Mechanism of endosomal membrane penetration by Human Papillomavirus 16

人乳头瘤病毒16穿透内体膜的机制

• 批准号: 8978493
• 负责人: Kylia Goodner-Bingham
• 金额: $ 3.53万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-01 至 2016-01-27
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8978493
• 关键词: Acids; Address; Affect; Amino Acid Sequence; Biochemical; Biological Assay; C-terminal; Cancer Etiology; Capsid Proteins; Cells; Centrifugation; Clinical; Complex; Cytoplasm; Developing Countries; Development; Disease; Drug Targeting; Endosomes; Fluorescence; Force of Gravity; Goals; Golgi Apparatus; Hour; Human; Human Papilloma Virus Vaccine; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 16; Immunoblotting; Immunofluorescence Immunologic; Incidence; Individual; Infection; Infection prevention; Knowledge; L2 viral capsid protein; Label; Life Cycle Stages; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Measures; Membrane; Microscopy; Minor; Mutation; Pathway interactions; Penetration; Peptide Hydrolases; Peptide Sequence Determination; Peptides; Prevalence; Prevention; Process; Reporting; Research; Specificity; Structure; Techniques; Therapeutic; Thrombin; Vaccines; Vesicle; Viral; Virus; Virus Diseases; Woman; cost; cost effective; cost effectiveness; insight; malignant oropharynx neoplasm; men; mutant; novel; polypeptide C; prevent; prophylactic; protein aminoacid sequence; public health relevance; streptolysin O; trafficking

 DESCRIPTION (provided by applicant): Up to 99% of cervical cancer and 90% of oropharyngeal cancer cases are caused by Human Papillomavirus (HPV) infections. Although two HPV vaccines are in clinical use for the prevention of HPV- associated-cancers, there are limitations in the scope and worldwide cost-effectiveness of the current vaccines. In particular, while the vaccines may be prophylactic, they are not likely to be therapeutic. Moreover, the vaccines are only effective against four HPV types. Therefore, it is important to develop novel anti-viral therapies that can augment the efficacy of the current available therapies to prevent and treat HPV infection. One strategy is to develop drugs that target specific steps of the HPV life cycle. However, knowledge of the HPV infectious trafficking pathway is incomplete, and further research is required to more clearly define this pathway in order to identify novel drug targets. Evidence suggests that after the virus enters the cell, it traffics from the early endosom to Golgi. In support of this, it was recently shown that the retromer, a major cellular trafficking complex involved in endosome to Golgi transport, is crucial for HPV infection. Specifically, the viral minor capsid protein L2, within an endosomal vesicle, is able to directly interact with the cytoplasmic retromer complex in order to traffic from the endosome to the Golgi. Exactly how virus located within the endosome is able to recognize the retromer complex in the cytoplasm is currently unknown. This proposal seeks to address the mechanism by which HPV capsid proteins penetrate the endosomal membrane for gaining access to the retromer. To address this goal, I will identify HPV mutants that accumulate in the endosome and thereby determine the viral capsid protein sequences necessary for endosomal membrane penetration. Microscopy and biochemical techniques will be used to assess capsid protein penetration of the endosome. Overall, this study should provide further insight into the HPV trafficking pathway and thus aid in the development of anti-viral therapeutics for the prevention of HPV-associated cancers.

 描述（由申请人提供）：高达99%的宫颈癌和90%的口咽癌病例是由人乳头瘤病毒（HPV）感染引起的。尽管两种HPV疫苗在临床上用于预防HPV相关癌症，但目前疫苗的范围和全球成本效益存在局限性。特别是，虽然疫苗可能是预防性的，但它们不太可能是治疗性的。此外，疫苗仅对四种HPV类型有效。因此，重要的是开发新的抗病毒疗法，其可以增强当前可用疗法的功效以预防和治疗HPV感染。一种策略是开发针对HPV生命周期特定步骤的药物。然而，HPV感染性运输途径的知识是不完整的，需要进一步的研究来更清楚地定义这一途径，以确定新的药物靶点。有证据表明，病毒进入细胞后，它从早期的内体运输到高尔基体。为了支持这一点，最近有研究表明，逆转录病毒（retromer）是一种主要的细胞贩运 参与内体到高尔基体转运的复合体，对于HPV感染至关重要。具体地，病毒次要衣壳蛋白L2在内体囊泡内能够直接与细胞质逆转录聚合物复合物相互作用，以便从内体运输到高尔基体。目前还不清楚位于内体中的病毒如何能够识别细胞质中的逆转录复合物。该提案旨在解决HPV衣壳蛋白穿透内体膜以接近逆转录酶的机制。为了实现这一目标，我将鉴定在内体中积累的HPV突变体，从而确定内体膜穿透所需的病毒衣壳蛋白序列。将使用显微镜和生物化学技术来评估衣壳蛋白对内体的渗透。总的来说，这项研究应该为HPV运输途径提供进一步的见解，从而有助于 开发用于预防HPV相关癌症的抗病毒疗法。