解性复制，而且还增加了这两种病毒在各自靶细胞中的感染率。除了抑制 1/2 类 HDAC 外，SCFA 还下调 3 类 HDAC SIRT1 和多梳抑制复合物 (PRC2) 亚基 EZH2 的表达，同时导致组蛋白过度乙酰化和抑制性组蛋白甲基化减少。基于这些结果，我们假设 SCFA 调节多个表观遗传调节因子，以促进 HIV 和疱疹病毒（例如 KSHV）在口腔中的裂解性复制。我们相信 SCFA 还会影响宿主细胞骨架和运输途径，使细胞更容易受到病毒感染。通过高通量 shRNA 文库筛选参与 HIV 转录激活的表观遗传调节因子，我们鉴定出了许多似乎形成多蛋白沉默复合物的表观遗传调节因子。为了检验我们的假设，在 AIM-I 中，我们将进行蛋白质组分析，以确定表观遗传沉默复合物的特定成分，并检查 SCFA 对口腔上皮细胞、内皮细胞和 CD4+ T 细胞中调节因子表达的影响。我们还将确认 SCFA 调节的表观遗传调节因子在我们的模型系统中 HIV 和 KSHV 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运输中发挥重要作用。在 AIM-II 中，我们将研究是否以及如何 SCFA 会导致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 SCFA 抑制 1/2 类 HDAC 和 3 类 HDAC SIRT1 的结果。为了进一步检验我们的假设，在 AIM-III 中，我们将检查患有严重牙周病的 HIV 患者的口腔上皮和 T 细胞是否确实表现出 AIM-I 中检测到的 SCFA 相关表观遗传变化，以及表观遗传变化是否与病毒复制相关通过进行横断面和纵向人体研究来确定患者的感染情况。预计本研究将揭示控制艾滋病毒和疱疹病毒在艾滋病毒患者口腔中复制和感染的新颖且常见的机制。 研究结果将为制定同时预防和治疗艾滋病毒/艾滋病患者复发性艾滋病毒和疱疹病毒感染的新策略提供科学依据。

项目成果

Short-chain fatty acids from periodontal pathogens suppress histone deacetylases, EZH2, and SUV39H1 to promote Kaposi's sarcoma-associated herpesvirus replication.

来自牙周病原体的短链脂肪酸抑制组蛋白脱乙酰酶、EZH2 和 SUV39H1，从而促进卡波西肉瘤相关疱疹病毒的复制。

• 发表时间: 2014-04
• 作者: Yu, Xiaolan;Shahir, Abdel;Sha, Jingfeng;Feng, Zhimin;Eapen, Betty;Nithianantham, Stanley;Das, Biswajit;Karn, Jonathan;Weinberg, Aaron;Bissada, Nabil F;Ye, Fengchun
• 通讯作者: Ye, Fengchun