Detection of Prostate Cancer Genomic Signatures in Blood

描述（由申请人提供）：本授权申请涉及本FAO（RFA-OD-09-003）中描述的广泛挑战领域“03 -生物标志物发现和验证”和特定挑战主题“03-CA- 101 -用于癌症早期检测和治疗的指纹”。前列腺癌（PC）的诊断通常是在常规的前列腺特异性抗原（PSA）测定血液和/或发现前列腺内可触及的肿块（直肠检查后），可以通过解剖（US，CT）成像可视化。原发性肿块的性质通常通过经直肠、超声引导的前列腺穿刺活检后的组织病理学特征来确定。在临床上，这些肿瘤大多数位于前列腺，手术切除/放射治疗这些原发性肿瘤可以治愈。然而，对于许多发生转移的患者，尽管目前的治疗，结果是死亡。导致高死亡率的一个主要因素是我们无法在出现病理体征和症状之前，在最早/最可治愈的阶段识别男性中原发性和转移性隐匿性PC病变的存在。虽然目前的基于血液的测定（例如，PSA）在诊断和治疗监测中继续发挥一定作用，但其特异性、敏感性和准确性仍然不足。我们假设，困扰这种和其他癌症测定的假阳性和假阴性率是它们无意中依赖于从“健康”男性血液中获得的群体来源的平均基因组/蛋白质组特征谱的结果，即，基线/背景特征不是被测试个体的基因组成所特有的。最近，我们提出，从同一个体获得的吞噬细胞和非吞噬细胞WBC理想地适合于“肿瘤特异性”和“正常特异性”特征的简易鉴定和区分，因此，消除了由于内在个体间（例如，年龄、性别、种族背景、健康状况）和基因表达的时间变化。独特地，该方法不依赖于从“健康”对照获得的群体来源的平均特征谱和/或生物标志物值。该方法声称，吞噬WBC的DNA、RNA、蛋白质和/或脂质表达谱的分析及其与来自相同供体的非吞噬WBC的表达谱的比较将导致识别吞噬细胞内的肿瘤特异性特征（患者特异性信号），其在非吞噬细胞中不表达或最低程度地表达（患者特异性噪声）。我们在荷瘤小鼠和少数癌症患者中的初步研究完全支持我们的预期，并显示（i）多种癌基因和肿瘤特异性基因组标记在吞噬细胞内选择性获得/表达;（ii）这些基因在非吞噬细胞中不表达或最低限度表达;和（iii）该测定-迄今为止在所有情况下-区分（a）荷瘤小鼠和非荷瘤小鼠，和（B）癌症患者和正常献血者。在这项资助申请中提出的研究中，我们计划在从已知患有PC的男性中获得的血液样本中验证这种独特的方法，并证明新开发的新型血液测定法（i）可以区分已知患有PC的患者和健康个体（献血者），和（ii）将导致对PC特异性的基因组标记的鉴定，并且普遍预测隐匿性和/或复发性疾病的存在。我们相信，我们的研究-在PC患者中成功验证血液测定后-将导致发现基因阵列（用鉴定的基因组签名点样），其将（i）用于PC的常规检测/诊断，和（ii）用于鉴定疾病复发。因此，我们预计血液检测将对美国和全球根除PC的目标产生前所未有的高影响，并做出重大贡献。项目叙述：目前基于血液的细胞和分子检测的特异性、灵敏度、重现性和/或准确性不足。我们假设，困扰这些测定的假阳性率和假阴性率是它们依赖于从“健康”对照的血液获得的群体衍生的平均特征谱的结果，即，基线/背景特征不是被测试个体的基因组成所特有的。在这项挑战拨款申请中，我们描述了用于识别已知患有前列腺癌的男性WBC内肿瘤特异性签名的方法。

DESCRIPTION (provided by applicant): This grant application addresses the Broad Challenge Area "03 - Biomarker Discovery and Validation" as described within this FAO (RFA-OD-09-003) and the Specific Challenge Topic "03-CA- 101 - Fingerprints for the early detection and treatment of cancer". The diagnosis of prostate cancer (PC) is usually subsequent to routine prostate-specific antigen (PSA) determination in blood and/or discovery of a palpable mass within the prostate (following rectal examination) that may be visualized by anatomic (US, CT) imaging. The nature of the primary masses is generally affirmed by histopathologic characterization following an invasive transrectal, ultrasound-directed, prostatic biopsy. At presentation, the majority of these tumors are localized in the prostate and surgical removal/radiation of these primary tumors can be curative. However, for many of the patients who develop metastases, despite current therapies, the outcome is death. A principal factor contributing to the high mortality rate is our inability to identify the presence of primary and metastatic occult PC lesions in men at the earliest/most curable stage, prior to the manifestation of pathologic signs and symptoms. While current blood-based assays (e.g., PSA) continue to play some role in diagnosis and in treatment monitoring, their specificity, sensitivity, and accuracy continue to be inadequate. We postulate that the false-positive and false-negative rates which have plagued this and other cancer assays are a consequence of their inadvertent dependence on population-derived, average genomic/ proteomic signature profiles that are obtained from the blood of "healthy" men, i.e., the baseline/ background signature(s) is/are NOT specific to the genetic makeup of the individual being tested. Recently, we proposed that phagocytic and nonphagocytic WBC - obtained from the same individual - are ideally suited to the facile identification and differentiation of "tumor-specific" and "normal- specific" signatures and, therefore, the elimination of the "inequality of baseline" consequent to the intrinsic interindividual (e.g., age, gender, ethnic background, health status) and temporal variation in gene expression. Uniquely, the approach does not depend on population-derived average signature profiles and/or biomarker values obtained from "healthy" controls. The approach claims that the analysis of DNA, RNA, protein, and/or lipid expression profiles of phagocytic WBCs and their comparison with those from nonphagocytic WBCs of the same donor will lead to the identification of tumor-specific signatures within the phagocytic cells (patient-specific signal) that are not expressed or minimally expressed in the nonphagocytic cells (patient-specific noise). Our preliminary studies in tumor-bearing mice and in a few cancer patients fully support our expectations and show that (i) multiple oncogenes and tumor-specific genomic signatures are selectively acquired/expressed within phagocytic cells; (ii) these genes are not expressed or are minimally expressed in nonphagocytic cells; and (iii) the assay - thus far in all instances - differentiates (a) tumor-bearing mice from non-tumor bearing mice, and (b) cancer patients from normal blood donors. In the studies proposed in this grant application, we plan to validate this unique approach in blood samples obtained from men known to have PC and demonstrate that the newly-developed novel blood assay (i) can differentiate between patients known to have PC and healthy individuals (blood donors), and (ii) will lead to the identification of genomic signatures that are specific to PC and are universally predictive of the presence of occult and/or recurring disease. We trust that our studies - upon successful verification of the blood assay in PC patients - will lead to the discovery of a gene array (spotted with the identified genomic signatures) that will (i) be used for the routine detection/diagnosis of PC, and (ii) be employed to identify disease recurrence. Consequently, we expect the blood test to have an unprecedentedly high impact on, and contribute significantly to, the goal of eradicating PC in the USA and worldwide. PROJECT NARRATIVE: The specificity, sensitivity, reproducibility, and/or accuracy of current blood-based cellular and molecular assays are inadequate. We postulate that the false-positive and false-negative rates that have plagued these assays are a consequence of their dependence on population-derived, average signature profiles obtained from the blood of "healthy" controls, i.e., the baseline/background signature(s) is/are NOT specific to the genetic makeup of the individual being tested. In this Challenge grant application, we describe methods for the identification of tumor-specific signatures within the WBCs of men known to have prostate cancer.