Design, Develop, Validate, and Implement Biomarkers for Clinical Investigations

分子靶点核心开发的技术和检测方法，使研究人员能够 检查研究性新药（IND）对预期靶点的影响及其 相关的途径，以及相关的生物标志物，指示生物学 对治疗的反应。我们在检测开发和验证、生物标志物 分析和作用机制研究。目前，我们正在设计和开发 许多研究药物，以分析其对细胞增殖，肿瘤， 血管生成和凋亡。1.定制检测设计、开发和验证。我们发展， 使用电化学发光验证和实施临床标本的检测 （ECL）的免疫测定。虽然我们使用的ECL技术（罗氏诊断）是一种 当今最先进的临床免疫分析平台，没有一个标准的 罗氏公司的检测试剂盒适用于我们的临床相关研究。因此，必须 开发和定制这些检测试剂盒，以专门满足CCR临床需求 investigators.我们的ECL平台非常适合这一持续的任务，因为它提供了一个高 灵活性、稳定性和可靠性。另一个明显的优势是它的多元化 能力（即，测定总蛋白（t-）和磷蛋白（p-）水平的能力 使用有限数量的临床样本在单个测定孔中进行。因为临床样本 可能会有很大的不同，除了总蛋白外， 浓度对于用患者标本生成统计学显著数据至关重要。 Molecular Targets Core是开展这项工作的唯一定位。2.电流和 最近完成的生物标志物研究。目前，我们正在与大约10个临床 NCI-CCR的协议对于许多这样的临床试验，我们帮助设计，开发， 验证和实施定制的生物标志物测定，用于相关分析研究。用于 索拉非尼在雄激素非依赖性前列腺癌中的II期临床试验（Dahut WL，et al. 14：209-14，2008），我们的实验室用以下方法进行了测定验证： 研究药物索拉非尼，并使用骨髓活检标本进行测定， 确定其对靶向MAP激酶途径的影响。另一项临床试验， 研究了联合靶向治疗（索拉非尼和贝伐单抗），我们的小组开发了 检测并分析血管生成因子血管内皮生长因子 （VEGF）和细胞因子，包括来自血浆的白细胞介素-6（IL 6）和白细胞介素-8（IL 8）。 在整个药物试验期间从患者获得（Azad NS，et al. J. Clin. Oncol. 26：3709-14，2008）。在另一项试验中，我们测量了S6 RP相对变化的变化， 来自骨肉瘤活检和外周血单核细胞的信号传导 在施用雷帕霉素后，在PBMC中观察到细胞凋亡。我们的数据显示， PBMC中p-S6 RP的显著降低。p/t-S6 RP比值从给药前的25%变为0.5% 给药后，p <0.001，从第2天至研究结束采集样本。我们 结果进一步显示肿瘤活检中存在高度的变异。的 雷帕霉素对S6 RP途径的统计学显著作用只能证明为 使用我们的能够定量测量p-S6 RP和p-S6 RP的双重测定， 相同试验孔中的t-S6 RP（未发表结果）。

The Molecular Targets Core develops technologies and assays that allow investigators to examine the effects of investigational new drugs (INDs) on intended targets and their associated pathways, as well as relevant biomarkers that are indicative of biological responses to the treatment. We have expertise in assay development and validation, biomarker analysis, and mechanism of action studies. Currently, we are designing and developing assays for a number of investigational agents to analyze their effects on cell proliferation, tumor angiogenesis, and apoptosis. 1. Custom Assay Design, Development, and Validation. We develop, validate, and implement assays for clinical specimens using electrochemiluminescence (ECL)-based immunoassays. Although the ECL technology (Roche Diagnostics) that we use is one of the most advanced clinical immunoassay platforms available today, none of the standard tests from Roche is suited for our clinical correlative studies. Therefore it is essential to develop and customize these assays to be specifically tailored to the needs of CCR clinical investigators. Our ECL platform is well suited for this ongoing task because it offers a high degree of flexibility, stability and reliability. Another distinct advantage is its multiplex capability (i.e., the ability to determine the levels of total (t-) and phospho-proteins (p-) in a single assay well using a limited number of clinical specimens. Because clinical samples may vary dramatically, the ability to normalize these samples beyond total protein concentration is critical in generating statistically significant data with patient specimens. The Molecular Targets Core is uniquely positioned to carry out this work. 2. Current and Recently Completed Biomarker Studies. Currently, we are working with about 10 clinical protocols at NCI-CCR. For many of these clinical trials, we helped to design, develop, validate, and implement customized biomarker assays for correlative analytical studies. For a phase II clinical trial of sorafenib in androgen-independent prostate cancer (Dahut WL, et al. Clin. Cancer Res. 14: 209-14, 2008) our laboratory performed assay validation with the investigational agent sorafenib and implemented assays with bone marrow biopsy specimens to determine the its effects on the targeted MAP kinase pathway. For another clinical trial that investigated a combination targeted therapy (sorafenib and bevacizumab), our group developed assays and performed the analysis of angiogenic factor vascular endothelial growth factor (VEGF) and cytokines, including interleukin-6 (IL6) and interleukin-8 (IL8) from plasma obtained from the patients throughout the drug trial period (Azad NS, et al. J. Clin. Oncol. 26: 3709-14, 2008). In another trial, we measured the changes in the relative changes of S6RP signaling from both osteosarcoma biopsies and peripheral blood mononuclear cells (PBMC)following the administration of rapamycin. Our data showed a highly statistically significant reduction of p-S6RP in PBMC. The p/t-S6RP ratio changed from 25% pre-drug to 0.5% post-drug with p less than 0.001, with samples obtained from day 2 to the end of the study. Our results further showed that there is a high degree of variation in tumor biopsies. The statistically significant effect of rapamycin on the S6RP pathway can only be demonstrated as p/t-S6RP ratio using our duplex assay capable of quantitatively measuring both p-S6RP and t-S6RP in the same assay well (unpublished results).