P1 - Clinical Correlations of WTX Inactivation in Wilms Tumor

P1 - 肾母细胞瘤中 WTX 失活的临床相关性

• 批准号: 8517015
• 负责人: Daniel A. Haber
• 金额: $ 24.94万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2015-09-17
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8517015
• 关键词: 16q; Accounting; Affect; Age; Antibodies; Biological Markers; Biologically Based Therapy; Cell Line; Childhood; Childhood Renal Neoplasm; Clinical; Clinical Management; Complement; Dana-Farber Cancer Institute; Defect; Denys-Drash Syndrome; Development; Diagnosis; Diagnostic Neoplasm Staging; Disease; Disease Outcome; Embryo; Epigenetic Process; Frasier Syndromes; Future; Gene Mutation; Gene Silencing; Generations; Genes; Genetic; Genitourinary system; Goals; Histology; Immunoprecipitation; In Vitro; Insulin-Like Growth Factor II; Kidney; Kidney Neoplasms; Knockout Mice; Knowledge; Laboratories; Malignant Neoplasms; Mass Spectrum Analysis; Modeling; Molecular; Mus; Mutation; Nephroblastoma; North America; Outcome; Pathway interactions; Primary Neoplasm; Prognostic Factor; Prognostic Marker; Protein Binding; Proteins; Public Health Applications Research; Renal carcinoma; Resolution; Specimen; Stage at Diagnosis; Testing; Tissue Microarray; Treatment Protocols; Tumor Markers; Tumor Suppressor Genes; Tumor Suppressor Proteins; Tumor Tissue; Tumor stage; Validation; WT1 gene; Work; X Chromosome; authority; base; beta catenin; clinical application; clinically significant; comparative genomic hybridization; design; in vivo; in vivo Model; malformation; nephrogenesis; novel; outcome forecast; polyclonal antibody; prognostic; protein expression; success; tumor; tumorigenesis

Wilms tumor is the most common pediatric kidney cancer and is closely connected to kidney development. Mutations in two genes, WT1 and beta-catenin, and epigenetic changes in the insulin-like growth factor 2 (IGF2) locus have been described but the genetic basis of the majority of cases remains unknown. Given that current treatment protocols for Wilms tumor achieve high success rates (85%), there is a pressing need for prognostic markers that can guide clinical management but these have been difficult to define. We have recently identified a novel tumor suppressor located on the X chromosome, WTX, which is inactivated in 30% of Wilms tumor cases. We now propose to build on our initial findings to establish clinical correlates of WTX inactivation and to test the potential application of WTX mutations as markers of prognosis. We will also define additional markers by identifying molecular pathways that are affected by WTX inactivation Together, these studies will achieve immediate translational goals by defining novel biomarkers in Wilms tumor and will further our understanding of this disease to allow the future development of biologically based therapies. Specific aims: 1) To define clinical correlations of WTX inactivation in Wilms tumor. We will analyze 200 Wilms tumors for WTX mutations and correlate our findings with disease outcomes and other clinical parameters such as age at presentation, stage at diagnosis, bilaterality and associated developmental malformations. We will also develop a WTX polyclonal antibody and a Wilms tumor tissue microarray to test WTX protein levels. 2) Modeling WTX function to identify pathways of potential clinical significance. We will use immunoprecipitation of tagged WTX followed by mass spectrometry to define interactions between WTX and other cellular components. The functional consequences of these interactions will be validated in vitro using kidney derived cell lines and in vivo using a WTX conditional knockout mouse. 3) Clinical validation of novel Wilms tumor markers. Genes involved in WTX related pathways will be tested for potential clinical applications as biomarkers by correlating expression levels with clinical parameters. We will also test selected genes for mutations in primary Wilms tumors. We anticipated that, by defining prognostic markers and furthering our knowledge of WTX related pathways in Wilms tumor, this project will have public health applications in the treatment of pediatric kidney cancer.

威尔姆斯肿瘤是最常见的小儿肾癌，与肾脏发育密切相关。已经描述了两个基因WT1和β-catenin的突变，以及胰岛素样生长因子2（IGF2）基因座的表观遗传变化，但大多数病例的遗传基础仍然未知。鉴于Wilms肿瘤的当前治疗方案达到了很高的成功率（85％），因此对预后标记的迫切需求可以指导临床管理，但这些标记很难定义。我们最近确定了一个位于X染色体WTX上的新型肿瘤抑制剂，该肿瘤抑制剂在30％的Wilms肿瘤病例中被灭活。现在，我们建议建立我们的初步发现，以建立WTX失活的临床相关性，并测试WTX突变作为预后标志的潜在应用。我们还将通过确定受WTX失活影响的分子途径来定义其他标记，这些研究将通过定义Wilms肿瘤的新生物标志物来实现立即的转化目标，并将进一步了解我们对这种疾病的理解，从而允许未来基于生物学的疗法发展。具体目的：1）定义WILMS肿瘤中WTX失活的临床相关性。我们将分析200个WILMS肿瘤，以进行WTX突变，并将我们的发现与疾病结果和其他临床参数（例如年龄，诊断阶段，双侧性及相关发育畸形）相关。我们还将开发WTX多克隆抗体和Wilms肿瘤组织微阵列以测试WTX蛋白水平。 2）建模WTX功能以识别潜在临床意义的途径。我们将使用标记的WTX的免疫沉淀，然后使用质谱法来定义WTX与其他细胞组件之间的相互作用。这些相互作用的功能后果将在体外使用肾脏衍生的细胞系和在体内使用WTX条件基因敲除小鼠进行验证。 3）新型Wilms肿瘤标记的临床验证。通过将表达水平与临床参数相关联，将测试参与WTX相关途径的基因作为生物标志物的潜在临床应用。我们还将测试选定的基因在原发性威尔姆斯肿瘤中的突变。我们预计，通过定义预后标记并进一步推动WILMS肿瘤中与WTX相关途径的了解，该项目将在治疗儿科肾脏癌症治疗时采用公共卫生应用。