FORMS OF PROSTATE SPECIFIC ANTIGEN AND HK2 IN CANCER

癌症中前列腺特异性抗原和 HK2 的形式

• 批准号: 6150143
• 负责人: MARY J HEEB
• 金额: $ 24.15万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-05-01 至 2004-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6150143
• 关键词: antibody specificity; benign prostate hyperplasia; biomarker; clinical research; diagnosis design /evaluation; enzyme inhibitors; enzyme linked immunosorbent assay; human subject; kallikreins; macroglobulins; neoplasm /cancer diagnosis; prostate neoplasms; prostate specific antigen; protein isoforms; protein sequence; western blottings

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.

我们寻求扩展有前景的研究，表明前列腺特异性 抗原（PSA）和腺激肽释放酶（hK2）以多种形式存在于 前列腺癌（CaP）患者的血液和测量 PSAalpha2-巨球蛋白 (alpha2M) 形式的 PSA 比例 提高早期 CaP 和良性前列腺疾病之间的区分 增生（BPH）。 这将通过充分表征来证实 前瞻性患者样本。此外，我们还会衡量比例 PSA 在这些患者中以各种形式存在，并识别和测量 相关新型 CaP 标记 hK2 的形式。 我们发现hK2存在 在 CaP 等离子体中以多种形式存在，包括 hK2-alpha2M。 临床检测 未检测到 PSA-alpha2M，也没有针对 hK2 的临床检测。 我们 将确定哪种测量组合提供最大的 Cap 和 BPH 之间的区别。 由于 PSA 的形式也各不相同 早期与晚期 CaP，我们的检测也可能对预后有用。 这 目标是提供简单的 ELISA 检测，以减少侵入性、 诊断所需的昂贵程序和 ELISA 预后，帮助医生选择治疗 CaP 的积极程度 患者，或识别稍后将出现 CaP 的 BPH 患者。 它 假设 PSA 和 hK2 作为酶原分泌，被激活 需要时，然后通过蛋白酶抑制剂灭活。 这 合成 PSA 和 hK2 的细胞的病理生理学在 CaP 中存在差异 与 BPH 相比，并且可能在不同的 CaP 过程中。 PSA 之间的时间 或 hK2 的分泌和易位至血液可能不同，并且 他们在前列腺中接触的酶和抑制剂不同 来自血液中的那个。 因此，不同范围的 PSA 和 hK2 形成 与 BPH 相比，在 CaP 中以及在 CaP 的不同病程中都会发现。 为了实现我们的目标，我们将： 1. 确定 CaP 和 BPH 中 hK2 的形式 血浆并开发相关 hK2 形式的检测方法。 纯化 hK2 和 制备相关 hK2 的抗体和标准 hK2 抑制剂复合物 形式。 2. 前瞻性地从 300 个 CaP 中收集系列血样 患者并记录完整的病史。 收集预期样本 300 名未确诊的 PSA 升高患者。 3. 测量 PSA-alpha2M PSA-ACT、每个样本中的游离 PSA 和“总”PSA 并计算 每种 PSA 形式的比例。 同样测量相关的 HK2 表格。 4. 将各种形式的 PSA 和 hK2 的比例与 所有 CaP 患者的病程以确定预后价值 每次测量。 5. 关联各种形式的比例 PSA 和 hK2 对患者后续 CaP 或 BPH 诊断的影响 那些未被确诊的人。确定每个的特异性和敏感性 测量以确定其诊断价值。 技术将包括 蛋白质纯化、抗体和 ELISA 开发、免疫印迹、 酶活性测定和肽合成。