蛋白激酶介导的底物磷酸化是重要的翻译后修饰之一，参与调控几乎所有的生物学过程和通路。蛋白激酶是癌症治疗的重要药物靶标，其活性异常与肿瘤的发生、发展和转移密切相关。激酶活性由酶反应活性和底物识别特异性两部分构成，其相关研究即癌症激酶组活性分析已成为重要挑战，有望为癌症的个性化医疗提供潜在的、更有效的药靶和治疗方案。本项目中，我们拟结合非标记定量磷酸化蛋白质组鉴定技术、生物信息学预测方法以及分子、生化、细胞和小鼠实验等研究手段，分别鉴定和定量多种人类正常肝细胞和肝癌细胞系在索拉非尼处理前后的磷酸化位点及修饰水平，设计新的肝癌激酶组活性分析的计算方法，建立整合的计算分析与实验验证平台，准确预测并证实肝癌中活性显著差异的激酶，发现参与新的、调控肝癌转移和耐药性的激酶，利用相应的小分子化合物探索单独或联合用药的潜在治疗方案，并在细胞和小鼠中验证其对肝癌抑制的有效性。

Substrate phosphorylation mediated by protein kinases is one of the most important post-translational modifications, and involved in regulating almost all of biological processes and pathways. Protein kinases have been identified as key drug targets for cancer therapy, and the abnormality of kinase activity is highly associated with tumorigenesis, cancer progression and metastasis. The protein kinase activity is composed by enzymatic activity and the specificity of substrate recognition, whereas related researches such as kinome activity profiling in cancers have been emerged to be great challenges, and can provide potentially more efficient drug targets and treatments for personalized medicine of cancers. In this project, we will combine multiple techniques, such as non-label quantitative phosphoproteomic identification, bioinformatic prediction, and various experimental approaches of molecular biology, biochemistry, cell biology and mouse model, to characterize and quantify phosphorylation sites with modification levels in a variety of human normal liver and hepatocellular carcinoma (HCC) cell lines. We will design novel computational algorithms for profiling kinome activity in liver cancers, develop an integrative platform for both computation and experiments, and accurately predict significantly activated or inhibited kinases in HCCs, together with following experimental validations. Based on the results, we will experimentally identify new protein kinases that participate in the regulation of HCC metastasis and drug-resistance. Using small-molecule compounds of newly identified kinases, we will design potentially useful treatments by individual or combinatorial drugs, and probe the efficacy on HCC inhibition in both cell and mouse levels.