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

背景：靶向抗癌治疗的发展需要研究药物对预期靶点和途径的影响，并发现与反应相关的标记物。因此，我们致力于检测药物对途径特异性生物标记物的影响，并发现将在相关研究中实施的响应性生物标记物。技术、化验设计、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。这是当今最灵敏、最定量的免疫分析技术平台。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。它能够用有限数量的临床标本进行多重分析，在一次检测中很好地确定总蛋白和磷酸蛋白的水平。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。我们已经成功地将这项技术用于免疫分析，检测NCI临床方案中ERK、AKT通路、mTOR抑制、血管生成因子以及各种细胞因子的变化。我们目前是NCI唯一在这些平台上提供服务和协作的化验实验室。我们不断地挑战极限，提出能够满足临床试验中未得到满足的需求的新的分析方法。目前和最近完成的生物标记物研究。目前，我们在NCI-CCR参与了10项临床方案。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。以下是一些例子：1)对于索拉非尼治疗雄激素非依赖性前列腺癌的II期临床试验(PI，William Dahut)，我们实验室使用调查剂索拉非尼进行了试验验证，并对骨髓活检标本进行了分析，以确定其对靶向MAP激酶途径的影响(Dahut WL，等)。克莱恩。癌症研究报告14：209-14,2008)。2)在另一项研究卵巢癌联合靶向治疗的临床试验(PI，Elise Kohn)中，我们团队开发并分析了在整个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)。3)在另一项雷帕霉素试验(PI、Chand Khanna)中，我们检测了给药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比值从用药前的25%变化到用药后的0.5%，P&lt；0.001，样本采集从第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通路的显著影响只能用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(提交稿件)。支持药物开发的临床前研究。我们致力于广泛的临床前研究，以支持NCI的临床研究。例如：1)与Lee Helman博士合作，发现抗横纹肌肉瘤IGF1R抗体的预测生物标记物。2)Giuseppe Giaccon博士抗IGF1R治疗胸腺癌的可行性研究。3)与DCTD的Stephen Yoo博士合作，评估TORC1/2抑制剂Palomid 529与放射联合治疗头颈癌的效果。4)HIF1a抑制物高灵敏度检测方法的建立和评价。总之，对药效生物标记物和预测性生物标记物的生物标记物分析是早期抗癌药物开发的重要组成部分。我们在化验开发、验证和实施方面拥有独特的能力，我们积极参与NCI的临床试验。

Background: The development of targeted anti-cancer therapies requires the demonstration of the effects of the investigation agents on the intended target and pathway and the discovery of the markers that can be associated with the response. Thus, we are engaged in detecting the effects of agents on pathway-specific biomarkers and in discovering response biomarkers to be implemented in correlative studies. I. Technology, Assay Design, Development, and Validation. We develop, validate, and implement assays for clinical specimens using electrochemiluminescence (ECL)-based immunoassays. This is the most sensitive and quantitative immunoassay technology platform today. The ECL platform is well suited for this ongoing task because it offers a high degree of flexibility, stability and reliability. It is capable of multiplex analysis to determine the levels of total and phospho-proteins in a single assay well using a limited amount 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. We have successfully utilized the technology for immunoassays that detect changes in ERK, AKT pathways, mTOR inhibition, angiogenic factors, as well as a variety of cytokines in clinical protocols at NCI. We are currently the only lab at NCI provides services and collaboration with assays on these platform. We are constantly pushing the envelope to come up with new assays capable of meeting the unmet needs in clinical trials. II. Current and Recently Completed Biomarker Studies. Currently, we are engaged with 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. The following are some examples: 1) For a phase II clinical trial of sorafenib in androgen-independent prostate cancer (PI, William Dahut), our laboratory performed assay validation with the investigational agent sorafenib and implemented assays with bone marrow biopsy specimens to determine its effects on the targeted MAP kinase pathway (Dahut WL, et al. Clin. Cancer Res. 14: 209-14, 2008). 2) For another clinical trial that investigated a combination targeted therapy against ovarian cancer (PI, Elise Kohn), 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). 3) In another rapamycin trial (PI, Chand Khanna), 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<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 (manuscript submitted). III. Preclinical investigations in support of drug development. We are engaged with a wide range of preclinical investigations in support of clinical studies at NCI. Some examples are: 1) Discovering the predictive biomarker for anti-IGF1R antibodies against rhabdomyosarcoma in collaboration with Dr. Lee Helman. 2) Feasibility studies for anti-IGF1R therapy in thymic cancers with Dr. Giuseppe Giaccone. 3) Evaluation of TORC1/2 inhibitor Palomid 529 in combination with radiation against head and neck cancers in collaboration with Dr. Stephen Yoo of DCTD. 4) Development and evaluation of highly sensitive assay HIF1a assay as a biomarker for HIF1a inhibitors. In summary, biomarker analysis for pharmacodynamic biomarkers and predictive biomarkers is an essential part of early stage cancer drug development. We have unique capabilities in assay development, validation and implementation, and we actively engage clinical trials at NCI.