Project 1: Role of HTLV-1 Hbz in Transformation and Disease

项目摘要--项目1 对逆转录病毒的研究导致了重要的发现，并导致了对基本细胞生物学的洞察，包括 细胞信号的机制，基因表达的调节，最终细胞转化和癌症。 我们在PPG内的合作重点是HTLV-1，它主要与成人T细胞有关 白血病(ATL)和神经系统疾病(HAM/TSP)在一小部分感染者中。疾病 HTLV-1的进展被归因于税收，尽管我们和其他人已经假设并提供了数据 另一种被称为Hbz的病毒基因在恶性过程中发挥着关键作用。我们提供的数据也是如此 由于其他人的工作，表明hbz是多功能的，在rna和蛋白质中都发挥着重要的作用。 在整个感染和潜伏期的建立过程中形成。我们已经通过PPG扩展了我们提议的工作 合作以更好地了解Hbz促进细胞增殖和生存活动的方式。 蛋白质组学分析确定了一组与hbz RNA和hBZ蛋白明显相互作用的细胞蛋白质。 这些相互作用的功能表征是下一个关键步骤，也是高度集成的基础 旨在进一步确定红血球在转化和疾病中的作用的延续项目。我们的整体 假设发现hbz rna和hbz蛋白的作用机制将提供重要的 对HTLV-1细胞转化和疾病的洞察，最终将为治疗提供手段 旨在根除HTLV-1在宿主中的持久性。此项目引用的高度集成的建议书 下面的核心协作有两个具体目标。目的1剖析hbz RNA的作用机制(S)。 并确定其在致病过程中的作用。我们假设细胞蛋白与 Hbz mRNA20结构转译为细胞信号通路，对病毒的持久性和细胞 扩散。我们新的蛋白质组学数据已经确定并进一步验证了结合不同细胞蛋白的 Hbz基因。这一目标将利用体外方法(项目2和3，核心管理/生物统计和病毒载体) 鉴定hbz RNA相互作用区并确定细胞结合蛋白在细胞中的功能作用 扩散。我们将使用体内方法(动物核心)来了解RNA和 相互作用蛋白在持久性和肿瘤形成中的建立。《目标2》将剖析其中的机制(S) 研究HBZ蛋白的作用并确定其在HTLV-1病理生物学中的作用。我们新的蛋白质组学数据 鉴定并进一步验证了与HBZ结合的不同细胞蛋白。体外方法(项目3，核心 Admin/Biostats和病毒载体)，包括蛋白质相互作用图谱、细胞增殖分析和 将利用细胞转化分析来确定HBZ互动区和KEY的功能作用 HTLV-1病理生物学中的细胞结合蛋白。活体方法(动物核心，项目2)将用于 确定这些HBZ相互作用蛋白在建立持续性感染细胞中的作用 生存和肿瘤形成。

PROJECT SUMMARY – PROJECT 1 The study of retroviruses has resulted in important discoveries and led to insights into basic cell biology including mechanisms of cell signaling, regulation of gene expression, and ultimately cellular transformation and cancer. Our collaborative work within this PPG focuses on HTLV-1, which is associated primarily with adult T-cell leukemia (ATL) and neurological disease (HAM/TSP) in a small percentage of infected individuals. Disease progression by HTLV-1 has been attributed to Tax, although we and others have hypothesized and provide data that another viral gene, termed Hbz, plays a critical role in the malignant process. Our presented data, as well as the work of others, indicate that hbz is multi-functional, playing important roles in both the RNA and protein form throughout infection and establishment of latency. We have extended our proposed work through PPG collaborations to better understand the manner in which hbz promotes proliferative and survival activity in cells. Proteomic analyses identified a panel of cellular proteins that distinctly interact with hbz RNA and HBZ protein. Functional characterization of these interactions are the next critical steps and the basis for this highly integrative continuation project designed to further define the role of hbz in transformation and disease. Our overall hypothesis is that uncovering the mechanism of actions of hbz RNA and HBZ protein will provide important insight into HTLV-1 cellular transformation and disease and ultimately will provide means for therapeutic targeting to eradicate HTLV-1 persistence in the host. This highly integrated proposal as referenced by Project and Core collaborations below has two Specific Aims. Aim 1 will dissect the mechanism(s) of action of hbz RNA and determine its contributions to the pathogenic process. We hypothesize that cellular protein interaction with hbz mRNA 2o structure translates to cell signaling pathways important for viral persistence and cellular proliferation. Our new proteomics data has identified, and we further validated, distinct cellular proteins that bind hbz mRNA. This aim will utilize in vitro approaches (Projects 2 and 3, Cores Admin/Biostats and Viral Vector) to identify the hbz RNA interactive region and determine the functional role of cellular binding proteins in cell proliferation. We will use in vivo approaches (Animal Core) to understand the contribution of the RNA and interactive proteins in the establishment of persistence and tumor formation. Aim 2 will dissect the mechanism(s) of action of HBZ protein and determine its contributions to HTLV-1 pathobiology. Our new proteomics data has identified, and we further validated, distinct cellular proteins that bind HBZ. In vitro approaches (Project 3, Cores Admin/Biostats and Viral Vector) including protein interaction mapping, cellular proliferation assays, and cellular transformation assays will be utilized to identify the HBZ interactive region and the functional role of key cellular binding proteins in HTLV-1 pathobiology. In vivo approaches (Animal Core, Project 2) will be used to determine the contribution of these HBZ interactive proteins in the establishment of persistence, infected cell survival, and tumor formation.