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

分子靶标核心开发技术和分析，使研究人员能够检查试验性新药(IND)对预期靶标及其相关途径的影响，以及指示治疗生物反应的相关生物标记物。我们在化验开发和验证、生物标记物分析和作用机制研究方面拥有专业知识。目前，我们正在设计和开发一些研究试剂的检测方法，以分析它们对细胞增殖、肿瘤血管生成和细胞凋亡的影响。1.自定义测试设计、开发和验证。我们开发、验证和实施基于电化学发光(ECL)的免疫分析方法用于临床标本的分析。尽管我们使用的ECL技术(罗氏诊断)是当今可用的最先进的临床免疫分析平台之一，但罗氏的任何标准测试都不适合我们的临床相关研究。因此，开发和定制这些分析方法以满足CCR临床研究人员的需求是至关重要的。我们的ECL平台非常适合这项持续的任务，因为它提供了高度的灵活性、稳定性和可靠性。另一个明显的优势是它的多重能力(即，能够使用有限数量的临床样本在一次分析中很好地确定总蛋白(t-)和磷酸蛋白(p-)的水平。由于临床样本可能差异很大，因此将这些样本正常化超过总蛋白浓度的能力对于生成具有统计学意义的患者样本数据至关重要。分子靶标核心具有开展这项工作的独特地位。2.目前和最近完成的生物标志物研究。目前，我们正在NCI-CCR使用大约10个临床方案。对于这些临床试验中的许多，我们帮助设计、开发、验证和实施用于相关分析研究的定制生物标记物分析。索拉非尼治疗雄激素非依赖性前列腺癌的II期临床试验(Dahut WL，et al.克莱恩。癌症研究14：209-14,2008)我们的实验室用调查剂索拉非尼进行了化验验证，并对骨髓活检标本进行了化验，以确定其对靶向MAP激酶途径的影响。在另一项研究联合靶向治疗(索拉非尼和贝伐单抗)的临床试验中，我们团队开发并分析了在药物试验期间从患者血浆中获得的血管生成因子血管内皮生长因子(VEGF)和细胞因子，包括白介素6(IL6)和白介素8(IL8)(Azad NS等人)。J·克莱恩。奥科尔。26：3709-14,2008)。在另一项试验中，我们测量了服用雷帕霉素后骨肉瘤活检组织和外周血单个核细胞(PBMC)S6RP信号的相对变化。我们的数据显示，PBMC中的p-S6RP在统计学上有非常显著的降低。P/t-S6RP比值从服药前的25%变化到服药后的0.5%，p<0.001，从服药第2天到研究结束。我们的结果进一步表明，肿瘤活检组织有很高的变异性。雷帕霉素对S6RP通路有统计学意义的影响只能用p/t-S6RP比值来证明，我们的双链测定法能够同时定量测定p-S6RP和t-S6RP(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).