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）和CD3（其必需的信号伙伴）之间通信的小分子，直接响应项目目标。结构和生物物理研究告诉我们，TCR/CD3相互作用具有非常低的亲和力，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适用于高通量筛选的荧光偏振测定方法，该方法使用重组TCR分子在单个半胱氨酸突变体上标记荧光团。一项概念验证活动分离出6种化合物，它们显示出TCR结合的特异性，并且能够特异性地破坏T细胞的TCR/CD3通讯，而不影响T细胞的信号传导。所有6个hit均抑制MHC I类和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，这将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新的免疫调节剂家族。事实上，由于目前治疗方法的毒性，移植和自身免疫领域急需新的免疫抑制药物。由于我们已经开发的策略，我们将分离的一些化学碎片也有可能表现出免疫刺激特性。这类药物，如一般免疫刺激剂，目前还不存在，将填补巨大的空白，以对抗癌症和慢性传染病的免疫抑制。我们的项目说明了我们研究所提供的整合能力，通过化学，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专家并肩工作，共同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.