Mechanisms of Interferon Regulatory Factor Dysfunction by Cutaneous Papillomaviruses

皮肤乳头瘤病毒干扰素调节因子功能障碍的机制

• 批准号: 10217281
• 负责人: JENNIFER A LUFF
• 金额: $ 12.93万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10217281
• 关键词: Address; Advanced Malignant Neoplasm; Animal Model; Animals; Antiviral Agents; Antiviral Response; Binding; Bioinformatics; Biology; Bypass; Canis familiaris; Cell Differentiation process; Cells; Co-Immunoprecipitations; Complement; Confocal Microscopy; Cutaneous; Data; Development; Faculty; Foundations; Frequencies; Functional disorder; Future; Gene Expression; Genes; Genetic; Genetic Diseases; Goals; HIV; HIV Infections; Health; Human; Human Papillomavirus; Immune; Immunocompromised Host; Immunologic Deficiency Syndromes; Individual; Infection; Infectious Skin Diseases; Interferon-beta; Interferons; Investigation; Malignant neoplasm of cervix uteri; Mass Spectrum Analysis; Mediating; Mentors; Modeling; Mucous Membrane; Oncogenes; Organ Transplantation; Papillomavirus; Papillomavirus Infections; Patients; Proteomics; Regulation; Repression; Research; Risk; Role; Scientist; Skin; Skin Carcinoma; Stimulus; Sum; Testing; Therapeutic; Thromboplastin; Tumor Suppressor Proteins; Virus Diseases; Work; cell type; chromatin immunoprecipitation; cofactor; human disease; inducible gene expression; keratinocyte; kidney cell; new therapeutic target; novel therapeutics; response; skills; targeted treatment; therapeutic target; treatment response; tumor; tumor progression; virtual

Project Summary/ Abstract Papillomavirus (PV) infections significantly impact human health. Mucosal PVs cause virtually all cases of human cervical cancer, while cutaneous PVs cause severe skin infections and non-melanoma skin cancers in patients with immunodeficiencies resulting from e.g. HIV infection, organ transplants, or genetic disorders. Treatments for cutaneous PVs are limited and often ineffective. Thus, there is a critical need for development of novel therapeutics directed at keratinocytes, the target cell of PV infections. Keratinocytes have the potential to mount an antiviral response. Viral infection activates interferon regulator factors (IRFs), which upregulate interferons (IFN) and IFN-stimulated antiviral genes. IRFs and IFNs are also tumor-suppressing, making them particularly attractive as therapeutic targets for PV infections. However, both mucosal and cutaneous PVs can repress IRF function and IFN expression. This repression is mediated by the E6 and E7 oncogenes of mucosal PVs. While limited studies suggest that mucosal and cutaneous PVs differ in their E6 and E7 functions and that IRF regulation of the IFN response in keratinocytes differs from that in other cell types, there are many more questions than answers. Like humans, cutaneous PV infections occur in dogs and are more prevalent in immunodeficient animals. Thus, the dog offers a natural spontaneous animal model for investigation of cutaneous PV infections and for testing novel therapeutics. Indeed, our preliminary data demonstrate that canine cutaneous PV E6 and E7 differentially disrupt IFN and IFN-stimulated gene expression in canine keratinocytes. The long-range goal of our work is to develop novel therapeutics targeting IRFs and the IFN response for treatment of cutaneous PV infections in immunodeficient patients, using a canine model of PV infection. We hypothesize here that E6 and E7 from cutaneous PVs enhance viral infection by inhibiting IRF function in keratinocytes using mechanisms unique to cutaneous PVs. In Aim 1, we will inhibit expression of IRFs to determine their impact on cutaneous PV infection and on constitutive and inducible expression of IFNs and IFN-stimulated genes in keratinocytes. In Aims 2 and 3, we will determine the mechanisms for cutaneous PV E6- and E7-mediated disruption of IFNs and IFN-stimulated gene expression in keratinocytes. Specifically, we will use mass spectrometry to identify E6 and E7 binding partners that modulate IRF expression and/or function, and then identify mechanisms for E6 and E7 effects on these binding partners. Using human PV and keratinocytes will address the human disease; using canine PV and keratinocytes will advance the dog model. Results will lay the foundation for development of novel therapeutics that would by-pass E6 and E7 deleterious effects. The mentoring team includes faculty with extensive expertise in keratinocyte biology, IRF and IFN biology in differentiated cells, PV biology, and mass spectrometry proteomics and bioinformatics. In sum, the proposed research will address a critical issue in human and animal health, capitalize on the diverse skill sets of the interdisciplinary mentoring team, and enhance my maturation as an independent clinician scientist.

项目概要/摘要 乳头瘤病毒（PV）感染严重影响人类健康。几乎所有病例均由粘膜 PV 引起 人类宫颈癌，而皮肤 PV 会导致严重的皮肤感染和非黑色素瘤皮肤癌 患有免疫缺陷的患者，例如HIV 感染、器官移植或遗传性疾病。 皮肤PV的治疗方法有限且常常无效。因此，迫切需要发展 针对角化细胞（PV 感染的靶细胞）的新型疗法。角质形成细胞具有潜力 发起抗病毒反应。病毒感染会激活干扰素调节因子 (IRF)，从而上调 干扰素 (IFN) 和 IFN 刺激的抗病毒基因。 IRF 和 IFN 还具有肿瘤抑制作用，因此 作为真性红斑狼疮感染的治疗靶点特别有吸引力。然而，粘膜和皮肤 PV 都可以 抑制 IRF 功能和 IFN 表达。这种抑制是由粘膜的 E6 和 E7 癌基因介导的。 光伏发电。虽然有限的研究表明粘膜和皮肤 PV 的 E6 和 E7 功能不同，并且 IRF 对角质形成细胞中 IFN 反应的调节与其他细胞类型不同，还有更多 问题多于答案。与人类一样，皮肤PV感染也发生在狗身上，并且在以下人群中更为普遍： 免疫缺陷动物。因此，狗提供了一种自然自发的动物模型来研究 皮肤PV感染和测试新疗法。事实上，我们的初步数据表明 犬皮肤 PV E6 和 E7 差异性破坏犬体内 IFN 和 IFN 刺激的基因表达 角质形成细胞。我们工作的长期目标是开发针对 IRF 和 IFN 的新型疗法 使用犬PV模型治疗免疫缺陷患者皮肤PV感染的反应 感染。我们假设来自皮肤PV的E6和E7通过抑制IRF来增强病毒感染 使用皮肤 PV 独特的机制在角质形成细胞中发挥作用。在目标 1 中，我们将抑制 IRF 以确定其对皮肤 PV 感染以及 IFN 组成型和诱导型表达的影响 和角质形成细胞中干扰素刺激的基因。在目标 2 和 3 中，我们将确定皮肤的机制 PV E6 和 E7 介导的 IFN 破坏以及角质形成细胞中 IFN 刺激的基因表达。具体来说， 我们将使用质谱法来鉴定调节 IRF 表达和/或 功能，然后确定 E6 和 E7 对这些结合伙伴的影响机制。利用人类PV和 角质形成细胞将解决人类疾病；使用犬 PV 和角质形成细胞将改进狗模型。 结果将为开发绕过 E6 和 E7 有害物质的新型疗法奠定基础 影响。指导团队包括在角质形成细胞生物学、IRF 和 IFN 方面拥有丰富专业知识的教师 分化细胞生物学、光伏生物学、质谱蛋白质组学和生物信息学。总而言之， 拟议的研究将解决人类和动物健康中的一个关键问题，并利用多样化的技能 跨学科指导团队的帮助，并提高我作为一名独立临床科学家的成熟度。