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（由申请方提供）：人乳头瘤病毒（HPV）由二十面体衣壳内相对较小的环状双链DNA基因组组成。在迄今发现的100多种HPV毒株中，约有40种基因型与生殖器疾病相关。HPV感染通常无症状，不会引起疾病。然而，如果病毒基因组维持在低水平（称为潜伏期）并逃避免疫识别，HPV可能导致异常细胞变化和严重疾病。感染生殖道的HPV可根据其致癌潜力和与癌症发展的相关性分为两组。低风险的HPV引起生殖器疣，很少与肿瘤发生有关。然而，一些高危HPV（如HPV 16）已被公认为宫颈癌的致病因子和其他几种上皮癌类型的辅因子。目前，这两种HPV疫苗只能针对有限的致癌HPV，并非所有国家都负担得起。在HPV进入过程中，硫酸乙酰肝素蛋白聚糖（HSPG）作为附着因子，以增加HPV感染的效率。与HSPG的结合导致病毒颗粒的构象变化和衣壳蛋白L2的裂解，随后是病毒粒子与次级未鉴定的进入受体的相互作用。HPV颗粒进入细胞主要通过内吞作用，在菌株和细胞类型依赖性的方式，和解体的病毒颗粒发生在内涵体在低pH值。细胞骨架已被建议参与HPV的细胞内运输。对HPV进入和传播机制的认识不足，制约了新疫苗和抗病毒药物的开发。全基因组siRNA筛选发现γ-分泌酶（J-sec）是HPV 16感染上皮细胞的重要因子，尽管其机制尚未得到很好的表征。本项目将研究J-sec及其底物在HPV 16感染人上皮细胞过程中的作用。本研究的第一个目的是通过新开发的技术特异性地测量病毒拆卸和病毒颗粒的定位来检查HPV 16感染过程中需要J-sec的关键步骤。J-sec活性丧失所抑制的步骤 将研究与各种细胞标记物的关系。本研究的第二部分将通过所提出的模型来研究J-sec的两种潜在底物在介导HPV 16进入中的作用。这一目标可能提供了一个机械的理解的绝对要求J-sec的HPV进入。总体而言，提出的目标将是有价值的理解HPV 16的进入过程和抗病毒治疗的新靶点的设计。