FORMS OF PROSTATE SPECIFIC ANTIGEN AND HK2 IN CANCER

我们寻求扩展有希望的研究，表明前列腺特异性 抗原（PSA）和腺激肽释放酶（hK 2）以几种形式出现在 前列腺癌（CaP）患者的血液和测量 PSA α 2-巨球蛋白（α 2 M）形式的PSA比例 提高早期前列腺增生与良性前列腺的鉴别能力 增生（BPH）。 这将通过充分表征的 前瞻性患者样本。此外，我们还将衡量 PSA在这些患者中的各种形式，并确定和测量 相关的新型CaP标记物hK 2的形式。 我们发现hK 2存在于 以几种形式存在于CaP血浆中，包括hK 2-α 2 M。 临床测定 不能检测PSA-α 2 M，也没有hK 2的临床试验。 我们 将确定哪种测量组合提供最大的 区分CaP和BPH。 由于PSA的形式也不同， 早期与晚期CaP相比，我们的测定也可用于预后。 的 目的是提供简单的ELISA测定， 诊断所需的昂贵程序和 预后，以帮助医生选择如何积极治疗CaP 患者，或确定BPH患者谁将发展CaP以后。 它 假设PSA和hK 2作为酶原分泌，活化， 当需要时，然后用蛋白酶抑制剂灭活。 的 合成PSA和hK 2的细胞的病理生理学在CaP中变化 vs. BPH和可能在不同的CaP过程中。 PSA之间的时间 或hK 2分泌和转运到血液中可能不同， 它们在前列腺中接触的酶和抑制剂 从血液中。 因此，PSA和hK 2形成不同的谱 将在CaP中与BPH相比，以及在CaP的不同过程中发现。 为了实现我们的目标，我们将：1。识别CaP和BPH中的hK 2形式 血浆和开发相关hK 2形式的测定。 纯化hK 2， 制备相关hK 2的抗体和标准hK 2-抑制剂复合物 forms. 2.普罗维登斯从300名CaP中收集系列血液样本 病人和文件完整的历史。 收集前瞻性样本， 300例未确诊的PSA升高患者。 3.测量PSA-α 2 M 每个样本中的PSA-ACT、游离PSA和“总”PSA，并计算 每种PSA形式的比例。 类似地测量相关的hK 2形式。 4.将各种形式的PSA和hK 2的比例与 所有CaP患者的病程，以确定预后价值 每一次测量。5.将各种形式的 PSA和hK 2对前列腺增生或前列腺增生的诊断价值 未确诊的人确定每种方法的特异性和灵敏度 测量以确定其诊断价值。 技术将包括 蛋白纯化，抗体和ELISA开发，免疫印迹， 酶活性测定和肽合成。

We seek to extend promising studies showing that prostate specific antigen (PSA) and glandular kallikrein (hK2) occur in several forms in blood of prostate cancer (CaP) patients and that measurement of the proportion of PSA in the form of PSAalpha2-macroglobulin (alpha2M) improves the discrimination between early CaP and benign prostatic hyperplasia (BPH). This will be confirmed with well-characterized prospective patient samples. Moreover, we will measure the proportion of PSA in its various forms in these patients, and identify and measure forms of the related novel CaP marker, hK2. We found that hK2 exists in several forms in CaP plasmas, including hK2-alpha2M. Clinical assays do not detect PSA-alpha2M and there are no clinical assays for hK2. We will determine which combination of measurements provides the greatest discrimination between CaP and BPH. Since forms of PSA also vary in early vs. advanced CaP, our assays may also be useful in prognosis. The goal is to provide simple ELISA assays that will reduce the invasive, expensive procedures that are needed for diagnosis and ELISAs that are prognostic, to help physicians choose how aggressively to treat CaP patients, or to identify BPH patients who will develop CaP later. It is hypothesized that PSA and hK2 are secreted as zymogens, activated when needed, and then inactivated by protease inhibitors. The pathophysiology of cells that synthesize PSA and hK2 vary in CaP vs. BPH and possibly in different courses of CaP. The time between PSA or hK2 secretion and translocation to the blood may differ, and the enzymes and inhibitors to which they are exposed in the prostate differs from that in blood. Thus, a different spectrum of PSA and hK2 forms will be found in CaP compared to BPH, and in different courses of CaP. To meet our goals, we will: 1. Identify forms of hK2 in CaP and BPH plasmas and develop assays for relevant hK2 forms. Purify hK2 and prepare antibodies and standard hK2-inhibitor complexes for relevant hK2 forms. 2. Prospectively collect serial blood samples from 300 CaP patients and document full histories. Collect prospective samples from 300 undiagnosed patients with elevated PSA. 3. Measure PSA-alpha2M PSA-ACT, free PSA and "total" PSA in each sample and calculate the proportions of each PSA form. Similarly measure relevant hK2 forms. 4. Correlate the proportions of various forms of PSA and hK2 to the course of disease in all CaP patients to determine the prognostic value of each measurement. 5. Correlate the proportions of various forms of PSA and hK2 to the subsequent diagnosis of CaP or BPH in the patients who were undiagnosed. Determine the specificity and sensitivity of each measurement to determine its diagnostic value. Techniques will include protein purification, antibody and ELISA development, immunoblotting, enzyme activity assays, and peptide synthesis.