DDR2 kinase inhibition in squamous cell lung carcinomas

项目总结（见说明）： 在我们的初步研究中，我们发现DDR2，盘状结构域受体2酪氨酸激酶基因，在3%至4%的肺鳞状细胞癌（肺SCC）中发生体细胞突变。DDR2的敲低杀死携带DDR2突变的肺SCC细胞。此外，用本发明的化合物处理DDR2突变型肺SCC细胞， 酪氨酸激酶抑制剂达沙替尼显著降低细胞增殖;通过携带达沙替尼抗性T654 M看门人突变的DDR2的过表达来挽救达沙替尼的作用。 我们现在的目标是进一步开发DDR2作为肺SCC的治疗靶点。我们的具体目标是与其他项目和共享资源核心合作，如下所示： - 具体目标1.我们将使用细胞和动物模型系统来表征肿瘤衍生的DDR2突变体在肺基底上皮中的致癌潜力。我们将与Core C（动物）合作，以产生肺基底上皮特异性表达DDR2突变体的转基因小鼠模型，并测定肿瘤形成。我们还将在基底肺上皮中表达DDR2突变体mRNA，并检测致癌作用。这些动物和细胞模型将用于评估通过遗传修饰和小分子抑制剂破坏DDR2功能的影响。 - 具体目标2.我们将在肺鳞癌模型中评估DDR2抑制剂达沙替尼耐药的机制。为了研究对达沙替尼的c/s耐药，我们将在DDR2中设计和/或选择达沙替尼耐药等位基因，并测试它们对达沙替尼和其他激酶抑制剂的敏感性。此外，我们将通过开放阅读框架文库的异位表达来筛选反式抗性。 - 具体目标3.使用生物化学和细胞分析开发DDR2的有效和选择性抑制剂，其效果。我们将与核心A（化学）和核心B（结构）合作，优化核心A鉴定的先导化合物，作为MDR 2的MUR特异性抑制剂，相对于达沙替尼，具有更高的DDR2激酶特异性。这些优化的抑制剂将在肺SCC衍生的细胞系以及Aim 1中开发的细胞和动物模型系统中进行测试，包括对达沙替尼耐药的工程模型。 通过这些努力，该项目将推进DDR2抑制剂作为肺鳞状细胞癌潜在治疗方法的开发。

PROJECT SUMMARY (See instructions): In our Preliminary Studies, we found that DDR2, the discoidin domain receptor 2 tyrosine kinase gene, is somatically mutated in 3 to 4% of lung squamous cell carcinomas (lung SCC). Knock-down of DDR2 kills lung SCC cells bearing DDR2 mutations. Furthermore, treatment of DDR2-mutant lung SCC cells with the tyrosine kinase inhibitor dasatinib significantiy decreases cell proliferation; the effects of dasatinib are rescued by over-expression of DDR2 bearing the dasatinib-resistant T654M gatekeeper mutation. We now aim to develop DDR2 further as a therapeutic target for lung SCC. Our specific aims are to work with the other Projects and the shared resource Cores as follows: -Specific Aim 1. We will characterize the oncogenic potential of tumor-derived DDR2 mutants in the basal epithelium of the lung using cellular and animal model systems. We will work with Core C (Animal) to generate transgenic mouse models for lung basal epithelium-specific expression of DDR2 mutants and assay for tumor formation. We will also express DDR2 mutant mRNA in basal lung epithelium and test for oncogenic effects. These animal and cellular models will be used to assess the impact of disruption of DDR2 function both by genetic modification and by small molecule inhibitors. -Specific Aim 2. We will evaluate mechanisms of resistance to the DDR2 inhibitor, dasatinib, in squamous lung cancer models. To study c/s resistance to dasatinib, we will engineer and/or select for dasatinib resistance alleles in DDR2 and test their sensitivities to dasatinib and other kinase inhibitors. In addition, we will screen for trans resistance by ectopic expression of open reading frame libraries. -Specific Aim 3. Develop potent and selective inhibitors of DDR2 using biochemical and cellular assays, their effects. We will collaborate with Core A (Chemistry) and Core B (Structure) to optimize lead compounds identified by Core A as mutant-specific inhibitors of DDR2 with greater DDR2 kinase specificity relative to dasatinib. These optimized inhibitors will be tested in both lung SCC-derived cell lines and in the cellular and animal model systems developed in Aim 1, including models engineered to be resistant to dasatinib. Through these efforts, this project will advance the development of DDR2 inhibitors as potential treatments for squamous cell carcinomas of the lung.