Investigating novel host factors promoting human papillomavirus infection

研究促进人乳头瘤病毒感染的新宿主因素

• 批准号: 10544348
• 负责人: Mara Calypso Harwood
• 金额: $ 1.98万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10544348
• 关键词: Adult; Anogenital cancer; Antiviral Therapy; Binding; Biochemical; Biochemistry; Cadherins; Cell Adhesion; Cell Adhesion Molecules; Cell Nucleus; Cell membrane; Cell surface; Cells; Cellular biology; Complex; Cytosol; DNA Tumor Viruses; DNA Viruses; Data; Disease; Endocytosis; Endosomes; Event; Goals; Golgi Apparatus; Hand; Health; Human; Human Papilloma Virus Vaccine; Human Papilloma Virus-Related Malignant Neoplasm; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus HPV L1 protein; Image; Infection; Integral Membrane Protein; Integration Host Factors; Intracellular Transport; L2 viral capsid protein; Lysosomes; Malignant neoplasm of cervix uteri; Mass Spectrum Analysis; Mediating; Membrane; Microscopy; Mitosis; Modeling; Molecular Chaperones; Nuclear Envelope; Nucleoplasm; Pathway interactions; Population; Predisposition; Productivity; Proteins; Research; Role; Route; Series; Testing; Therapeutic; Time; Vesicle; Viral; Viral Proteins; Virus; Virus Diseases; combat; endosome membrane; experimental study; extracellular; gamma secretase; insight; malignant oropharynx neoplasm; novel; novel therapeutic intervention; particle; prophylactic; receptor; receptor mediated endocytosis; recruit; therapeutic target; trafficking; unvaccinated

Project Summary/Abstract: Human papillomavirus (HPV) infects nearly 80 million U.S. adults, and is the primary cause of cervical, anogenital, and oropharyngeal cancers. Unfortunately, HPV vaccines have been poorly utilized and provide no therapeutic value for existing infections, leaving much of the population susceptible to HPV-related cancers. To initiate infection, HPV binds to a series of extracellular receptors and is endocytosed by the host cell. HPV is then recruited to the host factor γ-secretase in the endosome by an unknown mechanism, enabling it to act as a chaperone to insert the viral capsid protein L2 across the endosomal membrane. This exposes L2 to the cytosol, which in turn recruits host factors that direct the virus along an infectious route through the Golgi apparatus and to the nucleus. Despite HPV’s significant impact on human health, there is limited understanding of its specific entry pathway through the host cell. Preliminary data suggest a role for the host p120-catenin (p120) in regulating the early trafficking steps of HPV along an infectious route. Our research group first identified an HPV-p120 interaction via mass-spectrometry, and we have since demonstrated that this interaction occurs at early infection time-points. Importantly, p120 is essential for productive HPV infection. Further experiments suggest that p120 is required for targeting HPV to γ- secretase for insertion of the viral capsid protein L2 across the endosomal membrane. However, as p120 is a cytosolic factor, it must be interacting with HPV via a transmembrane protein. Thus, we tested the role of cadherins – transmembrane cell adhesion molecules known to bind both p120 and γ-secretase – in HPV infection. Preliminary data show that in addition to binding HPV at early time-points, cadherins are also required for productive HPV infection. Together, these data suggest the cell-adhesion cadherin molecule, in conjunction with cytosolic p120, binds to HPV at the cell surface, and promotes endocytosis of the virus to the endosome. As p120 is known to deliver cadherin to γ-secretase, we further hypothesize that p120 targets the HPV-cadherin complex to endosome-localized γ-secretase so that the virus can reach γ- secretase for membrane insertion of the viral protein L2. To further test this hypothesis, we will elucidate the precise mechanism by which the cadherin-p120 complex promotes endocytosis of HPV to the endosome to initiate HPV infection (Aim 1), and how p120 then targets the virus to γ-secretase for membrane insertion (Aim 2) – a critical step of infection. The combined use of cell-based, biochemical, and infection studies will be employed to tackle the questions at hand. Insights gained through these studies should illuminate intrinsic intracellular transport mechanisms. More importantly, identifying the cellular factors mediating HPV infection may provide new therapeutic strategies to combat HPV infections.

项目概要/摘要： 人乳头瘤病毒（HPV）感染近8000万美国成年人，是宫颈癌的主要原因， 肛门生殖器和口咽癌。不幸的是，HPV疫苗的利用率很低， 对现有感染的治疗价值，使许多人容易患上HPV相关的癌症。到 在启动感染时，HPV与一系列细胞外受体结合并被宿主细胞内吞。HPV是 然后通过一种未知的机制招募到内体中的宿主因子γ-分泌酶，使其能够 作为伴侣蛋白将病毒衣壳蛋白L2插入内体膜。这暴露 L2进入细胞质，细胞质又招募宿主因子，引导病毒沿着感染途径通过细胞质。 高尔基体和细胞核。尽管HPV对人类健康有重大影响， 了解其通过宿主细胞的特定进入途径。 初步数据表明宿主p120-连环蛋白（p120）在调节HPV早期运输步骤中发挥作用 沿着一条传染途径。我们的研究小组首先通过质谱法确定了HPV-p120相互作用， 并且我们已经证明这种相互作用发生在早期感染时间点。重要的是，p120是 对生产性HPV感染至关重要。进一步的实验表明，p120是将HPV靶向γ- 分泌酶，用于将病毒衣壳蛋白L2插入穿过内体膜。因为p120是 作为细胞溶质因子，它必须通过跨膜蛋白与HPV相互作用。因此，我们测试了 钙粘蛋白-已知结合p120和γ-分泌酶的跨膜细胞粘附分子-在HPV 感染初步数据显示，除了在早期时间点结合HPV外，还需要钙粘蛋白 HPV感染的风险总之，这些数据表明，细胞粘附钙粘蛋白分子， 与胞质p120结合，在细胞表面与HPV结合，并促进病毒的内吞作用 到核内体。由于已知p120将钙粘蛋白递送至γ-分泌酶，我们进一步假设p120 将HPV-钙粘蛋白复合物靶向于内体定位的γ-分泌酶，使得病毒可以到达γ- 用于病毒蛋白L2的膜插入的分泌酶。为了进一步验证这一假设，我们将阐明 钙粘蛋白-p120复合物促进HPV内吞到内体的精确机制， 启动HPV感染（目的1），以及p120如何将病毒靶向γ-分泌酶进行膜插入（目的 2)- 感染的关键步骤。基于细胞的、生物化学的和感染研究的结合使用将是 用来解决手头的问题通过这些研究获得的见解应该阐明内在的 细胞内转运机制。更重要的是，鉴定介导HPV感染的细胞因子可能 提供新的治疗策略来对抗HPV感染。