Discovery and Characterization of Methylation Markers

描述（由申请人提供）：已发现通过遗传或表观遗传事件删除或沉默肿瘤抑制基因（TSG）在不同癌症的发生中发挥重要作用。早期研究表明，在膀胱尿路上皮细胞癌（UCC）中，通过启动子高甲基化频繁沉默TSG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甲基化组，从而确定新的甲基化基因作为标记物的早期检测，预后分类，并预测分类的膀胱尿路上皮细胞癌（UCC）的治疗反应。由于我们已经报道了甲基化基因在顺铂耐药发展中的作用，我们还将鉴定与顺铂反应相关的基因，并在UCC中通过启动子超甲基化沉默。我们提出了三个具体目标：在具体目标1中，将采用综合筛选方法进一步鉴定5种UCC细胞系中因启动子高甲基化而沉默的新型肿瘤特异性基因。具体目标2我们将测试多种UCC启动子超甲基化标志物在各种组织和体液从患者和非疾病，以建立简单的敏感性和特异性估计。最后，在具体目标3中，我们将研究新基因在原发性肿瘤中的功能意义和临床相关性，以用于未来的预防方法。 将采用由表达阵列杂交和药理学解蔽策略以及Infinium甲基化测定组成的综合方法来鉴定膀胱肿瘤演变中常见的表观遗传学改变。通过启动子超甲基化失活的已鉴定基因的体外和体内表征将有助于我们了解它们对UCC发展的影响、在癌症进展中的作用以及在耐药性中的生物学作用。最终，确定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.