MOLECULAR GENETICS OF WILMS' TUMOR

这一构成部分的基本目标将继续利用遗传 寻找和表征在生物多样性研究中具有重要意义的基因的策略 人类肿瘤的发展以及肿瘤对 治疗。该计划的主要重点一直是，并将继续是 肾母细胞瘤(肾母细胞瘤)的发生过程。我们会 继续研究肾母细胞瘤作用的分子基础 抑制基因WT1是在过去的一段时间里发现并鉴定的 授权期。我们将扩大我们的研究，涉及基因打靶 为了解WT1基因在小鼠ES细胞中的作用 肿瘤的发生和正常发育以及鉴定功能 WT1多肽的选择性剪接形式的作用。我们会 继续WT1的生化和功能特征 扩展我们在WT1多肽相互作用伙伴方面的工作 酵母双杂交系统，以表征WT1与其他 多肽利用免疫化学，开发基于细胞的系统以 直接证明WT1对特定靶基因的影响 利用体外转录和剪接系统进行替代 WT1多肽的形式。为了更深入地了解WT1的功能， 我们将探索WT1在肾脏以外的系统中的作用，如 造血术。我们将完成对 WT2抑癌基因的定位克隆策略 人类肿瘤的突变和缺失分析。我们将进一步 分析细胞凋亡在肿瘤治疗反应中的作用 基于遗传学的方法，如对肿瘤细胞的进一步研究 缺乏P53。这些研究策略将提供框架 用于基因鉴定和表征的范围超出了 直接省的Wilms肿瘤与其他共有的肿瘤类型 肾母细胞瘤的致瘤机制及其与临床的关系 研究化疗药物反应的潜在基础 以及使用在此期间开发的遗传方法的扩展的辐射 上一个授权期。

The basic objectives of this component will continue to utilize genetic strategies to identify and characterize genes of major significance in the development of human tumors as well as the response of tumors to treatment. The main focus of this program has been and continues to be the process of tumorigenesis in nephroblastoma (Wilms tumor). We will continue to study the molecular basis of the action of the Wilms tumor suppressor gene, WT1, discovered and characterized during the previous granting period. We will extend our studies involving gene targeting of mouse ES cells in order to understand the role of the WT1 gene in tumorigenesis and normal development as well as to identify the functional roles of alternatively spliced forms of the WT1 polypeptide. We will continue the biochemical and functional characterization of WT1 by extending our work on interaction partners for the WT1 polypeptide using the yeast two hybrid system, to characterize WT1 interactions with other polypeptides using immunochemistry, to develop cell based systems to directly demonstrate the effects of WT1 on specific target genes and to utilize in vitro transcription and splicing systems for the alternate forms of the WT1 polypeptide. To gain deeper insight into WT1 function, we will explore the role of WT1 in systems other than kidney such as hematopoiesis. We will complete identification and characterization of the WT2 tumor suppressor gene using positional cloning strategies and analysis of mutations and deletions in human tumors. We will further analyze the role of apoptosis in tumor treatment response using genetically based approaches such as further study of the tumor cells deficient for p53. These research strategies will provide the framework for gene identification and characterization extending beyond the immediate province of Wilms tumor to other tumor types which share common mechanisms of tumorigenesis with Wilms tumor as well as to further investigate the underlying basis of response to chemotherapeutic agents and radiation using extensions of the genetic approaches developed during the previous grant period.