The essential role of gamma-secretase in human papillomavirus infection

γ-分泌酶在人乳头瘤病毒感染中的重要作用

• 批准号: 8783864
• 负责人: Wei Zhang
• 金额: $ 5.15万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8783864
• 关键词: Abnormal Cell; Accounting; Affect; Anogenital venereal warts; Antiviral Therapy; Binding; Binding Proteins; Biological Assay; Capsid; Capsid Proteins; Cell Line; Cell Nucleus; Cells; Cleaved cell; Complex; Country; Cytoskeleton; Developing Countries; Development; Disease; Endocytosis; Endosomes; Epithelial; Epithelial Cells; Flow Cytometry; Genes; Genital system; Genome; Genotype; HPV-High Risk; Heparan Sulfate Proteoglycan; Human; Human Papilloma Virus Vaccine; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 16; Immune; Immunofluorescence Immunologic; Infection; Integral Membrane Protein; Integration Host Factors; Intracellular Transport; L2 viral capsid protein; Laboratories; Lead; Ligation; Link; Low risk HPV; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Marketing; Measures; Mediating; Membrane; Modeling; Molecular; Mutation; Nucleocapsid; Oncogenic; Oncogenic Viruses; Pathway interactions; Peptide Hydrolases; Pharmaceutical Preparations; Process; Production; Proteins; Reporting; Research; Research Proposals; Role; Route; Scientist; Sexually Transmitted Diseases; Small Interfering RNA; Staging; Technology; Transmembrane Domain; United States; Vaccines; Viral; Virion; Virus; Virus-Cell Membrane Interaction; base; betacellulin; cancer type; cell type; cofactor; cost; design; ds-DNA; gamma secretase; genome-wide; high risk; improved; inhibitor/antagonist; novel; novel vaccines; particle; prevent; public health relevance; receptor; research study; sexually active; trafficking; tumorigenesis; uptake; vaccine development

DESCRIPTION (provided by applicant): Human papillomaviruses (HPVs) are composed of relatively small circular, double-stranded DNA genomes within icosahedral capsids. Among the over 100 HPV strains identified so far, approximately 40 genotypes are associated with genital diseases. HPV infection is generally asymptomatic and doesn't cause disease. However, if the virus genome is maintained at a low level termed latency and evades immune recognition, HPV may lead to abnormal cell changes and severe diseases. The HPVs that infect genital tract could be divided into two groups, based on their oncogenic potential and association with cancer development. The low-risk HPVs cause genital warts, and seldom linked to tumorigenesis. However, some high-risk HPVs (such as HPV16) have been well recognized as the causative agents of cervical cancer and as cofactors for several other epithelial cancer types. Currently, the two HPV vaccines can only target limited oncogenic HPVs and are not affordable in all the countries. During HPV entry, heparan sulfate proteoglycans (HSPGs) serve as the attachment factor to increase efficiency of HPV infection. Binding to HSPGs leads to the conformational change in the viral particles and cleavage of capsid protein L2, followed by the interaction of virion with a secondary unidentified entry receptor. HPV particles enter cells mainly through endocytosis in a strain and cell type dependent manner, and disassembly of virus particle occurs in endosomes at low pH. Cytoskeleton has been suggested to be involved in HPV intracellular trafficking. The poor understanding of entry and trafficking mechanism of HPV has restrained the development of new vaccines and anti-viral drugs. The genome-wide siRNA screen found gamma-secretase (J-sec) as an essential factor for HPV16 infection in epithelial cells, although the mechanism is not well characterized. This project will investigate the function of J-sec and its substrate(s) during the infectious route of HPV16 into human epithelial cells. The first aim of this study will examine the critical steps that require J-sec duing HPV16 infection by specifically measuring virus disassembly and the localization of virus particles with newly developed technologies. The steps that are inhibited by loss of J-sec activity will be investigated in relation to various cell markers. The second part of this research will examine the role of two potential substrates of J-sec in mediating HPV16 entry by proposed models. This aim may provide a mechanistic understanding of the absolute requirement of J-sec on HPV entry. Overall, the proposed aims will be valuable for the understanding of HPV16 entry process and the design of new targets for antiviral therapies.

描述（由申请人提供）：人乳头瘤病毒（HPV）由二十面体内衣壳内的相对较小的圆形，双链DNA基因组组成。在迄今为止鉴定出的100多个HPV菌株中，大约40种基因型与生殖器疾病有关。 HPV感染通常是无症状的，不会引起疾病。但是，如果病毒基因组保持在低水平的潜伏期并逃避免疫识别，则HPV可能导致异常的细胞变化和严重的疾病。感染生殖道的HPV可以根据其致癌潜力和与癌症发展的关联而分为两组。低风险的HPV会引起生殖器疣，很少与肿瘤发生有关。但是，一些高危HPV（例如HPV16）已被广泛认为是宫颈癌的病因，是其他几种上皮癌类型的辅助因子。目前，两种HPV疫苗只能针对有限的致癌HPV，并且在所有国家都不适合。在HPV进入期间，硫酸乙酰肝素蛋白聚糖（HSPG）是提高HPV感染效率的附着因子。与HSPGS的结合导致病毒颗粒的构象变化和衣壳蛋白L2的裂解，然后与二级未识别的入口受体相互作用。 HPV颗粒主要通过菌株和细胞类型依赖性方式进入细胞，而病毒颗粒的拆卸发生在低pH下的内体中。已建议细胞骨架参与HPV细胞内贩运。对HPV的进入和贩运机制的不良理解限制了新的疫苗和抗病毒药物的发展。全基因组siRNA筛查发现γ-分泌酶（J-SEC）是上皮细胞中HPV16感染的重要因素，尽管该机理的表征不佳。该项目将在HPV16传染性途径中研究J-SEC及其底物的功能。这项研究的第一个目的将检查需要J-SEC Duing HPV16感染的关键步骤，通过特异性测量病毒拆卸和使用新开发的技术的病毒颗粒定位。损失J-SEC活动所抑制的步骤 将根据各种细胞标记进行研究。这项研究的第二部分将研究J-SEC的两个潜在底物在通过建议模型中介导HPV16进入中的作用。这个目标可能提供对J-SEC在HPV进入的绝对要求的机械理解。总体而言，拟议的目标对于理解HPV16进入过程和设计抗病毒疗法的新目标将是有价值的。