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

项目摘要-项目1 逆转录病毒的研究已经导致了重要的发现，并导致了对基础细胞生物学的深入了解，包括 细胞信号传导机制、基因表达调控，最终导致细胞转化和癌症。 我们在这个PPG中的合作工作集中在HTLV-1，它主要与成人T细胞相关。 白血病（ATL）和神经系统疾病（HAM/TSP）在一小部分受感染的个体。疾病 HTLV-1的进展归因于税收，尽管我们和其他人已经假设并提供了数据 另一种病毒基因，Hbz，在恶性过程中起着关键作用。我们提供的数据，以及 表明hbz具有多功能，在RNA和蛋白质中均发挥重要作用 形成贯穿感染和潜伏期的建立。我们已经通过PPG扩展了我们的拟议工作 合作，以更好地了解HBZ促进细胞增殖和存活活性的方式。 蛋白质组学分析鉴定了一组与hbz RNA和HBZ蛋白明显相互作用的细胞蛋白。 这些相互作用的功能表征是下一个关键步骤，也是这种高度整合的基础。 旨在进一步确定hbz在转化和疾病中的作用的延续项目。我们的整体 假设揭示hbz RNA和HBZ蛋白的作用机制将提供重要的 深入了解HTLV-1细胞转化和疾病，并最终将提供治疗手段， 靶向根除宿主中的HTLV-1持久性。该项目引用的高度集成的提案 下面的核心合作有两个具体目标。目的1将剖析hbz RNA的作用机制 并确定其在致病过程中的作用。我们假设细胞蛋白质与 hbz mRNA 2 o结构翻译为对病毒持久性和细胞增殖重要的细胞信号传导途径 增殖我们的新蛋白质组学数据已经确定，我们进一步验证，不同的细胞蛋白质结合 hbz mRNA。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） 为了鉴定hbz RNA相互作用区域并确定细胞结合蛋白在细胞中的功能作用， 增殖我们将使用体内方法（动物核心）来了解RNA的贡献， 相互作用的蛋白质在建立持久性和肿瘤形成。目标2将剖析机制 HBZ蛋白的作用，并确定其对HTLV-1病理生物学的贡献。我们新的蛋白质组学数据 我们鉴定并进一步验证了结合HBZ的不同细胞蛋白。体外方法（项目3，核心 给药/生物统计和病毒载体），包括蛋白质相互作用图谱、细胞增殖测定和 细胞转化试验将用于鉴定HBZ相互作用区和关键的功能作用。 细胞结合蛋白在HTLV-1病理生物学中的作用。体内方法（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久性感染细胞中的作用， 存活和肿瘤形成。

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.