Urinary MMP activity to detect renal cell carcinoma

描述（由申请人提供）：肾细胞癌（RCC）是肾癌中最常见的一种，诊断为肾细胞癌的患者生存率极低。一个主要原因是早期肾细胞癌通常无症状，导致患者出现转移性疾病的频率很高。没有临床相关的标志物可用于检测早期肾细胞癌。理想情况下，RCC的筛选试验应该是非侵入性的，并且应该有可能发展成为一种具有成本效益的高通量试验。rcc特异性尿液标志物可能符合这些标准。我们的初步结果表明，肾癌患者尿液中基质金属蛋白酶（MMP）活性水平升高，导致正常排泄的细胞外基质（ECM）蛋白降解。在初步分析中，检测尿ECM蛋白缺失使得检测RCC的灵敏度为95%(21/22)，特异性为95%（21/22）。重要的是，所有早期病例都被正确识别。该项目的总体目标是开发一种基于尿液MMPs或MMP活性检测的快速筛选方法，并测试其在检测RCC方面的临床实用性。首先，将使用特异性抗体或亲和层析和测序来鉴定排出的MMP。其次，基于荧光底物的降解和/或MMPs的免疫学检测，将开发微滴度板筛选试验。第三，使用更大的RCC患者和对照人群，将确定开发的筛选试验的敏感性和特异性。

DESCRIPTION (provided by applicant): The survival rates for patients diagnosed with renal cell carcinoma (RCC) -the most common type of kidney cancer- are extremely low. A major reason is that early-stage RCC is usually asymptomatic leading to a high frequency of patients that present already with metastatic disease. No clinically relevant marker is available that would allow the detection of early-stage RCC. Ideally, a screening assay for RCC should be non-invasive, and should be possible to be developed into a cost-effective, high-throughput assay. An RCC-specific urinary marker may fulfill these criteria. Our preliminary results indicate that urine from RCC patients contains increased levels of matrix metalloproteinase (MMP) activity which causes the degradation of normally excreted extracellular matrix (ECM) proteins. In a preliminary analysis, the detection of the absence of urinary ECM proteins has allowed the detection of RCC with a sensitivity of 95% (21/22) and specificity of 95% (21/22). Importantly, all early-stage cases were correctly identified. The over-all goal of this project is to develop a rapid screening assay based on the detection of urinary MMPs or MMP activity, and to test its clinical usefulness for the detection of RCC. First, the excreted MMP(s) will be identified using specific antibodies or by affinitychromatography and sequencing. Second, a micro-titer plate screening assay will be developed based on the degradation of fluorogenic substrates and/or on the immunological detection of MMPs. Third, using larger RCC patient and control populations, the sensitivity and specificity of the developed screening assay(s) will be determined.