The Role of Caspase Activation in the Differentiation-Dependent Life Cycle of HPV

Caspase 激活在 HPV 分化依赖性生命周期中的作用

• 批准号: 7574657
• 负责人: CARY A MOODY
• 金额: $ 9万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-20 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7574657
• 关键词: Address; Affect; Amino Acid Motifs; Apoptotic; Award; Caspase; Cell Cycle; Cells; Characteristics; Cleaved cell; Confocal Microscopy; DNA Tumor Viruses; DNA Viruses; Development; Disease; Environment; Epithelial; Epithelial Cells; Epithelium; Event; Faculty; Fellowship; Flow Cytometry; Gel; Gene Expression; Genital system; Genome; Goals; Human Papilloma Virus Vaccine; Human Papillomavirus; Human papilloma virus 31; Human papillomavirus 16 E1 protein; Immunology; In Vitro; Lead; Learning; Life Cycle Stages; Malignant neoplasm of cervix uteri; Mediating; Mentors; Microbiology; Mitochondria; Mucous Membrane; Normal Cell; Oncogene Proteins; Pathway interactions; Phase; Postdoctoral Fellow; Production; Proteins; Publications; Regulation; Research; Research Personnel; Risk; Role; Scientist; Site; TP53 gene; Techniques; Therapeutic; Training; Viral; Viral Genome; Viral Pathogenesis; Viral Proteins; Virion; Work; base; career; caspase-9; in vivo; insight; keratinocyte; member; mitochondrial membrane; novel; novel therapeutics; prophylactic; protein function; skills; therapeutic target

DESCRIPTION (provided by applicant): My career goal is to become a leading independent scientist in the DNA tumor virus field. During the mentored phase of the Pathway to Independence award, I will work to further establish my publication record in the HPV field which will lay the groundwork for the research I will do as a faculty member. This project will provide training with new techniques (confocal microscopy, flow cytometry, gel shifts) that will be applicable to the research I perform as an independent investigator. The expertise of Dr. Laimins, as well as other faculty members in the Microbiology-Immunology Department, will be invaluable in learning the necessary skills to become a successful independent researcher. Human papillomaviruses are small, DNA viruses that are considered the etiological agents of cervical cancer. The infectious life cycle of HPV is dependent on cellular factors and epithelial differentiation. Differentiation triggers the productive phase of the life cycle, which includes viral genome amplification and virion production. The long-term goal of this project is to understand the mechanisms that regulate the activation of differentiation-dependent viral events. I recently demonstrated that HPV-31 stimulates a low level of caspase activation upon differentiation that is characteristic of the mitochondrial apoptotic pathway and necessary for amplification of viral genomes through cleavage of the viral replication protein E1. Based on the finding that caspase activation could be stimulated by both E6 and E7, and the identification of caspase cleavage sites in several HPV proteins, I hypothesize that: HPV, through E6 and E7, induces caspase activation in differentiating cells to modify viral protein function as a mechanism to activate late viral events. In specific Aim 1, to be initiated during the mentored phase, I will identify the mechanism(s) by which HPV proteins activates caspases in differentiating cells by examining the effect of E6 and E7 on mitochondrial membrane integrity and identifying domains in E6 and E7 that are necessary for caspase activation. In Specific Aim 2, to be initiated during the independent phase, I will determine how caspase cleavage of HPV proteins contributes to differentiation-dependent events in the life cycle by using in vitro and in vivo studies. Delineating the pathway by which E6 and E7 mediate caspase activation in differentiating epithelia to result in cleavage of viral proteins will offer insight as to how HPV modulates apoptotic machinery to facilitate not only the viral life cycle, but possibly transformation as well. RELEVANCE: Human papillomaviruses are the causative agents of cervical cancer. Understanding how the viral oncoproteins contribute to regulation of the viral life cycle will ultimately offer insight into how HPV causes disease and provide potential therapeutic targets.

描述（由申请人提供）：我的职业目标是成为DNA肿瘤病毒领域领先的独立科学家。在独立之路奖的指导阶段，我将努力进一步建立我在 HPV 领域的发表记录，这将为我作为教员进行的研究奠定基础。该项目将提供新技术（共焦显微镜、流式细胞术、凝胶位移）的培训，这些技术将适用于我作为独立研究者进行的研究。莱明斯博士以及微生物学-免疫学系其他教员的专业知识对于学习成为一名成功的独立研究员所需的技能至关重要。 人乳头瘤病毒是小型 DNA 病毒，被认为是宫颈癌的病原体。 HPV 的感染生命周期取决于细胞因素和上皮分化。分化触发生命周期的生产阶段，包括病毒基因组扩增和病毒粒子产生。该项目的长期目标是了解调节分化依赖性病毒事件激活的机制。我最近证明，HPV-31 在分化时会刺激低水平的半胱天冬酶激活，这是线粒体凋亡途径的特征，也是通过切割病毒复制蛋白 E1 来扩增病毒基因组所必需的。基于 E6 和 E7 均可刺激 caspase 激活的发现，以及对几种 HPV 蛋白中 caspase 裂解位点的鉴定，我推测：HPV 通过 E6 和 E7 诱导分化细胞中的 caspase 激活，从而改变病毒蛋白功能，作为激活晚期病毒事件的机制。在指导阶段启动的具体目标 1 中，我将通过检查 E6 和 E7 对线粒体膜完整性的影响并识别 E6 和 E7 中 caspase 激活所需的结构域，确定 HPV 蛋白激活分化细胞中 caspase 的机制。在独立阶段启动的具体目标 2 中，我将通过体外和体内研究确定 HPV 蛋白的半胱天冬酶裂解如何促进生命周期中的分化依赖性事件。描述 E6 和 E7 在分化上皮细胞中介导 caspase 激活以导致病毒蛋白裂解的途径，将有助于了解 HPV 如何调节细胞凋亡机制，不仅促进病毒生命周期，而且还可能促进病毒转化。 相关性：人乳头瘤病毒是宫颈癌的病原体。了解病毒癌蛋白如何调节病毒生命周期最终将有助于深入了解 HPV 如何引起疾病并提供潜在的治疗靶点。