Virus-Host Interactions that Modulate Early Steps of Human Papillomavirus Infecti

描述(由申请人提供)：我们研究的长期目标是了解在HPV相关的癌症进展过程中建立和维持人类角质形成细胞中HPV感染所必需的HPV-宿主相互作用的机制。人乳头瘤病毒(HPV)是最常见的性传播病原体，与越来越多的人类癌症有关，包括宫颈癌、阴道癌、阴茎癌、肛门癌、头颈部癌和其他鳞癌，每年导致全球近50万人死亡(国际癌症研究机构)。要建立持续感染，病毒必须进入细胞核，并将其基因组建立为核质粒。然而，由于技术限制，人们对HPV基因组如何进入细胞核并建立其复制知之甚少。为了实现我们的目标，我们将：1)确定宿主细胞有丝分裂和微管重组在人类角质形成细胞HPV DNA核进入中的作用；2)研究宿主的先天反应及其在体外和体内原代角质形成细胞HPV感染早期步骤中的作用。在这些研究中，我们将利用感染性HPV病毒粒子和实时成像技术，开发具有各种荧光标记的可靠HPV DNA标记系统，并跟踪HPV DNA在活体人角质形成细胞中的运动。此外，我们还将通过敲除参与有丝分裂、微管网络和宿主先天免疫的宿主因子来测试HPV感染性、DNA进入细胞核和基因组复制的变化。最近开发的预防性HPV疫苗最终应该会减少某些HPV基因型的感染。然而，由于疫苗不能覆盖所有的HPV，也不能消除现有的感染，仍然迫切需要开发干预HPV感染和防止癌症进展的新方法。我们的研究将进一步了解HPV用于建立感染的机制，从而确定靶向预防HPV相关癌症的途径。

DESCRIPTION (provided by applicant): The long-term goals of our research are to understand the mechanisms of HPV-host interactions necessary for establishing and maintaining HPV infections in human keratinocytes throughout HPV-associated cancer progression. Human papillomaviruses (HPVs) are the most common sexually transmitted pathogen, causally associated with a growing number of human cancers including cervical, vaginal, penile, anal, head/neck, and other squamous carcinomas, causing almost half a million deaths worldwide each year (International Agency for Research on Cancer). To establish persistent infections, the virus must enter the nucleus and establish its genome as a nuclear plasmid. However, little is known about how HPV genome enters the nucleus and establishes its replication, due to technical limitations. To achieve our goals, we will: 1) define the role of host cell mitosis and microtubule reorganization for HPV DNA nuclear entry in human keratinocytes; and 2) investigate host innate responses and its roles for early steps of HPV infection in primary keratinocytes in vitro and in vivo. In these studies, we will develop reliable HPV DNA labeling systems with various fluorescent tags and track the movement of HPV DNA in live human keratinocytes, using infectious HPV virions and real time imaging technology. In addition, we will test alterations of HPV infectivity, DNA entry into the nucleus, and genome replication by knockdown of host factors involved in mitosis, microtubule network, and host innate immunity. Recently developed prophylactic HPV vaccines should eventually reduce infections by some HPV genotypes. However, because the vaccines do not cover all HPVs and do not eliminate existing infections, there remains an urgent need to develop new means of intervening in HPV infections and preventing cancer progression. Our studies will provide further understanding of mechanisms employed by HPVs to establish infections and thereby identify pathways to target for preventing HPV-associated cancers.