Epigenetic Regulation of HSV Infection of Oral Cells

描述(由申请人提供)：表观遗传机制控制许多正常和疾病状态的过程。在这项建议中，我们将使用单纯疱疹病毒1型(HSV-1)作为在正常和疾病口腔上皮细胞中操作的表观遗传学机制的探针。我们的具体目标是：1.明确HSV-1基因在正常口腔角质形成细胞中表达的表观遗传调控机制。我们将首先定义永生化的正常口腔角质形成细胞(NOK细胞)中HSV基因表达的表观遗传调控的基本机制，定义口腔角质形成细胞中染色质和异染色质与病毒裂解基因相关的动力学，定义正负向调节NOK细胞中病毒基因表达的宿主染色质因子，并测试靶向染色质因子的药物与传统疱疹病毒药物的协同作用能力。2.验证HSV-1感染可以作为不同口腔疾病状态表观遗传机制的探针的假设。HIV感染口腔上皮细胞的表观遗传机制在抗逆转录病毒治疗中发生改变 细胞，以及肿瘤细胞中。我们发现HSV-1感染可以作为骨肉瘤细胞的探针，以确定在这些细胞中起作用的新的表观遗传机制。因此，我们将测试HSV-1感染是否可以在患病的口腔细胞中识别出改变的表观遗传机制。在这一目标中，我们将确定HIV+/抗逆转录病毒治疗的个体与HIV-个体的口腔上皮细胞以及HIV蛋白酶抑制剂治疗的口腔上皮细胞中的表观遗传学机制，并定义在口腔肿瘤细胞中的表观遗传学机制。3.明确口腔疾病状态对HSV DNA IFI16核感觉的影响。我们已经证明，在包括口腔角质形成细胞在内的正常人细胞中，核IFI16DNA传感器是诱导干扰素和干扰素刺激的基因以及抑制HSV IE宿主沉默所必需的。在这一目标中，我们将确定在接受HIV+/抗逆转录病毒治疗的口腔角质形成细胞与接受HIV+/抗逆转录病毒治疗的患者的口腔角质形成细胞以及使用HIV蛋白酶抑制剂治疗的口腔上皮细胞中，HSV DNA的IFI16感觉是否发生了变化。在这个目标中，我们将确定口腔肿瘤细胞中HSV-1DNA的IFI16敏感性是否发生改变。这些关于HSV-1基因表达的表观遗传调控的研究有可能确定口腔上皮细胞中病毒感染和疾病的新机制和新的抗病毒策略。此外，这些研究将为确定口腔癌的致癌机制和治疗方法提供新的机制。

DESCRIPTION (provided by applicant): Epigenetic mechanisms control many normal and diseased state processes. In this proposal we will use herpes simplex virus 1 (HSV-1) as a probe of the epigenetic mechanisms operative in normal and diseased oral epithelial cells. Our specific aims are: 1. To define the mechanisms of epigenetic regulation of HSV-1 gene expression in normal oral keratinocytes. We will first define the baseline mechanisms of epigenetic regulation of HSV gene expression in immortalized normal human oral keratinocytes (NOK cells), define the kinetics of chromatin and heterochromatin association with viral lytic genes in oral keratinocytes, define the host chromatin factors that positively and negatively regulate viral gene expression in NOK cells, and test the ability of drugs that target chromatin factors to synergize with traditional herpesviral drugs. 2. Test the hypothesis that HSV-1 infection can be used as a probe of epigenetic mechanisms in different oral disease states. Epigenetic mechanisms are altered in HIV-infected oral epithelial cells, in anti-retroviral treated cells, and in tumor cells. We have found that HSV-1 infection can be used as a probe of osteosarcoma cells to identify novel epigenetic mechanisms that are operative in these cells. Therefore, we will test if HSV-1 infection can identify altered epigenetic mechanisms in diseased oral cells. In this Aim we will define the epigenetic mechanisms in oral epithelial cells from HIV+/anti-retroviral treated individuals versus HIV- individuals and in oral epithelial cells treatd with HIV protease inhibitors, and define the epigenetic mechanisms operative in oral tumor cells. 3. To define the effect of oral disease states on IFI16 nuclear sensing of HSV DNA. We have shown that in normal human cells including oral keratinocytes that the nuclear IFI16 DNA sensor is required for induction of IFN-¿ and interferon-stimulated genes and for host silencing of HSV IE. In this Aim we will determine if IFI16 sensing of HSV DNA is altered in oral keratinocytes from HIV+/anti-retroviral treated individuals versus HIV- individuals, and in oral epithelial cells treated with HIV protease inhibitors. In this aim we will determine if IFI16 sensig of HSV-1 DNA is altered in oral tumor cells. These studies on epigenetic regulation of HSV-1 gene expression have the potential of defining new mechanisms of viral infection and disease in oral epithelial cells and new antiviral strategies. Furthermore, the studies will provide new mechanisms for identifying oncogenic mechanisms and therapies for oral cancers.