High-Throughput Screening and Validation of Molecular Targeted Chemoradiosensitizers

项目摘要 分子靶向药物与放射治疗（RT）的临床前和临床开发， 化学疗法对于改善患有难以治疗的癌症的患者的结果至关重要。 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应用。 放射肿瘤学试验主要影响因素包括临床前数据重现性差， 影响临床治疗敏感性的肿瘤间基因组异质性的临床前建模， 以及依赖肿瘤生长延迟而不是局部肿瘤控制（TCD 50）终点。有一个紧急的 需要克服这些障碍，以成功的靶向化学放射增敏剂的临床翻译。我们 建议建立一个综合的体外/体内管道，用于化学放射增敏靶向药物， 生物标志物相关性和适当验证，以便随后在患有 难以治疗的癌症的成功概率将大大高于过去。为了实现这一点， 我们提出三个具体目标。首先，通过利用美国基因组学协会独特的专业知识和资源， 癌症药物敏感性项目（马萨诸塞州总医院和英国威康信托桑格研究所） 和德国癌症研究中心/癌症联盟（DKFZ/DKTK）提供，我们将进行一个机器人 高通量筛选在基于细胞外基质（ECM）的3D格式中生长的癌细胞系，以更好地 模拟体内生长条件。我们建议筛选目前CTEP产品组合的一半（30种药物） 与在100个注释细胞系的初始组中的分次照射相结合，所述细胞系被选择来代表 临床相关的肿瘤间基因组变异。第二，我们提出了一个系统的、逐步的 验证/改进过程，以提名在动物中成功可能性最高的CTEP药物 试验.这将包括3D集落形成测定，增加疾病特异性化疗，确认 药效学反应、靶标/生物标志物验证以及患者来源的细胞系的整合， 异种移植第三，我们将评估最有前途的CTEP药物在体内的放化疗增敏作用 通过在有/无生物标志物的小鼠异种移植模型中依赖于TCD 50测定，并利用临床相关的 RT分次（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FOA的目标直接相关，我们将在最后 与联盟内外的调查人员合作，加快有针对性地 确定了具有更大功效的放化疗治疗。这项工作将得到以下方面的大力推动： DKFZ/DKTK调查人员在3D方面提供特殊专业知识的外国项目组成部分的整合 ECM和TCD 50检测的低预算成本，这将直接和间接地使NCI和 在美国的放射肿瘤学社区。通过整合广泛的药物基因组学专业知识， 和辐射生物学，跨学科的研究团队是独一无二的，以帮助改变临床前 发现具有伴随生物标志物的化学放射增敏靶向药物的方法。

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.