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

基本信息

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
项目类别：
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

项目摘要

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肿瘤病毒领域的主要独立科学家。在获得独立奖的指导阶段，我将努力在HPV领域进一步建立我的出版记录，这将为我将作为教职员工做的研究奠定基础。该项目将提供新技术（共聚焦显微镜，流式细胞仪，凝胶转移）的培训，这些技术适用于我作为独立研究者进行的研究。 Laimins博士的专业知识以及微生物 - 免疫学系的其他教职员工在学习成为成功的独立研究人员的必要技能方面将是无价的。人乳头瘤病毒是小的DNA病毒，被认为是宫颈癌的病因。 HPV的传染性生命周期取决于细胞因子和上皮分化。分化触发了生命周期的生产阶段，其中包括病毒基因组扩增和病毒体的产生。该项目的长期目标是了解调节依赖分化病毒事件激活的机制。我最近证明，HPV-31在分化时刺激了低水平的caspase激活，这是线粒体凋亡途径的特征，对于通过裂解病毒复制蛋白E1的切割而进行了病毒基因组的扩增所必需的。基于以下发现，E6和E7可以刺激caspase激活，以及在几种HPV蛋白中鉴定caspase裂解位点的鉴定，我假设：HPV通过E6和E7诱导了caspase在区分中的caspase激活，以使细胞中的细胞中的病毒蛋白在激活后期病毒式事件中作用。在要在指导阶段启动的特定目标1中，我将通过检查E6和E7对线粒体膜完整性的影响以及在E6和E7中识别E6和E7中所需的E6的影响，从而确定HPV蛋白激活分化细胞中caspase的机制。在要在独立阶段开始的特定目标2中，我将通过使用体外和体内研究来确定HPV蛋白的caspase裂解如何在生命周期中有助于分化依赖性事件。描述E6和E7介导caspase在区分上皮中激活以导致病毒蛋白的裂解的途径将提供有关HPV如何调节凋亡机制的洞察力，不仅可以促进病毒生命周期，而且还可能转化。相关性：人乳头瘤病毒是宫颈癌的病因。了解病毒癌蛋白如何有助于调节病毒生命周期，最终将提供有关HPV如何引起疾病并提供潜在治疗靶标的洞察力。