Mechanisms of human papillomavirus entry

人乳头瘤病毒的侵入机制

• 批准号: 10042803
• 负责人: Daniel C. Dimaio
• 金额: $ 100.5万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 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/10042803
• 关键词: Adopted; Amaze; Anogenital venereal warts; Binding; Binding Sites; Biology; Cancer Etiology; Cell Nucleus; Cell physiology; Cells; Cellular biology; Cytoplasm; Funding; Future; Genetic; Grant; HIV; 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; National Cancer Institute; Oncogenic Viruses; Papilloma; Papillomavirus; Pathway interactions; Peptides; Phenotype; Play; Population; Proteins; Public Health; Publishing; Role; Seminal; Sorting - Cell Movement; 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; uptake; virology; virome

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.

这是国家癌症研究所的R35赠款的修订申请，题为“ 人类乳头瘤病毒的进入。 以及其他类型的乳头瘤（最多是由非疫苗HPV类型引起的）的无数情况。尽管有 存在有效的疫苗，HPV感染及其引起的癌症将仍然是主要的公共卫生 几十年来的问题是因为疫苗的摄取量很差，疫苗无法清除已建立的感染。 HPV或HPV相关癌症没有特定的治疗方法。 Dimaio实验室收到的超过 NCI的35年持续资金，在此期间，它已使肿瘤半分配 病毒学。我们最近发现，在感染期间，HPV通过逆行运输途径的流量， 表明将传入的病毒颗粒分类到该途径中是必需的，并且HPV是一个 新型逆转录货物的新型类型，并发现了驱动HPV L2 Capsid的细胞穿透肽（CPP） 蛋白质进入细胞质以吸收逆转录剂。这是撤回器或CPP具有的第一个示例 他们被证明在病毒进入中发挥作用。这些结果是在现场的范式转移，已经是 自2013年以来发表于细胞，PLOS病原体，MBIO，PNA和细胞生物学杂志，它们具有 基本上改变了我们对HPV进入的理解以及逆转录和CPP在生物学中的作用。这里， 我们将发现HPV如何完成这些惊人的壮举。我们将确定L2的要求 膜突出并确定核心CPP侧面如何调节其活性的序列。我们将测试 现存乳头瘤病毒病毒瘤中CPP序列的抽象是否反映其膜 穿透活动，确定L2是否真正适应了跨膜的存在，并映射L2段 暴露于细胞质中。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进入的人造人造小跨膜蛋白，我们将使用这些人造蛋白 确定新的HPV进入因素，并剖析其在HPV进入中的作用。我们将利用我们对HPV的理解 进入设计抑制性辣椒，以利用CPP的膜渗透活性以提供 逆转录位点进入细胞质，以竞争与传入的HPV结合，从而验证了一个全新的 预防病毒感染的方法。我们将开发第一个选择HPV突变体的遗传系统 信息丰富的表型，尝试识别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，并彻底改变了HPV的遗传分析。 至关重要的是，我们的研究将为基本细胞生物学提供新的见解。如果该提议是资助的，我们将 多年来，继续对病毒学和癌症研究做出新颖而重要的贡献。