Factors Influencing KSHV Infection and Spread within the Oral Cavity

 描述（由申请方提供）：卡波西肉瘤疱疹病毒（KSHV）是一种致癌性人疱疹病毒，在缺乏充分免疫控制的情况下可引起卡波西肉瘤（KS）以及B细胞淋巴瘤。KSHV相关肿瘤是移植患者和HIV患者发病率和死亡率的重要原因。述口腔是 KSHV复制和持续存在的重要位点，唾液被认为是人与人之间传播的关键来源。此外，KS的口腔表现与快速进展和高死亡率相关。KSHV相关肿瘤是多灶性的，并且经常是多克隆的，这表明持续的病毒感染对疾病进展很重要。由于KSHV分子病毒学特有的技术挑战，对生理相关细胞类型的感染知之甚少。目前的项目将利用新的分子遗传学工具来探索KSHV感染的基本病毒学，目的是确定新的病毒和细胞靶点，以防止口腔内的病毒传播和KS进展。在K99阶段，我们将建立KSHV易感细胞类型和组织的从头感染系统，以探索感染、持久性和传播的细胞类型特异性。然后，我们将利用这些模型来探索病毒FLICE抑制蛋白（vFLIP）在KSHV感染背景下的作用。为了将这些研究放在最相关的疾病背景下，我们将酌情包括艾滋病毒合并感染，以确定这两种病原体之间的基本相互作用。随着关键培训和职业发展的完成，模型系统在手，vFLIP研究作为在这些模型中使用重组KSHV毒株的原理证明，我将在R00阶段向前推进，以检查病毒体相关的非结构KSHV蛋白对相关细胞类型感染的最早阶段的贡献。这些打包的、非结构性的 蛋白质包含称为被膜的病毒体结构。被膜因子在病毒进入后立即影响宿主细胞生理学的深刻改变，并且在其他疱疹病毒系统中对这些因子的研究已经产生了对病毒和细胞生物学的重要见解。然而，KSHV被膜的组成和功能仍然不清楚，代表了我们对KSHV病毒学基础的理解存在很大差距。这项研究的结果将提供有关口腔中KSHV持续存在的动态过程的关键信息，并提供对病毒和细胞机制的见解，可用于新的治疗模式。使用广泛而系统的工作流程研究多个KSHV皮层因素，我将获得一个重要的机构，既可验证的数据和初步数据，为多个未来的研究，这将提供一个坚实的基础上，建立一个独立的 研究生涯。

 DESCRIPTION (provided by applicant): Kaposi sarcoma herpesvirus (KSHV) is an oncogenic human herpesvirus, which causes Kaposi sarcoma (KS) as well as B cell lymphomas in the absence of adequate immune control. KSHV-associated tumors are a significant cause of morbidity and mortality in transplant patients and individuals with HIV-disease. The oral cavity is an important site for KSHV replication and persistence and saliva is believed to be a critical source of person-to-person transmission. Moreover, oral manifestation of KS is associated with rapid progression and high mortality rates. KSHV-associated tumors are multi-focal and frequently polyclonal, suggesting that continued viral infection is important for disease progression. Because of technical challenges unique to KSHV molecular virology, little is known about infection of physiologically relevant cell types. The current project will leverage new molecular genetics tools to explore the fundamental virology of KSHV infection with the goal of identifying novel viral and cellular targets to prevent viral spread and KS progression within the oral cavity. During the K99 phase, we will establish systems for de novo infection of KSHV-susceptible cell types and tissues in order to explore the cell type-specific properties of infectin, persistence and spread. We will then utilize these models to explore the contribution of the viral FLICE- inhibitory protein (vFLIP) in the context of KSHV infection. In order to place these studies in the most disease-relevant context possible, we will include HIV co-infection, as appropriate, to determine fundamental interactions between these two pathogens. With critical training and career development accomplished, model systems in hand, and the vFLIP studies as proof-of-principle for the use of recombinant KSHV strains within these models, I will move forward in the R00 phase to examine the contribution of virion-associated non-structural KSHV proteins to the earliest stages of infection in relevant cell types. These packaged, non-structural proteins comprise a virion structure known as the tegument. Tegument factors affect profound alterations in host cell physiology immediately upon virus entry and studies of these factors in other herpesvirus systems have yielded important insights into viral and cellular biology. However, the composition and function of the KSHV tegument remains obscure and represents a large gap in our understanding of fundamental KSHV virology. The results of this research will provide critical information about the dynamic process of KSHV persistence in the oral cavity and provide insights into both viral and cellular mechanisms that can be exploited for new treatment paradigms. Using a broad but systematic workflow of study for multiple KSHV tegument factors, I will obtain a significant body of both publishable data and preliminary data for multiple future studies that will provide a solid foundation upon which to build an independent research career.