HT screens for the disruption of the T cell receptor CD3 interface

描述（由申请人提供）：我们的项目直接响应计划目标，通过应用高通量技术分离能够破坏或修饰独特型T细胞受体（TCR）与其专性信号伴侣CD 3之间通讯的小分子。结构和生物物理学研究告诉我们，TCR/CD 3相互作用的亲和力非常低，对微小的改变非常敏感，使其成为小分子开发的完美靶点。我们已经创建并优化了一种荧光偏振测定法，该测定法适用于使用在单个半胱氨酸突变体上用荧光团标记的重组TCR分子进行高通量筛选。概念验证活动分离出6种化合物，其显示出对TCR结合的特异性，并且能够特异性破坏T细胞的TCR/CD 3通讯而不影响T细胞信号传导。所有6种命中物均抑制MHC I类限制性T细胞和MHC II类限制性T细胞的T细胞信号传导，因此可归类为免疫抑制化合物。我们的目标是确定化学探针，使我们能够进一步了解T细胞活化的基本知识，并在同一过程中发现免疫调节剂的新家族。事实上，由于目前治疗的毒性，在移植和自身免疫领域非常需要新的免疫抑制药物。由于我们已经开发的策略，我们将分离的一些化学片段也可能表现出免疫刺激特性。这类药物，例如一般的免疫刺激剂，还不存在，将填补一个巨大的空白，以打击癌症和慢性传染病的免疫抑制。我们的项目展示了我们研究所提供的整合能力，该研究所拥有化学，结构生物学，生物物理学，化学生物学，细胞免疫学和动物免疫学方面的并排专家，共同努力实现将基础科学转化为新疗法开发的明确目标。

DESCRIPTION (provided by applicant): Our project is directly responsive to the program objectives by applying high throughput technologies to isolate small molecules capable of disrupting or modifying the communication between the idiotypic T cell receptor (TCR) and CD3, its obligate signaling partner. Structural and biophysical studies have taught us that the TCR/CD3 interaction was of very low affinity and very sensitive to minor alterations, making it a perfect target for small molecules development. We have created and optimized a fluorescence polarization assay amenable to high throughput screen using recombinant TCR molecules labeled with fluorophore on single cysteine mutants. A proof of concept campaign isolated 6 compounds that showed specificity for TCR binding and were capable of disrupting specifically the TCR/CD3 communication of T cells without affecting T cells signaling. All 6 hits were inhibiting T cell signaling of both MHC class I- and MHC class II-restricted T cells and as such could be categorized as immunosuppressive compounds. Our goals are to identify chemical probes that will allow us to further our basic knowledge of T cell activation and to discover, in that same process, new families of immunomodulators. Indeed, new immunosuppressive drugs are much needed in the field of transplantation and autoimmunity, because of the toxicity of the current treatments. Because of the strategy that we have developed, it is also possible that some chemical fragments that we will isolate, will exhibit immunostimulatory properties. This class of drugs, e.g. general immunostimulants, does not exist yet and would fill a huge gap to combat the immunosuppression of cancer and chronic infectious diseases. Our project illustrates the capabilities of integration that our institute offers by having side-by-side specialists in chemistry, structural biology, biophysics, chemical biology, cellular immunology and animal immunology working together towards a clearly defined goal of translating basic science into the development of new therapeutics.