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

摘要：尽管对前列腺癌(PCa)的基因组和分子基础有了深入的了解，但前列腺癌(PCa) 仍然是美国男性最常见的被诊断为实体恶性肿瘤，是第二大致癌原因 死亡。目前，唯一确定的诊断前列腺癌的方法是经直肠超声前列腺癌。 活组织检查。大多数(&gt；66%)活检显示良性病理，但患者的发病率和 医疗保健美元。因此，对一种简单和非侵入性方法的需求仍然没有得到满足，这种方法将 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。 分子图谱时代已经引起了人们对血液中恶性PCa细胞的基因组分析的极大关注 还有尿液。在FDA批准的PCA3和SelectMDx两种这样的检测中，尿液是在DRE之后收集的，恶性 分离细胞，并用多重基因组指纹图谱进行鉴定。尽管PCA3考试是创新的，但它并不 广泛应用，主要是由于其灵敏度、特异度、阳性预测值(PPV)和 阴性预测值(NPV)。此外，SelectMDx和一种经常提倡的血清的临床应用 4KScore测试，仍不确定。 有可靠的文献表明VPAC受体在前列腺癌细胞上表达。与NIH/NCI合作 支持下，我们设计并标记了一种Vpac特异性多肽--铜-，使我们能够成功地在 人类。然后我们假设前列腺癌患者排尿中的前列腺癌细胞在没有前列腺刺激的情况下脱落。 通过DRE，可以通过靶向VPAC受体进行光学成像，使用与 荧光团。我们对收集自前列腺癌、前列腺增生症患者的250份未鉴定尿样的初步结果 和正常情况下，是令人鼓舞的(敏感性&gt；98%)，并与我们的假设一致。在这次调查中 我们建议获得开发这种有前景的非侵入性技术所需的关键信息 尿液化验作为前列腺癌诊断试验。我们的具体目标是：1)确定敏感性、特异性、阳性 已知男性尿液分子检测的预测价值(PPV)和阴性预测价值(NPV) Pca和阴性对照；2)检验该检测方法在治疗慢性前列腺癌患者中的有效性 先前的前列腺癌活检为阴性，但PSA持续升高，并计划进行TRUS活检； 3)确定a)恶性细胞占总细胞的百分比是否在尿液中排出，b)荧光 C)脱落的恶性细胞中VPAC蛋白的含量与 疾病的侵袭性；4)评估这种分子尿分析在治疗中的作用。 对患者进行积极监测；以及5)确定是否需要防腐剂来储存尿液。这 简单、可靠和患者友好的Uniplex检测将a)检测积极的、侵略性的或惰性的PCA，b)挽救患者 由于过度诊断和过度治疗，c)减少不必要的活检和相关患者的数量 发病率，以及d)节省数百万医疗保健费用。

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.