Profiling Urine Glycosylation of PSA and other Glyco-Biomarkers in Prostate Cance

描述（由申请人提供）：尿液是泌尿生殖系统癌症生物标志物的一种容易获得但大量被丢弃的潜在来源。癌症的生物标志物出现在尿液中，甚至来自肺癌和乳腺癌等远程癌症。我们观察到显着水平的前列腺特异性抗原（PSA）和前列腺酸性磷酸酶（PAP）在尿液中的健康个体，与一些早期的研究结果相一致。虽然PSA是目前前列腺癌（PC）的生物标志物，但其可靠性受到个体之间基线水平变化以及各种可用试剂盒测量结果不一致的影响。迫切需要建立一种临床相关的测定方法，用于表征PC、良性前列腺增生（BPH）中产生的PSA（和PAP）以及健康细胞释放的PSA，该方法可以可靠地用于前列腺癌的早期检测、预后和监测。PSA和PAP都是糖基化的，糖基化的变化与癌症密切相关。最近的报告表明，癌症中PSA糖基化发生变化，但在不同的体液中测量（PC血清和健康精液），并且仅在一名患者中有效。我们开发了一种方法（GlycoMatic），用于分析富集蛋白中单个位点的糖基化。我们将改进分离方法以富集PSA、PAP和可能在临床样本中检测到的其他前列腺特异性糖蛋白。我们将测试该技术对标准品和标准品加标到尿样从健康人，获得可重复的测定的能力，和稳定性的糖蛋白时，储存在不太理想的条件下。为了消除可归因于遗传或表观遗传组成的任何变化，将使用60名正常志愿者的尿液确定个体PSA和PAP糖基化特征的变化。分析中将考虑到年龄和种族等变量的信息，这些信息将来自具有重要少数民族成分的人口。将测定PC和良性前列腺增生（BPH）患者（各60例）队列的PSA和PAP糖基化特征，并与健康志愿者的PSA和PAP糖基化特征进行比较。获得用于临床试验的糖基化谱的能力是一个新的领域。因此，将建立计算方法，以显示个体内、疾病的每个分类内的数据的可靠性，并与疾病状态、Gleason评分和PSA血清水平相关。这项研究开辟了新的领域，并将建立生物标志物蛋白糖基化谱在癌症早期检测，预测和监测中的临床适用性。它还将确定和区分由于种族而引起的任何变化。前列腺特异性抗原（PSA）是一种用于早期检测和监测前列腺癌（PC）的成熟测试。尽管如此，它的确定受到个体之间基础水平的变化以及各种可用试剂盒测量结果的不一致性的阻碍。癌症中产生的PSA被修饰（糖基化），这项有限的临床试验将建立差异，并确定这些差异是否足够特异，以清楚地区分PC产生的PSA和健康前列腺正常产生的PSA。前列腺酸性磷酸酶，前列腺癌的早期糖蛋白生物标志物，将进行类似的研究。将在尿液中进行分析，从而可能进行PC的自我检测。

DESCRIPTION (provided by applicant): Urine represents an easily available yet largely discarded potential source of biomarkers of urogenital cancers. Biomarkers of cancer appear in urine, even from remote cancers such as lung and breast. We observed significant levels of both prostate specific antigen (PSA) and prostatic acid phosphatase (PAP) in the urine from healthy individuals, consistent with some earlier findings. While PSA is the current biomarker for prostate cancer (PC), its reliability is hampered by variations in the baseline level between individuals, and the inconsistency in measurements made by the variety of kits available. There is an urgent need for establishing a clinically pertinent assay for the characterization of PSA (and PAP) produced in PC, benign prostatic hyperplasia (BPH) and that released by healthy cells, one that can be reliably used for the early detection, prognosis and monitoring of prostate cancer. Both PSA and PAP are glycosylated and changes in glycosylation go hand-in-hand with cancer. Recent reports have shown that changes occur in the glycosylation of PSA in cancer but were measured in different body fluids (serum for PC and seminal fluid for healthy), and effectively in only one patient. We have developed a method (GlycoMatic) for profiling glycosylation at individual sites in enriched proteins. We will refine isolation approaches to enrich the PSA, PAP and such other prostate-specific glycoproteins as may be detected in clinical samples. We will test the technique against standards and standards spiked into urine samples from healthy individuals, the ability to get reproducible determinations, and the stability of the glycoproteins when stored under less than ideal conditions. To eliminate any changes attributable to genetic or epigenetic make-up, variations in the individual PSA and PAP glycosylation profiles will be established using urines from 60 normal volunteers. Information on variables such as age and ethnicity will be factored into the analyses which will result from a population with a significant minority component. The PSA and PAP glycosylation profiles will be determined for a cohort of 60 (of each) PC and benign prostatic hyperplasia (BPH) patients and compared to that from the healthy volunteers. The ability to obtain glycosylation profiles for clinical trials is a new field. Thus, computational methods will be established that show the reliability of the data within individuals, within each classification of the disease, and correlated with disease status, Gleason score and PSA serum levels. This research breaks new ground and will establish the clinical applicability for the use of glycosylation profiles of biomarker proteins in the early detection, prediction and monitoring of cancer. It will also identify and distinguish any changes due to ethnicity. Prostate specific antigen (PSA) is a well-established test for the early detection and monitoring of prostate cancer (PC). Despite this, its determination is hampered by variation in the basal level between individuals, and the inconsistency in measurements made by the variety of kits available. PSA produced in cancer is modified (glycosylated) and this limited clinical trial will establish the differences, and determine if these are specific enough to clearly distinguish PC produced PSA from that normally produced by a healthy prostate. Prostatic acid phosphatase, an earlier glycoprotein biomarker of prostate cancer, will be similarly studied. Analyses will be performed in urine, leading to the possibility of a self-administered test for PC.