TUMOR SUPPRESOR GENE PROMOTER HYPERMETHYLATION FOR DETECTION OF HEAD AND NECK

我们和其他人已经证明了肿瘤抑制基因的沉默与启动子相关 高甲基化是许多人类癌症的共同特征。我们发现了几种新的肿瘤 启动子超甲基化失活的抑制基因以及用于检测的候选基因 战略。我们建议继续鉴定和表征鳞状细胞中的高甲基化基因。 并将通过以下方式鉴定新的启动子高甲基化基因：(1)A 基于检测在其他肿瘤中被发现高甲基化的基因的候选基因方法 候选基因的功能结构和生物学合理性；(2)功能筛选 用脱甲基剂(5-氮杂脱氧胞苷)处理后上调的基因。新开 已确定的高甲基化基因将被表征为功能意义和生物活性 肿瘤进展。我们将应用一种改进的药理去掩蔽方法来定义启动子 高甲基化基因的失活导致对传统化疗药物的耐药性 和途径(EFGR)导向治疗。候选的高甲基化肿瘤抑制基因将是 通过在体外和体内小鼠模型中导入候选基因，在耐药细胞系中得到验证。 然后，这些基因将在前瞻性试验和回溯性队列中得到验证，这些队列使用传统的 化疗和EGFR途径特异性药物。新的启动子超甲基化靶点将是 纳入第二个孢子提案(项目2)，用作早期发现和 对头颈部癌症患者的监测。

We and others have demonstrated that silencing of tumor suppressor genes associated with promoter hypermethylation is a common feature of many human cancers. We have identified several novel tumor suppressor genes inactivated by promoter hypermethylation, as well as candidate genes for use in detection strategies. We propose continue to identify and characterize hypermethylated genes in squamous cell carcinoma of the head neck (HNSCC) and will identify novel promoter hypermethylated genes by: (1) A candidate gene approach based on testing genes found to be hypermethylated in other tumors or on the functional structure and biologic plausibility of the candidate gene and (2) A functional screen looking for genes that are up-regulated following treatment with demethylating agents (5-Aza-deoxycytidine). Newly identified hypermethylated genes will be characterized for functional significance and biologic activity in tumor progression. We will apply a modified pharmacologic unmasking approach to define promoter hypermethylated genes whose inactivation contributes to drug resistance to conventional chemotherapeutic and pathway (EFGR) directed therapy. Candidate hypermethylated tumor suppressor genes will be validated in drug resistant cell lines by transfection of candidate genes in in vitro and in vivo mouse models. These genes will then be validated in prospective trials and retrospective cohorts treated with conventional chemotherapy and EGFR pathway specific agents. Novel promoter hypermethylatedtargets will be incorporated into a second SPORE proposal (project #2) for use as markers for the early detection and monitoring of patients with head and neck cancer.