权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Mechanisms of human papillomavirus entry

人乳头瘤病毒的侵入机制

基本信息

批准号：
10675774
负责人：
Daniel C. Dimaio
金额：
$ 98.49万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-17 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10675774
关键词：
Adopted Amaze Anogenital venereal warts Binding Binding Sites Biology Cancer Etiology Capsid Proteins Cell Nucleus Cell physiology Cells Cellular biology Cytoplasm Funding Future Genetic Grant HIV HPV analysis Human Human Papilloma Virus-Related Malignant Neoplasm Human Papillomavirus Human papilloma virus infection Infection Integral Membrane Protein Journals L2 viral capsid protein Malignant Neoplasms Maps Membrane Minority Groups National Cancer Institute Oncogenic Viruses Papilloma Papillomavirus Pathway interactions Penetration Peptides Persons Phenotype Proteins Public Health Publishing Role Seminal Sorting System Testing Time Vaccination Vaccines Virion Virus Virus Diseases anticancer research design experimental study genetic analysis genetic approach innovation insight mutant novel novel strategies pathogen prevent protein aminoacid sequence receptor retrograde transport trafficking tumor unvaccinated vaccine acceptance virology virome

项目摘要

This is a revised application for an R35 grant from the National Cancer Institute entitled, “Mechanisms of human papillomavirus entry.” HPV is responsible for 5% of human cancer, millions of cases of genital warts, and countless cases of other types of papillomas (most caused by non-vaccine HPV types). Despite the existence of effective vaccines, HPV infection and the cancers it causes will remain a major public health problem for decades because vaccine uptake is poor and vaccination does not clear established infections. There are no specific treatments for HPV or HPV-associated cancers. The DiMaio lab has received more than 35 years of continuous funding from the NCI, during which time it has made seminal contributions to tumor virology. We recently discovered that HPV traffics via the retrograde transport pathway during infection, showed that retromer is required for sorting of the incoming virus particle into this pathway and that HPV is a novel type of retromer cargo, and discovered a cell-penetrating peptide (CPP) that drives the HPV L2 capsid protein into the cytoplasm to engage retromer. This is the first example where either retromer or a CPP has been shown to play a role in virus entry. These results have been paradigm-shifting in the field and have been published since 2013 in Cell, PLoS Pathogens, mBio, PNAS, and Journal of Cell Biology, and they have fundamentally changed our understanding of HPV entry and the role of retromer and CPPs in biology. Here, we will discover how HPV accomplishes these amazing feats. We will determine the requirements for L2 membrane protrusion and establish how sequences flanking the core CPP modulate its activity. We will test whether the abundance of CPP sequences in the extant papillomavirus virome reflects their membrane- penetrating activity, determine whether L2 truly adopts a transmembrane existence, and map L2 segments exposed in the cytoplasm. We have designed an innovative new functional genetics approach and used it to isolate artificial small transmembrane proteins that inhibit HPV entry, and we will use these artificial proteins to identify new HPV entry factors and dissect their role in HPV entry. We will exploit our understanding of HPV entry to design inhibitory peptides that harness the membrane-penetrating activity of CPPs to deliver the retromer binding site into the cytoplasm to compete for binding with incoming HPV, validating an entirely new approach to prevent virus infections. We will develop the first genetic system to select HPV mutants with informative phenotypes, attempt to identify the HPV entry receptor, and extend these studies to additional tumor viruses and HIV. These experiments will elucidate important aspects of the mechanisms of HPV entry, validate new approaches to prevent and treat HPV infection, and revolutionize genetic analysis of HPV. Critically, our studies will provide new insights into fundamental cell biology. If this proposal is funded, we will continue to make novel and important contributions to virology and cancer research for many years.

这是一份来自国家癌症研究所的修订后的R35拨款申请，标题为人乳头瘤病毒进入。人类乳头状瘤病毒导致了5%的人类癌症，数百万生殖器疣病例，以及无数其他类型的乳头状瘤(大多数由非疫苗类型的HPV引起)。尽管有效疫苗的存在、HPV感染及其引起的癌症仍将是一个主要的公共卫生问题几十年来一直是个问题，因为疫苗接种率很低，而且疫苗接种不能清除已确定的感染。目前还没有针对HPV或HPV相关癌症的特效治疗方法。迪马约实验室已经收到了超过 NCI连续资助了35年，在此期间它为肿瘤做出了开创性的贡献病毒学。我们最近发现，在感染期间，HPV通过逆行运输途径进行传播，表明需要逆转聚体来将传入的病毒颗粒分类到这一途径中，并且HPV是一种新型逆转录病毒载体，并发现了驱动HPV L2衣壳的细胞穿透肽(CPP) 蛋白质进入细胞质进行逆转录。这是第一个反转录或CPP具有已被证明在病毒入侵过程中起到了作用。这些结果在该领域发生了范式转变，并已自2013年以来发表在《细胞》、《公共科学图书馆·病原体》、《mBio》、《PNAS》和《细胞生物学杂志》上，他们已经从根本上改变了我们对HPV进入以及逆转录和CPP在生物学中的作用的理解。这里, 我们将发现HPV是如何完成这些惊人的壮举的。我们将确定L2的要求膜突起，并确定核心CPP两侧的序列如何调节其活性。我们将测试现存的乳头瘤病毒中CPP序列的丰度是否反映了它们的膜- 穿透活性，确定L2是否真的采用跨膜存在，并映射L2片段暴露在细胞质中。我们设计了一种创新的新功能遗传学方法，并将其用于分离抑制HPV进入的人工小跨膜蛋白，我们将使用这些人工蛋白来确定新的HPV进入因素，并剖析它们在HPV进入中的作用。我们将充分利用我们对HPV的理解进入设计抑制肽，利用CPPs的膜穿透活性来传递将结合部位反转录到细胞质中，与传入的HPV竞争结合，验证了一种全新的预防病毒感染的方法。我们将开发第一个用于选择HPV突变体的遗传系统信息性表型，试图确定HPV进入受体，并将这些研究扩展到其他肿瘤病毒和艾滋病毒。这些实验将阐明HPV进入机制的重要方面，验证预防和治疗HPV感染的新方法，并彻底改变HPV的基因分析。重要的是，我们的研究将为基础细胞生物学提供新的见解。如果这项提案获得资金，我们将多年来继续为病毒学和癌症研究做出新的重要贡献。