Characterization Of Wilms Tumor Suppressor Gene Product Wt1

Wilms 肿瘤抑制基因产物 Wt1 的表征

• 批准号: 8148846
• 负责人: Sean Lee
• 金额: $ 37.95万
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8148846
• 关键词: Nephroblastoma; Proteins; Tumor Suppressor Genes

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.

Wilms肿瘤抑制基因WT 1在10-15%的Wilms肿瘤（一种儿科肾癌）中发生突变，编码一种具有C2 H2锌指DNA结合结构域的转录因子。WT 1被提议调节对于肾、性腺、脾和肾上腺的启动和分化至关重要的基因的转录，因为WT 1缺失的小鼠胚胎缺乏所有这些器官。除了肾母细胞瘤，其他人类综合征和疾病也是由WT 1基因突变引起的，如Danys-Drash综合征和Frasier综合征。该项目的目标是：目标1。利用基因芯片表达谱分析技术筛选WT 1靶基因，并采用独立的方法对结果进行验证。目标2.将进一步测试经验证的靶标在kindey发育中的作用。为此，我们将通过RNA原位或免疫组织化学方法检测肾脏发育过程中靶基因的表达模式。此外，还将利用体外肾脏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和中和抗体的组合来实现。目标3. WT 1靶基因在肾母细胞瘤中的作用将进一步检查所确定的靶点在肿瘤发生中的作用。这将通过观察肿瘤样品中的表达水平和突变来完成。然后将在细胞系中进一步测试候选靶标的可能转化和生长促进特性。鉴定靶基因并确定其在发育过程中的作用将为肾母细胞瘤的发展和器官发生提供进一步的见解。