Structural Studies of a T cell Specific Tyrosine Kinase

T 细胞特异性酪氨酸激酶的结构研究

基本信息

批准号：
10459324
负责人：
AMY H ANDREOTTI
金额：
$ 45.2万
依托单位：
IOWA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-01-15 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10459324
关键词：
Active Sites Address Agammaglobulinaemia tyrosine kinase Allosteric Site Antigen Receptors Autoimmunity Award B-Cell Antigen Receptor B-Lymphocytes Binding Biochemical Catalytic Domain Cell Line Cell physiology Cells Cellular Assay Chemicals Chronic Lymphocytic Leukemia Combined Modality Therapy Communication Cues Data Deletion Mutation Deuterium Development Disease Disease Progression Drug resistance FDA approved Family Gene Expression Goals Grant Hematopoietic Hematopoietic Neoplasms Hydrogen Immune Immune System Diseases Immune response Immunosuppression Inflammatory Inflammatory Response Interleukin-2 Iowa Knowledge Lead Length Life Mantle Cell Lymphoma Mass Spectrum Analysis Massachusetts Mediating Methodology Modification Molecular Molecular Conformation Motion Mutation NMR Spectroscopy Nature Oncogenic Organic Chemistry Pathogenicity Patients Pharmaceutical Preparations Phosphorylation Phosphotransferases Play Protein Isoforms Protein Tyrosine Kinase Proteins Publishing Reagent Receptor Signaling Regulation Reporting Research Resistance Resistance development Role Signal Transduction Signaling Protein Site Structure Synthesis Chemistry T-Cell Receptor T-Lymphocyte TEC Protein Tyrosine Kinase Therapeutic Intervention Therapeutic Uses Translating Tyrosine Kinase Inhibitor Tyrosine Phosphorylation Universities Variant Waldenstrom Macroglobulinemia Wales Work X-Ray Crystallography base biophysical tools chronic graft versus host disease design immune activation inhibitor intermolecular interaction kinase inhibitor novel strategies programs protein complex public health relevance resistance mechanism response screening small molecule targeted treatment tool

项目摘要

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.

项目概要 TEC 家族激酶在造血来源细胞中表达并发挥关键作用发育和免疫激活过程中免疫细胞信号级联反应中的两个 TEC。激酶、ITK 和 BTK 是本应用的重点，并且都被认为是治疗免疫相关疾病的重要目标，例如活性位点。 BTK抑制剂依鲁替尼用于治疗慢性淋巴细胞白血病但新方法由于耐药性正在阻碍减缓疾病进展的努力，因此需要采取这些措施。续订申请采取多方面的方法来解决这个问题。利用上一个奖励期的机械结果，执行迄今为止的发现表明我们对小分子发现进行了一些正交筛选。已成功鉴定出与 TEC 激酶结合的小分子试剂，筛选方法旨在揭示调节 T 细胞和 B 细胞信号传导。激酶活性的变构调节剂，通过这种方式，我们的目标是开发方法通过联合疗法克服依鲁替尼耐药性以及一般定义另一种方法是在活性位点之外进行激酶抑制。为了实现研究目标，我们使用我们的生物物理工具库（核磁共振波谱、另外，氢/氘交换质谱法和 X 射线晶体学）通过生化和细胞分析来了解突变如何驱动耐药性或致癌性改变全长 ITK 和 BTK 的结构、动力学和功能这项工作中产生的分子水平知识将提供更好地了解 T 细胞和 B 细胞信号传导以及针对特定目标的方法用于治疗用途的相互作用或变构调节机制。