A New Approach for the Non-Invasive Detection of Bladder Cancer

描述（申请人提供）：目前迫切需要开发一种有效的检测原发性和复发性膀胱癌的方法。膀胱癌是西方国家男性第四大常见恶性肿瘤，女性第九大常见恶性肿瘤。如果发现得早，膀胱癌的特点是生存率高（94%）。然而，如果已经扩散到其他器官，存活率会显著下降，低至14%。膀胱癌的另一个特点是复发率高（50% - 90%）。因此，膀胱癌患者必须经常接受膀胱镜检查。该项目的目标是探索一种无创尿液诊断测试的可行性，这种测试可以早期发现新发和复发的膀胱癌，并为癌症分期提供有效的工具。这样的测试，如果适用于高危人群或被调查的患者，可以显著提高生存率，并有助于改善生活质量和持续时间。包括膀胱镜检查和细胞学检查的常规诊断方法要么是侵入性的（膀胱镜检查），要么是相对不敏感的（细胞学检查）。我们建议应用一种基于体外表达蛋白质谱（MASSIVE- PRO）的新方法，该方法将能够检测膀胱癌患者尿液中存在的一小部分突变DNA。AmberGen已经进行了初步实验，证明了该提案在几个关键领域的可行性。在I期，将首先使用细胞系DNA样本的混合物，然后使用膀胱癌患者的临床样本，对MASSIVE-PRO系统进行相关基因突变检测的评估。目标是检测Kras/Hras、FGFR3（低分期/低分级肿瘤的标志物）和p53（高分期/高分级肿瘤的标志物）在样本中5%或更低水平发生的突变。一小组匹配的组织和尿液将由布里格姆妇女医院（Boston， MA）外科教授和泌尿学研究主任Kevin Loughlin医生提供。在II期，基于I期结果的分子检测将在临床实验室环境中进一步开发和评估，以检测早期膀胱癌。

DESCRIPTION (provided by applicant): There is an urgent need to develop an effective method for detection of primary and recurrent bladder cancer. Bladder cancer is the fourth most common malignancy in men and the ninth most common malignancy in women in Western countries. If identified early, bladder cancer is characterized by a high survival rate (94%). However, the survival rate drops significantly, to as low as 14%, if spread to other organs has occurred. Bladder cancer is also characterized by a high recurrence rate, (50% - 90%). Consequently, bladder cancer patients must undergo frequent cystoscopic surveillance. The goal of this project is to explore the feasibility of a noninvasive diagnostic test from urine that would allow early detection of new onset, as well as recurrent, bladder cancers, and which would provide an effective tool for cancer staging. Such a test, if applied to high-risk populations or surveyed patients could significantly increase survival rates, and could contribute to improved life quality and duration. Conventional diagnostic methods that involve cystoscopy and cytology are either invasive (cystoscopy), or are relatively insensitive (cytology). We propose to apply a new method based on mass spectrometry of in vitro expressed proteins (MASSIVE- PRO) that will enable detection of small fractions of mutated DNA present in the urine of bladder cancer patients. AmberGen has performed preliminary experiments that demonstrate feasibility in several key areas of this proposal. In Phase I, the MASSIVE-PRO system will be evaluated for detection of mutations in a panel of relevant genes first using mixtures of cell line DNA samples, and later using clinical samples from bladder cancer patients. The goal will be to detect mutations in Kras/Hras, FGFR3 (which are markers of low staged/low grade tumors) and p53 (which is a marker of high stage/high grade tumors) occurring at the 5% or lower level in the sample. A small set of matched tissues and urines will be provided by Dr. Kevin Loughlin, Professor of Surgery and Director of Urologic Research at Brigham and Women's Hospital (Boston, MA). During phase II, a molecular assay based on the results of Phase I will be further developed and evaluated in a clinical laboratory environment for detection of early bladder cancer. PUBLIC HEALTH RELEVANCE: There is an urgent need to develop an effective method for non-invasive detection of early stage urothelial bladder cancer, which is one of the most common cancers, and has one of the highest recurrence rates. The current standard methods for screening rely either on cytology, which has only 20% sensitivity, or on cystoscopy, which often results in false positives, and is invasive and expensive. The goal of this Phase I project is to explore the ability of a new molecular assay method based on mass spectrometry to detect early stages of urothelial bladder cancer by analyzing DNA obtained from urine for the presence of mutations in key genes associated with bladder cancer pathogenesis.