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

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，这将为我作为教员的研究奠定基础。这个项目将提供新技术（共聚焦显微镜、流式细胞术、凝胶移位）的培训，这些技术将适用于我作为独立研究者进行的研究。莱明斯博士的专业知识，以及微生物免疫学系的其他教职员工，对于学习成为一名成功的独立研究人员所必需的技能将是无价的。

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.