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

HT 筛选 T 细胞受体 CD3 界面的破坏

基本信息

批准号：
8507598
负责人：
Luc Teyton
金额：
$ 52.02万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8507598
关键词：
Address Adverse effects Affect Affinity Animal Experiments Animals Antibodies Antigen-Presenting Cells Autoimmunity Basic Science Binding Biochemistry Biological Biological Assay Biology Biophysics Blood Cells CD3 Antigens Cell Cycle Cell Surface Receptors Cell surface Cells Cellular Immunology Cellular biology Chemicals Chemistry Chronic Clinical Communicable Diseases Communication Complex Computer-Aided Design Cyclosporine Cysteine Development Disease Educational process of instructing Exhibits Extracellular Domain Family Fluorescence Polarization Funding Goals Healthcare Systems Human Immune response Immunity Immunologic Adjuvants Immunology Immunomodulators Immunosuppression Immunosuppressive Agents In Vitro Infection Injection of therapeutic agent Institutes Intravenous Knowledge Label Laboratories Lead Life Expectancy MHC Class I Genes MHC Class II Genes Maintenance Major Histocompatibility Complex Malignant Neoplasms Masks Medical Medicine Metabolic Pathway Minor Mixed Lymphocyte Culture Test Molecular Mus Neoplasms Nucleosides Organ Peptides Pharmaceutical Preparations Phase Process Property Quality of life Recombinants Regimen Series Side Signal Transduction Specialist Staging Steroids Surface System T-Cell Activation T-Cell Immunologic Specificity T-Cell Receptor T-Lymphocyte TCR Activation Technology Testing Therapeutic Toxic effect Translating Translational Research Translations Transplantation United States National Institutes of Health Work base combat cost dosage flexibility fluorophore graft vs host disease high throughput screening high throughput technology immunoregulation meetings mutant novel therapeutics programs receptor binding screening small molecule structural biology success synthetic drug tool

项目摘要

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.

描述（由申请人提供）：我们的项目通过应用高吞吐量技术来隔离能够破坏或修改愚蠢的T细胞受体（TCR）和CD3之间的小分子，直接响应程序目标。结构和生物物理研究已经告诉我们，TCR/CD3相互作用的亲和力非常低，对小改变非常敏感，这使其成为小分子发展的理想目标。我们使用单半半胱氨酸突变体上标记的重组TCR分子创建并优化了可及高吞吐量筛选的荧光偏振分析。概念验证活动隔离了6种化合物，这些化合物显示出具有TCR结合的特异性，并且能够特异性破坏TCR/CD3的T细胞通信，而不会影响T细胞信号传导。所有6个命中均抑制了MHC类I类和MHC II类限制的T细胞的T细胞信号传导，因此可以将其分类为免疫抑制化合物。我们的目标是确定化学探针，使我们能够进一步进一步了解T细胞激活的基本知识，并在同一过程中发现新的免疫调节剂家族。实际上，由于当前治疗的毒性，在移植和自身免疫的领域中急需新的免疫抑制药物。由于我们制定的策略，我们将分离的某些化学片段也可能表现出免疫刺激性能。这类药物，例如一般的免疫刺激剂尚不存在，并且会填补巨大的空白，以打击癌症和慢性传染病的免疫抑制。我们的项目说明了我们的研究所提供的一体化能力，通过将化学，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致力于将基础科学转化为新治疗学的开发。