Development of a Colon Cancer GEMM-Derived Orthotopic Transplant Model for Drug Discovery and Validation
Development of a Colon Cancer GEMM-Derived Orthotopic Transplant Model for Drug Discovery and Validation
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DOI：
10.1158/1078-0432.ccr-12-2307
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发表时间：
2013-06-01
影响因子：
11.5
通讯作者：
Hung, Kenneth E.
中科院分区：
文献类型：
--
作者：
Martin, Eric S.;Belmont, Peter J.;Hung, Kenneth E.
目的：KRAS突变型结直肠癌（CRC）的有效治疗是一个关键的未满足的临床需求。先前，我们描述了适合于实验治疗的临床前评价的散发性Kras突变型和非突变型CRC的基因工程小鼠模型（GEMM）。为了加速药物的发现和验证，我们试图从GEMM Kras突变体和野生型肿瘤中获得低传代细胞系，用于体外筛选和移植到免疫活性小鼠的天然结肠环境中进行体内validation.Experimental Design：细胞系来源于在限定培养基条件下的Kras突变体和非突变体GEMM肿瘤。生长动力学，磷酸化蛋白质组，转录，药物敏感性和代谢进行了检查。将细胞系植入小鼠体内并监测体内肿瘤analysis.Results：Kras突变细胞系显示增殖增加，促分裂原活化蛋白激酶信号传导和磷酸肌醇-3激酶信号传导。微阵列分析确定了与人类CRC相关基因特征的显著重叠，包括KRAS突变和转移性CRC。进一步的分析显示了许多疾病相关的生物途径，包括葡萄糖代谢的富集。在体外和体内的功能评估验证了这一发现，并强调了致癌信号和有氧glycolyolysis.Conclusions的依赖KRAS突变CRC：我们已经成功地表征了一种新的GEMM衍生的原位移植模型的人KRAS突变CRC。该方法结合了使用重现人CRC的低传代细胞系的体外筛选能力和使用在免疫活性动物的结肠微环境中发展的细胞系来源的肿瘤的快速体内验证的潜力。总之，该平台是临床前CRC模型的一个明显进步，用于全面的药物发现和验证工作。(c)2013年AACR。
Purpose: Effective therapies for KRAS-mutant colorectal cancer (CRC) are a critical unmet clinical need. Previously, we described genetically engineered mouse models (GEMM) for sporadic Kras-mutant and non-mutant CRC suitable for preclinical evaluation of experimental therapeutics. To accelerate drug discovery and validation, we sought to derive low-passage cell lines from GEMM Kras-mutant and wild-type tumors for in vitro screening and transplantation into the native colonic environment of immunocompetent mice for in vivo validation.Experimental Design: Cell lines were derived from Kras-mutant and non-mutant GEMM tumors under defined media conditions. Growth kinetics, phosphoproteomes, transcriptomes, drug sensitivity, and metabolism were examined. Cell lines were implanted in mice and monitored for in vivo tumor analysis.Results: Kras-mutant cell lines displayed increased proliferation, mitogen-activated protein kinase signaling, and phosphoinositide-3 kinase signaling. Microarray analysis identified significant overlap with human CRC-related gene signatures, including KRAS-mutant and metastatic CRC. Further analyses revealed enrichment for numerous disease-relevant biologic pathways, including glucose metabolism. Functional assessment in vitro and in vivo validated this finding and highlighted the dependence of Kras-mutant CRC on oncogenic signaling and on aerobic glycolysis.Conclusions: We have successfully characterized a novel GEMM-derived orthotopic transplant model of human KRAS-mutant CRC. This approach combines in vitro screening capability using low-passage cell lines that recapitulate human CRC and potential for rapid in vivo validation using cell line-derived tumors that develop in the colonic microenvironment of immunocompetent animals. Taken together, this platform is a clear advancement in preclinical CRC models for comprehensive drug discovery and validation efforts. (c) 2013 AACR.