URINARY BIOMARKERS OF RENAL CELL CARCINOMAS

描述（由申请人提供）：肾癌通常是无声的，往往是致命的。仅在美国，2009年就有超过57,000例新的肾癌被诊断出来，近13000例死亡可归因于这种疾病。早期诊断，触发手术切除，通常是治愈的。身体体征和症状出现较晚，并且在随后诊断出肾癌时，它通常已经转移到淋巴结或邻近器官，并且进展超过治愈阶段，生存机会极小。偶然的早期发现可能发生在腹部成像（CT，MRI，超声）不相关的原因。尽管如此，目前还没有用于群体筛查和诊断肾癌的方法，特别是在治愈阶段。尚未满足的需求是一种简单、高通量、具有成本效益的肾癌筛查测试，其公共卫生意义是每年防止13，000多人死亡，特别是在高危人群中。拟议的研究将测试肾癌细胞表达和脱落或主动分泌蛋白质到尿液中的假设，这些蛋白质是标志性分子标志物（生物标志物），并且这些生物标志物是早期疾病指标，当肿瘤处于可治疗阶段时可检测到。[In在新闻出版物中，我们已经表明水通道蛋白1（AQP 1）和亲脂蛋白（ADFP）在肾癌患者的尿外泌体中显著增加，从而提供了这种疾病的第一个敏感和特异性生物标志物。因此，进一步假设尿外泌体，即由肾单位的所有部分释放到尿中的囊泡结构，将是[额外的新型生物标志物蛋白的发现工具，并将用作肾细胞肿瘤发生和转移的非侵入性活检]。总体目标是鉴定肾癌患者尿液外泌体中的蛋白质，这些蛋白质是这种疾病所特有的，并且可以开发成一种诊断测试来筛查[全尿]，以在早期可治愈阶段检测肾癌，并在术后随访患者复发。具体目标是发现阶段，以：1）鉴定肾癌患者的尿外泌体中的候选生物标志物蛋白，其是疾病的致病因素;开发阶段，以2）开发敏感和特异性ELISA，以测量Aim 1中鉴定的新型生物标志物蛋白的水平;以及验证阶段，以3）表征候选生物标志物蛋白，以确定其诊断灵敏度和特异性。独特的上调或下调蛋白质将通过尿液外泌体蛋白的无偏自上而下和监督自下而上质谱（LC/MS/MS）和蛋白质印迹蛋白质组学分析来鉴定。一旦通过质谱法在患者的训练集中鉴定出蛋白质，将开发出精确但经济的基于ELISA的免疫测定法，该方法可在同一天得到结果，用于患者验证集中的蛋白质定量[在全尿液和血清中]。总的来说，我们的研究将提高肾癌的诊断到足够早的阶段，以便在最大限度地提高未来肾功能的同时获得显著的治愈机会，[术后随访患者复发并确定未来选择性干预的目标]。 公共卫生相关性：肾癌通常是无声的，经常致命。仅在美国，2009年就有超过57，000例新的肾癌诊断，近13，000例死亡可归因于这种疾病。身体体征和症状出现较晚，并且在肾癌诊断时，它通常已经转移到淋巴结或邻近器官，并且进展超过治愈阶段，生存机会极小。尚未满足的需求是一种简单、高通量、具有成本效益的肾癌筛查和复发检测方法，其公共卫生意义是每年防止13，000多人死亡。我们的研究将在足够早的阶段改善肾癌的诊断，以允许显著的治愈机会，保留剩余的肾功能[并随访术后患者的复发和治疗，同时提供对肾癌肿瘤发生和转移的重要见解]。

DESCRIPTION (provided by applicant): Renal cancer is typically silent and frequently fatal. In the United States in 2009 alone over 57,000 new kidney cancers were diagnosed and almost 13, 000 deaths were attributable to this disease. Early diagnosis, triggering surgical excision, is usually curative. Physical signs and symptoms occur late, and by the time renal cancer is consequently diagnosed, it has usually metastasized to lymph nodes or adjacent organs and advanced beyond a curative stage with minimal chance for survival. Incidental early discovery may occur fortuitously during abdominal imaging (CT, MRI, ultrasound) for unrelated causes. Nonetheless, there is presently no method for population screening and diagnosis of renal cancer, particularly at a curative stage. The unmet need is a simple, high through-put, cost-effective test for kidney cancer screening, with the public health significance of preventing over 13,000 deaths annually especially in at risk populations. The proposed research will test the hypothesis that renal cancer cells express and shed or actively secrete proteins into the urine that are signature molecular markers (biomarkers), and that these biomarkers are an early disease indicator, detectable when the tumor is at a treatable stage. [In an in press publication, we have shown that aquaporin 1 (AQP1) and adipophilin (ADFP) are significantly increased in urinary exosomes of patients with kidney cancer thus providing the first sensitive and specific biomarkers of this disease.] Therefore, it is further hypothesized that urine exosomes, vesicular structures released into the urine by all parts of the nephron, will be a [discovery tool for additional novel biomarker proteins and will serve as a non-invasive biopsy of kidney cell oncogenesis and metastasis]. The overall objective is to identify proteins in the urine exosomes of kidney cancer patients which are unique to this disease, and which can be developed into a diagnostic test to screen [whole urine] to detect kidney cancer at an early curable stage and to follow patients post-operatively for recurrence. The specific aims are a discovery stage to: 1) Identify candidate biomarker proteins in the urine exosomes of patients with kidney cancer which are pathogneumonic of the disease; a development stage to 2) Develop sensitive and specific ELISAs to measure levels of novel biomarker proteins identified in Aim1; and a validation stage to 3) Characterize the candidate biomarker proteins to determine their diagnostic sensitivity and specificity. Unique up- or down-regulated proteins will be identified by unbiased top-down and supervised bottom-up mass spectral (LC/MS/MS) and Western blot proteomic analysis of the urine exosomal proteins. Once proteins are identified in training sets of patients by mass spectrometry, precise but economical ELISA-based immunologic assays that give same-day results will be developed, for protein quantification [in whole urine and serum] in validation sets of patients. Overall, our study will improve the diagnosis of kidney cancer to a stage early enough to allow a significant chance for cure while maximizing future kidney function, [follow patients post-operatively for recurrence and identify future targets for selective intervention]. PUBLIC HEALTH RELEVANCE: Renal cancer is typically silent and frequently fatal. In the United States alone in 2009 there were over 57,000 new kidney cancers diagnosed and almost 13,000 deaths attributable to this disease. Physical signs and symptoms occur late, and by the time renal cancer diagnosed, it has usually metastasized to lymph nodes or adjacent organs and advanced beyond a curative stage with minimal chance for survival. The unmet need is a simple, high through-put, cost-effective test for kidney cancer screening and recurrence with the public health significance of preventing over 13,000 deaths annually. Our study will improve the diagnosis of kidney cancer at a stage early enough to allow a significant chance for cure, preserve remaining kidney function [and follow post-operative patients for recurrence and treatment while providing important insight into kidney cancer oncogenesis and metastasis].