Characterization Of Wilms Tumor Suppressor Gene Product Wt1

Wilms肿瘤抑制基因WT1在10-15%的Wilms肿瘤（一种儿童肾癌）中发生突变，编码一种具有C2H2锌指DNA结合域的转录因子。由于WT1缺失的小鼠胚胎缺乏所有这些器官，因此WT1被认为可以调节对肾脏、性腺、脾脏和肾上腺的启动和分化至关重要的基因的转录。除了Wilms肿瘤外，其他人类综合征和疾病也由WT1基因突变引起，如Danys-Drash综合征和Frasier综合征。这个项目的目标是：目标1。通过微阵列表达谱分析鉴定WT1靶基因，并使用独立的方法验证结果。目标2。经过验证的目标将进一步检验其在肾脏发展中的作用。为此，我们将通过RNA原位或免疫组织化学检测肾脏发育过程中靶基因的表达模式。此外，将利用体外肾器官培养系统来检测靶基因的功能作用。这将通过使用siRNA或shRNA的组合以及中和抗体来完成。目标3。WT1靶基因在Wilms肿瘤中的作用。已确定的靶点将进一步研究其在肿瘤发生中的作用。这将通过观察肿瘤样本中的表达水平和突变来完成。候选靶点将在细胞系中进一步测试可能的转化和促进生长的特性。鉴定靶基因并确定其在发育过程中的作用将为Wilms肿瘤的发展和器官发生提供进一步的见解。

The Wilms tumor suppressor gene WT1, which is mutated in 10-15% of Wilms tumor -- a pediatric kidney cancer -- encodes a transcription factor with C2H2 zinc finger DNA binding domain. WT1 is proposed to regulate transcription of genes that are critical for the initiation and differentiation of kidney, gonad, spleen, and adrenal gland since wt1-null mouse embryos lack all of these organs. In addition to Wilms tumor, other human syndromes and diseases are also caused by mutations in the WT1 gene such as Danys-Drash and Frasier syndromes. The Aims of this project are: Aim 1. To identify WT1 target genes by using microarray expression profiling analysis, and to validate the result using independent methods. Aim 2. Validated targets will be further tested for their role in kindey development. To this end, we will examine the expression patterns of the target genes by RNA in-situ or immunohistochemistry during kidney development. In addition, in vitro kidney organ culture system will be utilized to examine the functional role of the target genes. This will be accomplished by using the combination of siRNA or shRNA, and neutralizing antibodies. Aim 3. The roles of WT1 target genes in Wilms tumor. The identified targets will be further examined for their role in tumorigenesis. This will be done by looking at expression levels and mutations in the tumor samples. Candidate targets will then be further tested in cell lines for possible transformation and growth-promoting properties. Identification of the target genes and defining their role during development will provide further insights to the development of Wilms tumor and organogenesis in general.