Clin.Relevance of Circulat.Tumor Cells in Bladder Cancer

描述(由申请人提供)：本提案的主要目的是确定检测膀胱癌患者循环肿瘤细胞(CTC)的临床相关性。其具体目的是：1)比较浅表性膀胱癌和转移性膀胱癌中尿路蛋白(I-11)、角蛋白(K19和K20)和表皮生长因子受体(EGFR)的CTC检出率；2)探讨这些分子标志物在原发和转移组织中的表达与CTC检出率的关系。我们建议使用逆转录聚合酶链式反应(RT-PCR)检测50例浅表肿瘤(预期微转移可能性较低)和50例严重转移性疾病(预期微转移可能性高)的膀胱癌患者的血液样本。所有患者将在四周内进行三次采样，以提高CTC阳性检测率。然后，CTC RT-PCR将与检测时是否存在疾病相关联，以确定对微转移的存在提供最佳敏感性、特异性和总体预测价值的最具临床信息量的标志物(S)。分子标志物的组织表达将使用免疫组织化学和RT-PCR对从AIM 1概述的患者中获得的样本进行评估，然后将结果与CTC的RT-PCR结果相关联。这一分析将有助于我们了解这些标记物在膀胱癌进展过程中组织表达的变化，以及这些变化与血液中CTC种植的关系。我们的初步数据清楚地表明，作为纽约大学医学院和纪念斯隆-凯特琳癌症中心合作开展这项工作的可行性。这种合作使这项研究拥有成功进行这类翻译研究所必需的强大的实验室和临床设施。我们计划将该项目作为第一阶段的试点研究，以响应“泌尿外科试点和可行性计划”计划公告#02-013；我们打算产生足够的数据，以便在第二阶段的前瞻性研究中继续研究CTC的发展，该研究将检验CTC检测与肌肉侵袭性疾病患者手术切除后复发之间的相关性。我们预计第三阶段标志物的开发将是一项多机构研究，目标是在考虑向FDA申请将CTC检测作为膀胱癌患者护理管理标准的一部分之前，独立验证和确认这种相关性。

DESCRIPTION (provided by applicant): The main objective of this proposal is to define the clinical relevance of detecting circulating tumor cells (CTC) in bladder cancer patients. The specific aims are: 1) To compare the detection rates of CTC for uroplakins (I-Ill), keratins (k19 and k20), and Epidermal Growth Factor Receptor (EGFR) in patients with superficial bladder cancer versus those with metastatic bladder cancer, and 2) To examine the relation between the expression of these molecular markers in primary and metastatic tissue deposits and their CTC detection rate. We propose to test blood samples from 50 bladder cancer patients with superficial tumors (expected to have low likelihood of micrometastases) and 50 with grossly metastatic disease (expected to have a high likelihood of micrometastases) using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). All patients will be sampled three times over a four week period to increase the positive CTC detection rate. The CTC RT-PCR will be then correlated with presence of disease at the time of the assay to define the most clinically informative marker(s) that provide the best sensitivity, specificity and overall predictive value for the presence of micrometastases. The tissue expression of the molecular markers will be assessed using immunohistochemistry as well as RT-PCR in specimens obtained from patients outlined in aim 1, and the results will then be correlated with the RT-PCR results for CTC. This analysis will help us to understand changes in the tissue expression of these markers during the progression of bladder cancer and how these changes are related to seeding of CTC in the blood. Our preliminary data clearly demonstrate the feasibility of conducting the proposed work as a collaborative effort between New York University School of Medicine and Memorial Sloan-Kettering Cancer Center. This cooperative work endows the study with the strong laboratory and clinical facilities that are necessary for the successful conduct of this type of translation research. We plan this project as a phase I pilot study, in response to the "Pilot and Feasibility Program in Urology" Program Announcement #02-013; we intend to generate sufficient data in order to pursue the development of CTC in a phase II prospective study that will examine the correlation between CTC detection and recurrence in patients with muscle invasive disease following surgical resection of the tumor. We foresee phase III marker development as a multi-institutional study with the goals of independently verifying and validating this correlation prior to considering an application to the FDA for integrating CTC detection as part of the standard of care management for bladder cancer patients.