MOLECULAR GENETICS OF WILMS' TUMOR

这一部分的基本目标将继续利用遗传资源， 战略，以确定和表征基因的重大意义， 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（Wilms瘤）的肿瘤发生过程。 我们将 继续研究肾母细胞瘤作用的分子基础 抑制基因WT1，在以前的研究中发现并鉴定 授予期。 我们将扩大我们的研究，包括基因靶向， 小鼠ES细胞，以了解WT1基因在 肿瘤发生和正常发育，以及确定功能 WT1多肽的可变剪接形式的作用。 我们将 继续WT1的生物化学和功能表征， 扩展了我们对WT1多肽相互作用伴侣的研究， 酵母双杂交系统，以表征WT1与其他 多肽，开发基于细胞的系统， 直接证明WT1对特定靶基因的作用， 利用体外转录和剪接系统， 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， 造血 我们将完成对 WT2肿瘤抑制基因的定位克隆策略， 分析人类肿瘤中的突变和缺失。 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，如进一步研究肿瘤细胞， 缺乏p53。 这些研究战略将提供框架， 用于基因鉴定和表征， 威尔姆斯瘤与其他有共同点的肿瘤类型的直接关系 肾母细胞瘤的肿瘤发生机制，以及进一步 研究对化疗药物反应的潜在基础 和辐射使用的扩展的遗传方法开发期间 上一个补助期。

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.