Urinary MMP activity to detect renal cell carcinoma

描述（由申请人提供）：肾细胞癌（RCC）-最常见的肾癌类型-患者的生存率极低。一个主要原因是，早期RCC通常是无症状的，导致高频率的患者已经存在转移性疾病。没有临床相关的标志物可用于检测早期RCC。理想情况下，RCC的筛选测定应该是非侵入性的，并且应该有可能发展成具有成本效益的高通量测定。RCC特异性尿标记物可满足这些标准。我们的初步研究结果表明，肾细胞癌患者的尿液中含有增加的基质金属蛋白酶（MMP）活性水平，导致正常分泌的细胞外基质（ECM）蛋白的降解。在初步分析中，检测到尿ECM蛋白的缺失使得可以以95%（21/22）的灵敏度和95%（21/22）的特异性检测RCC。重要的是，所有早期病例都得到了正确识别。本课题的主要目的是建立一种基于尿基质金属蛋白酶或基质金属蛋白酶活性检测的肾细胞癌快速筛查方法，并验证其在肾细胞癌检测中的临床应用价值。首先，将使用特异性抗体或通过亲和层析和测序鉴定分泌的MMP。其次，将基于荧光底物的降解和/或MMP的免疫学检测开发微量滴定板筛选试验。第三，使用较大的RCC患者和对照群体，将确定所开发的筛选测定的灵敏度和特异性。

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.