Defining the relationship between kinase inhibitor pharmacophores and pro-/anti-targets using in vivo models of colorectal cancer

项目摘要 抗药性癌症是一个不幸的全球健康问题，影响着数百万人 家庭[6]。受体激酶域突变导致的靶向肿瘤抑制物 酪氨酸激酶通常以三磷酸腺苷结合位点为靶点[7]。虽然这种治疗方法是 通常非常有效，突变发生在“守门人”的位置，残基远离 药物结合部位不仅会导致药物结合亲和力减弱，而且还会增加 在没有药物的情况下的激酶活性。解决额外的激酶激活问题是一个主要问题 因为无法治愈的癌症很容易继续扩散。 我的论文研究旨在了解和量化这些突变所产生的影响 利用成纤维细胞中的激活域研究激活酶的活化和热力学稳定性 生长因子受体(FGFRs)家族作为模型系统。最近，我们阐明了一种变构 从铰链区到激活环的FGFR激酶残基网络， 都与激活有关。利用核磁共振弛豫实验提供特定的残基 信息，我计划量化横纹肌肉瘤中发现的守门人突变的影响， 和一个铰链突变体，最近发现严重削弱了抑制物结合，对激酶动力学。 这些动力学将为这些突变如何劫持激酶的变构提供一个清晰的图景 网络以提高活性并解释远离药物结合位点的铰链突变如何 进行抵抗。差示扫描量热法也将被用于了解药物如何 耐药突变会影响载脂蛋白和药物结合形式的激酶的稳定性。调查结果来自 这项工作将对未来的药物设计具有重大意义，以克服这些类型的药物 抗药性突变。

Project Summary Drug resistant cancers are an unfortunate global health problem that affects millions of families [6]. Inhibitors targeting cancers caused by mutations in the kinase domain of receptor tyrosine kinases often target the ATP binding site [7]. Although this method of treatment is generally very effective, mutations at the “gate-keeper” position and residues farther away from the drug binding site not only cause weakened binding affinity for the drug, but can also increase kinase activity in the absence of drug. Addressing additional kinase activation is a major concern as undruggable cancers can easily continue to spread. My dissertation research aims to understand and quantify the effects these mutations have on kinase activation and thermodynamic stability using the kinase domain from the fibroblast growth factor receptor (FGFRs) family as a model system. Recently, we elucidated an allosteric network of residues in FGFR kinases spanning from the hinge region to the activation loop that are associated with activation. Using NMR relaxation experiments to provide residue specific information, I plan to quantify the effects of the gate-keeper mutant found in rhabdomysarcoma, and a hinge mutant, recently found to severely weaken inhibitor binding, on kinase dynamics. These dynamics will provide a clear picture of how these mutations hijack the kinase's allosteric network to increase activity and explain how hinge mutations far from the drug-binding site can confer resistance. Differential scanning calorimetry will also be employed to understand how drug resistant mutations affect the stability of the kinase in the apo and drug-bound form. Findings from this work will have major implications for future drug design to overcome these types of drug resistance mutations.