High-Throughput Screening and Validation of Molecular Targeted Chemoradiosensitizers

项目总结 放射治疗分子靶向药物的临床前和临床研究进展 化疗对于改善难治性癌症患者的预后至关重要。 然而，大量的临床前药物/RT研究并没有转化为足够数量的成功 放射肿瘤学试验。主要原因包括临床前数据的可重复性差、不足 临床上影响治疗敏感性的肿瘤间基因组异质性的临床前模型， 依赖肿瘤生长延迟而不是局部肿瘤控制(TCD50)终点。存在着一个紧急情况 需要克服这些障碍，才能成功地将靶向化疗放射增敏剂转化为临床药物。我们 建议建立一个一体化的体外/体内管道，用于对以下靶向药物进行化学放射增敏 生物标记物相关并适当验证，因此后续的临床药物/RT试验 难治性癌症的成功几率将大大高于过去。为了实现这一目标， 我们提出了三个具体目标。首先，通过利用基因组学的独特专业知识和资源 癌症药物敏感性项目(马萨诸塞州总医院和英国惠康信托桑格研究所) 和德国癌症研究中心/癌症联盟(DKFZ/DKTK)的报价，我们将进行机器人 高通量筛选以细胞外基质(ECM)为基础的3D格式生长的癌细胞系以更好地 模拟体内生长条件。我们建议对目前CTEP组合中的大约一半(30种药物)进行筛选 结合对100个注释细胞系的初始面板的分次照射，这些细胞系被选为代表 临床相关的肿瘤间基因组变异。其次，我们提出了系统的、循序渐进的 验证/改进过程，以提名在动物身上具有最高成功可能性的CTEP药物 测试。这将包括3D集落形成分析，增加疾病特异性化疗，确认 药效学反应、靶点/生物标记物验证以及患者来源的细胞系和 异种移植物。第三，我们将在体内评估最有希望的CTEP药物的化学和放射增敏作用。 在有/无生物标志物的小鼠异种移植模型中依赖TCD50检测，并利用临床相关 RT分级(30次/6周)。这些实验将再次利用特殊的DKFZ/DKTK能力。 拟议的研究与基本工作协议的目标直接相关，我们将在结束时 与财团内外的调查人员合作，加快目标 确定了疗效更好的放化疗方法。这项工作将得到以下方面的大力推动 整合外国项目组成部分，由DKFZ/DKTK调查员提供3D方面的专门知识 ECM和TCD50以低预算成本进行分析，这将直接和不成比例地使NCI和 美国的放射肿瘤学社区。通过整合药物基因组学方面的广泛专业知识 和放射生物学，跨学科的研究团队独一无二地准备帮助转变临床前 具有伴随生物标志物的靶向药物化学放射增敏的发现过程。

PROJECT SUMMARY Preclinical and clinical development of molecular targeted drugs with radiation therapy (RT) and chemotherapy are critically important for improving the outcomes of patients with hard-to-treat cancers. However, a huge body of preclinical drug/RT studies has not translated into an adequate number of successful radiation oncology trials. Major contributing factors include poor reproducibility of preclinical data, insufficient preclinical modeling of inter-tumoral genomic heterogeneity that influences treatment sensitivity in the clinic, and reliance on tumor growth delay instead of local tumor control (TCD50) endpoints. There exists an urgent need to overcome these barriers to successful clinical translation of targeted chemoradiosensitizers. We propose to establish an integrated in-vitro/in-vivo pipeline for chemoradiosensitizing targeted drugs that are biomarker-correlated and appropriately validated, so that subsequent clinical drug/RT trials in patients with hard-to-treat cancers will have a substantially higher probability of success than in the past. To achieve this, we propose 3 Specific Aims. First, by leveraging the unique expertise and resources that the Genomics of Drug Sensitivity in Cancer project (Massachusetts General Hospital & UK Wellcome Trust Sanger Institute) and the German Cancer Research Center/Cancer Consortium (DKFZ/DKTK) offer, we will conduct a robotic high-throughput screen of cancer cell lines grown in an extracellular-matrix (ECM) based 3D format to better mimic in-vivo growth conditions. We propose to screen about half of the current CTEP portfolio (30 drugs) combined with fractionated irradiation across an initial panel of 100 annotated cell lines selected to represent clinically relevant inter-tumoral genomic variation. Second, we propose a systematic and stepwise validation/refinement process to nominate CTEP drugs that have the highest likelihood to succeed in animal testing. This will include 3D colony formation assays, addition of disease-specific chemotherapy, confirmation of pharmacodynamic responses, target/biomarker validation, and integration of patient-derived cell lines and xenografts. Third, we will evaluate the chemoradiosensitizing effects of the most promising CTEP drugs in-vivo by relying on TCD50 assays in mouse xenograft models with/without biomarker and utilizing clinically relevant RT fractionation (30 fractions/6 weeks). These experiments again will leverage special DKFZ/DKTK capability. The proposed studies are directly relevant to the objectives of the underlying FOA, as we will, in close collaboration with investigators within and outside the consortium, accelerate the pace at which targeted chemoradiation treatments with greater efficacy are identified. This undertaking will be greatly facilitated by the integration of a foreign project component where DKFZ/DKTK investigators contribute special expertise in 3D ECM and TCD50 assays at low budget cost, which will directly and disproportionally benefit the NCI and the radiation oncology community in the United States. By integrating extensive expertise in pharmacogenomics and radiation biology, the inter-disciplinary investigator team is uniquely poised to help transform the preclinical discovery process for chemoradiosensitizing targeted drugs with accompanying biomarkers.