Urinary MMP activity to detect renal cell carcinoma

描述(申请人提供)：被诊断为肾细胞癌(RCC)--最常见的肾癌--患者的存活率极低。一个主要原因是早期肾癌通常没有症状，导致已经出现转移性疾病的患者比例很高。目前尚无临床相关标记物可用于早期肾癌的检测。理想情况下，肾细胞癌的筛查试验应该是非侵入性的，并且应该有可能发展成具有成本效益的高通量试验。肾细胞癌特异的尿液标志物可以满足这些标准。我们的初步结果表明，肾癌患者的尿液中含有高水平的基质金属蛋白酶(MMPs)活性，这会导致正常排泄的细胞外基质(ECM)蛋白的降解。初步分析，尿ECM蛋白缺失检测肾癌的灵敏度为95%(21/22)，特异度为95%(21/22)。重要的是，所有早期病例都得到了正确识别。本项目的总体目标是建立一种基于尿MMPs或MMPs活性检测的快速筛查方法，并测试其在肾癌检测中的临床应用价值。首先，将使用特定的抗体或亲和层析和测序来鉴定分泌的基质金属蛋白酶(S)。其次，基于荧光底物的降解和/或MMPs的免疫学检测，将建立微量平板筛选试验。第三，使用较大的肾癌患者和对照人群，将确定所开发的筛查试验(S)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.