Structural Studies of a T cell Specific Tyrosine Kinase

项目摘要 TEC家族激酶在造血来源的细胞中表达，并在造血干细胞中发挥关键作用。 在免疫细胞信号级联过程中的发展和免疫激活。两个TEC 激酶，ITK和BTK是本申请的焦点，并且都被认为是 治疗免疫相关疾病的重要目标。例如，活动站点 BTK抑制剂，伊曲替尼，用于治疗慢性淋巴细胞白血病，但新的方法 因为耐药性正在挫败减缓疾病进展的努力。当前 续期申请采取多方面的方法处理这个问题。首先我们 利用上一个奖励期的机械发现， 用于小分子发现的几个正交筛选。到目前为止的发现表明， 已经成功地鉴定了与TEC激酶结合的小分子试剂， 调节T和B细胞信号传导。筛选方法旨在揭示 激酶活性的变构调节剂，并以这种方式，我们的目标是开发的手段， 用联合疗法克服伊鲁替尼耐药性，以及通常定义 替代的-活性位点之外-激酶抑制的方法。的额外 研究目标，我们使用我们的生物物理工具库（NMR光谱， 氢/氘交换质谱法和X射线晶体学）。 生物化学和细胞分析，以了解如何驱动抗药性的突变， 或致癌性改变全长ITK和BTK的结构、动力学和功能 激酶。从这项工作中产生的分子水平的知识将提供一个 更好地理解T细胞和B细胞信号传导以及靶向特异性 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.