Di Nicolantonio Federica, Arena Sabrina, Gallicchio Margherita, Zecchin Davide, Martini Miriam, Flonta Simona Emilia, Stella Giulia Maria, Lamba Simona, Cancelliere Carlotta, Russo Mariangela, Geuna Massimo, Appendino Giovanni, Fantozzi Roberto, Medico Enzo, Bardelli Alberto
Laboratory of Molecular Genetics, Institute for Cancer Research and Treatment, University of Turin Medical School, I-10060 Candiolo, Turin, Italy.
Proc Natl Acad Sci U S A. 2008 Dec 30;105(52):20864-9. doi: 10.1073/pnas.0808757105. Epub 2008 Dec 23.
Mutations in oncogenes and tumor suppressor genes are responsible for tumorigenesis and represent favored therapeutic targets in oncology. We exploited homologous recombination to knock-in individual cancer mutations in the genome of nontransformed human cells. Sequential introduction of multiple mutations was also achieved, demonstrating the potential of this strategy to construct tumor progression models. Knock-in cells displayed allele-specific activation of signaling pathways and mutation-specific phenotypes different from those obtainable by ectopic oncogene expression. Profiling of a library of pharmacological agents on the mutated cells showed striking sensitivity or resistance phenotypes to pathway-targeted drugs, often matching those of tumor cells carrying equivalent cancer mutations. Thus, knock-in of single or multiple cancer alleles provides a pharmacogenomic platform for the rational design of targeted therapies.
癌基因和肿瘤抑制基因的突变是肿瘤发生的原因，也是肿瘤学中备受青睐的治疗靶点。我们利用同源重组将单个癌症突变敲入未转化的人类细胞基因组中。还实现了多个突变的顺序引入，证明了该策略构建肿瘤进展模型的潜力。敲入细胞表现出信号通路的等位基因特异性激活以及与通过异位癌基因表达获得的表型不同的突变特异性表型。对突变细胞上的一系列药理试剂进行分析，结果显示对靶向通路的药物有显著的敏感或耐药表型，通常与携带等效癌症突变的肿瘤细胞相匹配。因此，单个或多个癌症等位基因的敲入为靶向治疗的合理设计提供了一个药物基因组学平台。
