Noninvasive, Uniplex, Molecular, Pathomic Urinary Assay for Detection of Prostate Cancer

翻译后摘要：尽管在了解其基因组和分子基础的进展，前列腺癌（PCa） 仍然是美国男性中最常诊断的实体恶性肿瘤，也是癌症的第二大原因 死亡目前，诊断PCa的唯一方法是侵入性经直肠前列腺超声 活检大多数（>66%）活检显示良性病变，但患者发病率较高， 医疗保健美元因此，仍然需要一种简单且非侵入性的方法， 明确诊断PCa，确定它是侵袭性的，惰性的还是良性的，并帮助指导其管理。 随着分子生物学的发展，血液中PCa细胞的基因组分析受到了广泛的关注 和尿液。在两种这样的测定中，FDA批准的PCA 3和SelectMDx，在DRE后收集尿液，恶性肿瘤 细胞分离和PCa的多重基因组指纹特征。PCA 3测试虽然具有创新性， 广泛使用，主要是由于其灵敏度、特异性、阳性预测（PPV）和 阴性预测值（NPV）。此外，SelectMDx和一种经常提倡的血清的临床效用 4Kscore测试，仍不确定。 有大量文献表明VPAC受体在PCa细胞上表达。关于NIH/NCI 支持，我们设计并标记了VPAC特异性肽与Cu-64，使我们能够成像PCa成功地在 人类然后我们假设，在没有前列腺刺激的情况下，PCa患者排尿中脱落的PCa细胞， 通过DRE，可以通过靶向VPAC受体进行光学成像，使用标记有 荧光团。我们从PCa、BPH患者中收集的>250个去识别尿液样本的初步结果 和正常情况下，是令人鼓舞的（灵敏度>98%），并与我们的假设一致。在这次调查中 我们建议获得发展这种有前途的非侵入性技术所需的关键信息， 尿分析作为PCa诊断测试。我们的具体目标是1）确定灵敏度，特异性，阳性 预测值（PPV）和阴性预测值（NPV）的分子尿测定男性已知 PCa和阴性对照; 2）检查该测定法在治疗PCa患者中的有效性 既往PCa活检阴性，但PSA持续升高，计划进行TRUS活检; 3)为了确定是否a）恶性细胞占尿中脱落的总细胞的百分比，B）荧光 c）脱落的恶性细胞中的VPAC蛋白质量与恶性细胞周围的VPAC蛋白质强度相关， 4）评估这种分子尿检测在治疗中的作用， 积极监测的患者;和5）确定尿液储存是否需要防腐剂。这 简单、可靠和患者友好的单重测定将a）检测活动性、侵袭性或惰性PCa，B）挽救患者 c）减少不必要的活检和相关患者的数量 d）节省数百万美元的医疗费用。

Abstract: Despite the advances in understanding its genomic and molecular basis, prostate cancer (PCa) remains the most commonly diagnosed solid malignancy in men in the US and the second leading cause of cancer death. Currently, the only established method to diagnose PCa is an invasive transrectal ultrasound prostate biopsy. The majority (>66%) of biopsies show benign pathology at the expense of patient morbidity and healthcare dollars. There remains, therefore, an unmet need for a simple and non-invasive approach that will definitively diagnose PCa, determine if it is aggressive, indolent or benign and help guide its management. The era of molecular profiling has drawn much attention to genomic analysis of malignant PCa cells shed in blood and urine. In two such assays, the PCA3 and SelectMDx approved by FDA, urine is collected after DRE, malignant cells isolated and characterized by multiplex genomic finger prints of PCa. Although innovative, PCA3 test is not widely used, primarily due to its wide range (62%-94%) of sensitivity, specificity, positive predictive (PPV) and negative predictive value (NPV). Furthermore, the clinical utility of SelectMDx and a frequently advocated serum 4Kscore test, remains uncertain. There is robust literature demonstrating that VPAC receptors are expressed on PCa cells. With NIH/NCI support, we designed and labeled a VPAC specific peptide with Cu-64 that allowed us to image PCa successfully in humans. We then hypothesized that PCa cells, shed in voided urine of PCa patients, without prostate stimulation by DRE, can be imaged optically by targeting VPAC receptors, using the same peptide labeled with a fluorophore. Our preliminary results of >250 de-identified urine samples, collected from patients with PCa, BPH and normal cases, are encouraging (sensitivity >98%) and are consistent with our hypothesis. In this investigation we propose to obtain critical information that will be required for the development of this promising noninvasive urine assay as a PCa diagnostic test. Our specific aims are 1) To determine sensitivity, specificity, positive predictive (PPV), and negative predictive value (NPV) of the molecular urinary assay for men with known PCa and negative controls; 2) To examine the efficacy of the assay in the management of patients with previous negative biopsy for PCa but have persistently elevated PSA and are scheduled for TRUS biopsy; 3) To establish if a) the malignant cells as a percent of total cells shed in the urine, b) the fluorescence intensity around malignant cells, and c) the VPAC protein quantity in shed malignant cells correlate with the aggressiveness of the disease; 4) To assess the role of this molecular urine assay in the management patients on active surveillance; and 5) To determine if a preservative is required for urine storage. This simple, reliable and patient-friendly uniplex assay will a) detect active, aggressive or indolent PCa, b) save patients from over diagnosis and over treatment, c) reduce the number of unnecessary biopsies and associated patient morbidity, and d) save millions of healthcare dollars.