Structural Studies of a T cell Specific Tyrosine Kinase

项目摘要 TEC家族激酶在造血细胞中表达，并发挥关键作用 在免疫细胞中，信号在发育和免疫激活过程中级联。两个TEC 激酶、ITK和BTK是本申请的重点，它们都被认为是 在治疗免疫相关疾病的努力中的重要目标。例如，活动站点 BTK抑制剂伊布鲁替尼用于治疗慢性淋巴细胞白血病 是必要的，因为抗药性正在挫败减缓疾病进展的努力。海流 续签申请采取了多方面的方法来解决这个问题。首先，我们是 利用上一个获奖期的机械调查结果，通过执行 用于小分子发现的几个正交屏幕。到目前为止的发现表明，我们 已成功鉴定出与TEC激酶和TEC蛋白结合的小分子试剂 调节T细胞和B细胞信号。筛选方法旨在揭示 变构调节剂的激酶活性，通过这种方式，我们的目标是开发手段来 通过联合治疗克服伊布鲁替尼耐药性，以及一般定义 活性部位以外的替代方法抑制激酶。在附加内容中 研究目标，我们使用我们的生物物理工具(核磁共振波谱， 氢/氢交换质谱仪和x射线结晶学)此外 到生化和细胞分析，以了解导致耐药性的突变 或致瘤性改变全长ITK和BTK的结构、动力学和功能 激活剂。从这项工作中产生的分子水平的知识将提供一个 更好地了解T细胞和B细胞信号以及靶向特定信号的方法 用于治疗用途的相互作用或变构调节机制。

Project Summary The TEC family kinases are expressed in cells of hematopoietic origin and play key roles in immune cell signaling cascades during development and immune activation. Two TEC kinases, ITK and BTK are the focus of this application and are both recognized as important targets in efforts to treat immune related diseases. For example, the active site BTK inhibitor, ibrutinib, is used to treat chronic lymphocytic leukemia but new approaches are needed as drug resistance is foiling efforts to slow progression of disease. The current renewal application takes a multifaceted approach to this problem. First we are capitalizing on the mechanistic findings from the previous award period by carrying out several orthogonal screens for small molecule discovery. The findings so far suggest we have successfully identified small molecule reagents that bind to the TEC kinases and modulate T- and B-cell signaling. The screening approaches are designed to uncover allosteric modulators of kinase activity and in this way we aim to develop the means to overcome ibrutinib resistance with combination therapies as well as generally define alternative – outside of the active site - approaches to kinase inhibition. In additional research objectives, we use our arsenal of biophysical tools (NMR spectroscopy, hydrogen/deuterium exchange mass spectrometry, and x-ray crystallography) in addition to biochemical and cellular assays to understand how mutations that drive drug resistence or oncogenicity alter the structure, dynamics and function of the full-length ITK and BTK kinases. The molecular level knowledge that will emerge from this work will provide a better understanding of T cell and B cell signaling and the means to target specific interactions or allosteric regulatory mechanisms for therapeutic uses.