Discovery and Characterization of Methylation Markers

甲基化标记的发现和表征

• 批准号: 8495291
• 负责人: DAVID SIDRANSKY
• 金额: $ 31.6万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8495291
• 关键词: Affect; Biological; Biological Assay; Biological Markers; Biology; Bladder Neoplasm; Bladder Urothelial Cell; Bladder Urothelium; Candidate Disease Gene; Carcinogens; Carcinoma; Cell Line; Characteristics; Cisplatin; Classification; Clinical; DNA; DNA Methylation; Data; Detection; Development; Disease; Drug resistance; Early Diagnosis; Environmental Exposure; Epigenetic Process; Event; Evolution; Exposure to; Fingerprint; Future; Gene Silencing; Gene Targeting; Genes; Genetic; Histone Deacetylase Inhibitor; Hybridization Array; Hypermethylation; In Vitro; Incidence; Individual; Lead; Lesion; Liquid substance; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Methylation; Molecular; Monitor; Neoplastic Cell Transformation; Passive Smoking; Pathogenesis; Pathway interactions; Patients; Performance; Pharmaceutical Preparations; Phase; Platinum; Play; Prevention; Prevention approach; Primary Neoplasm; Process; Prognostic Marker; RNA; Recurrence; Reporting; Resistance; Role; Sampling; Sensitivity and Specificity; Sequence Analysis; Staging; Stratification; Techniques; Testing; Therapeutic; Time; Tissues; Tumor Suppressor Genes; Urothelial Cell; Validation; assay development; base; bladder Carcinoma; case control; clinical phenotype; clinically relevant; in vivo; methylome; neoplastic; new therapeutic target; novel; novel diagnostics; outcome forecast; prognostic; promoter; response; screening; translational approach; tumor; tumor progression

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.

描述（由申请人提供）：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起重要作用。早期研究表明，膀胱尿路上皮细胞癌（UCC）中TSG通过启动子超甲基化而频繁沉默代表了新的诊断和治疗靶点。在此，我们建议进一步阐明膀胱癌甲基组，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）治疗反应分类的标记。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，我们也将确定与顺铂反应相关的基因，并在UCC中被启动子超甲基化沉默。我们提出了三个具体目标：在specific Aim 1中，将采用综合筛选方法进一步鉴定5种UCC细胞系中被启动子超甲基化沉默的新型肿瘤特异性基因。我们将在患有和非疾病患者的各种组织和体液中检测UCC的多个启动子超甲基化标记，以建立简单的敏感性和特异性估计。最后，在Specific Aim 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，为未来的预防方法提供帮助。一个综合的方法包括表达阵列杂交和药理学揭示策略，以及Infinium甲基化分析，以确定膀胱肿瘤进化中常见的表观遗传改变。在体外和体内鉴定被启动子超甲基化灭活的基因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中的生物学作用。最终，确定的膀胱癌特异性甲基化标记物将用作无创分子检测方法的标记物、PT1肿瘤的预后标记物、铂反应的测定以及治疗预防的新靶点。