Discovery and Characterization of Methylation Markers

描述(申请人提供)：肿瘤抑制基因(TSG)通过遗传或表观遗传事件的缺失或沉默已被发现在不同癌症的发展中发挥重要作用。早期研究表明，在膀胱尿路上皮细胞癌(UCC)中，TSG通过启动子高甲基化而频繁沉默是新的诊断和治疗靶点。在这里，我们建议进一步阐明膀胱癌的甲基化基因，从而确定新的甲基化基因作为早期检测、预后分类和预测膀胱尿路上皮细胞癌(UCC)治疗反应的标志。既然我们已经报道了甲基化基因在顺铂耐药发展中的作用，我们也将确定与顺铂反应相关的基因，并在UCC中被启动子高甲基化沉默。我们提出了三个具体目标：在具体目标1中，将采取综合筛选方法，在5个UCC细胞系中进一步鉴定因启动子高甲基化而沉默的新的肿瘤特异性基因。具体目的2我们将在有疾病和无疾病的患者的不同组织和体液中检测UCC的多个启动子超甲基化标志物，以建立简单的敏感性和特异性评估。最后，在特定的目标3中，我们将检查新基因在原发肿瘤中的功能意义和临床相关性，为未来的预防方法提供参考。将采用一种综合的方法，包括表达阵列杂交和药理揭示策略，以及Infinium甲基化试验，以识别膀胱癌进化中常见的表观遗传学改变。在体外和体内鉴定被启动子高甲基化失活的基因将有助于我们了解它们对UCC发生的影响，在肿瘤进展中的作用以及在耐药中的生物学作用。最终，已识别的膀胱癌特异性甲基化标志物将被用作非侵入性分子检测方法的标志物，PT1肿瘤的预后标志物，铂反应的测定，以及作为治疗预防的新靶点。

DESCRIPTION (provided by applicant): Deletion or silencing of tumor suppressor genes (TSG) through genetic or epigenetic events has been found to play an important role in the development of different cancers. Early studies suggest that frequent silencing of TSG through promoter hypermethylation in bladder Urothelial Cell Carcinomas (UCC) represent novel diagnostic and therapeutic targets. Here we propose to further elucidate the bladder cancer methylome and thereby identify novel methylated genes as markers for early detection, prognostic classification, and predictive classification of response to therapy of urothelial cell carcinoma of the bladder (UCC). Since we have reported a role for methylated genes in the development of cisplatin resistance, we will also identify genes that are related to cisplatin response and silenced by promoter hypermethylation in UCC. We are proposing three specific aims: In Specific Aim 1, an integrated screening approach will be undertaken to further identify novel tumor-specific genes silenced by promoter hypermethylation in 5 UCC cell lines. Specific Aim 2 We will test multiple promoter hypermethylation markers for UCC in various tissues and bodily fluids from patients with and without disease to establish simple sensitivity and specificity estimates. Finally, in Specific Aim 3 we will examine the functional significance and clinical relevance of the novel genes in primary tumors for future prevention approaches. A comprehensive approach consisting of expression array hybridization and pharmacological unmasking strategies, and the Infinium Methylation Assay will be taken to identify common epigenetic alterations in bladder tumor evolution. In vitro and in vivo characterization of identified genes which are inactivated by promoter hypermethylation will help us understand their impact on UCC development, role in cancer progression and biological role in drug resistance. Ultimately, identified bladder cancer specific methylation markers will be used as markers for non-invasive molecular detection approaches, prognostic markers for PT1 tumors, determination of platinum response and as novel targets for therapeutic prevention.