CD47 Regulation of Leukocyte Integrins during Leukocyte Trafficking

白细胞贩运过程中 CD47 对白细胞整合素的调节

基本信息

批准号：
9105867
负责人：
FRANCIS W. LUSCINSKAS
金额：
$ 58.46万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-01 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9105867
关键词：
Address Adhesions Adhesives Affect Affinity Alanine Anti-Inflammatory Agents Anti-inflammatory Antibodies Antigen-Presenting Cells Antigens Atherosclerosis Autoimmune Diseases Binding Biochemical Biological Biological Assay Blood Platelets C-terminal CD4 Positive T Lymphocytes CD47 gene Cardiovascular Diseases Cell Culture System Cell physiology Cell surface Cells Chronic Clinical Trials Colitis Collaborations Complex Data Defect Development Diabetes Mellitus Disease Effector Cell Employee Strikes Endothelium Epitopes Escherichia coli Experimental Autoimmune Encephalomyelitis Extracellular Matrix Extracellular Matrix Proteins Generations Glycoproteins Goals Graft Rejection Haptens Homeostasis Homing Host Defense Human Human Cell Line Hypertension Immigration Immune In Vitro Inflammation Inflammatory Inflammatory Response Injury Integrin alpha4beta1 Integrin alphaVbeta3 Integrins Intercellular adhesion molecule 1 Ischemia Jurkat Cells Kidney Leukocyte Trafficking Leukocytes Ligands Lung Inflammation Lymphocyte Lymphoid Tissue Malignant Neoplasms Mass Spectrum Analysis Measures Mediating Mediator of activation protein Metabolic syndrome Modeling Molecular Molecular Conformation Mus Mutagenesis Neoplasm Metastasis Peptide Fragments Peptides Phage Display Pharmaceutical Preparations Phenotype Physiological Placenta Play Process Proteins Publications Reaction Reagent Regulation Reperfusion Therapy Reporting Research Personnel Role SHPS-1 protein Sampling Scanning Signal Pathway Signal Transduction Signaling Molecule Site Skin graft Stroke Surface T cell regulation T-Lymphocyte T-Lymphocyte Subsets TNF gene Technology Th1 Cells Therapeutic Thick Thrombospondin 1 Tissues Vascular Cell Adhesion Molecule-1 base cell type chemokine cremaster muscle cytokine design in vitro Model in vivo innovation insight migration novel public health relevance therapeutic target trafficking

项目摘要

DESCRIPTION (provided by applicant): Inflammation is a significant contributor to cardiovascular disease and related pathophysiological processes including stroke, hypertension, atherosclerosis, diabetes-metabolic syndrome. The activation and recruitment of effector T cell subsets into tissues is a highly regulated process that is dependent upon adhesive interactions between T cell subsets and the endothelium lining the vascularture. Therefore, our overall goal is to gain a better understanding of the mechanisms that regulate CD4+ T cell subset recruitment to sites of immune-mediated inflammation. Based on our exciting preliminary data and recent publication, we hypothesize that CD47 plays an important role in leukocyte β1 and β2 integrin adhesive functions by regulating the expression of high affinity conformations. By virtue of its effect on LFA-1 and VLA-4 integrin adhesive function and its well-documented role in immune cell homeostasis, CD47 and its ligand SIRPα play a critical role in multiple steps (arrest and migration) in the inflammatory response. The specific aims will determine the molecular mechanisms and underlying structural basis of CD47 regulation of T cell LFA-1 and VLA-4 adhesive function and its role in T effector cell functions. We propose complementary and well-characterized in vitro cell culture systems and novel CyTOF mass spectroscopy technology to dissect the biochemical and molecular signaling pathways used by CD47 in human and murine immune cells to regulate integrin function. Specifically, Specific Aim 1 will employ recently developed single cell mass spectrometry (CyTOF mass spectrometry) and standard biochemical assays to interrogate intracellular signaling pathways in CD47-/- T cells to identify defects that cause impaired integrin activation in CD47-/- leukocytes. Up to 40 cell-surface molecules, intracellular signaling molecules and mediators (cytokines) can be measured in a single sample using CyTOF to provide much more information about cell phenotypes at a single-cell level. Specific Aim 2 will address whether in trans CD47 ligands, Signal Regulatory Protein (SIRP)α, and SIRPγ, and the secreted extracellular matrix protein, thrombospondin-1, affect CD47 in cis regulation of LFA-1 and VLA4 function during T cell recruitment. In Specific Aim 3 we propose to determine the residues in CD47 that interact with β2 integrins, which may provide guidance for designing novel and innovative therapeutic target(s), and identify biological reagents that impair these interactions. These approaches and our collaborations with established investigators will generate novel insights into CD47 regulation of integrin activation and adhesive functions in T cell trafficking.

描述（由适用提供）：炎症是心血管疾病和相关病理生理过程的重要原因，包括中风，高血压，动脉粥样硬化，糖尿病 - 代谢综合征。效应T细胞亚群进入组织的激活和募集是一个高度调节的过程，取决于T细胞亚群和脉管体衬里的粘合性相互作用。因此，我们的总体目标是更好地了解将CD4+ T细胞子集募集到免疫介导的感染部位的机制。根据我们令人兴奋的初步数据和最近的发表，我们假设CD47通过调节高亲和力构象的表达来表达白细胞β1和β2整合素粘合剂功能在白细胞β1和β2整合蛋白粘附功能中起重要作用。由于它对LFA-1和VLA-4整联蛋白粘附功能的影响以及其在免疫细胞稳态中有据可查的作用，CD47及其配体SIRPα在炎症反应中的多个步骤（逮捕和迁移）中起着至关重要的作用。具体目的将确定CD47调节T细胞LFA-1和VLA-4粘附功能的分子机制和基础结构基础及其在T效应细胞功能中的作用。我们提出了完整且良好的体外细胞培养系统和新型的质谱质谱技术，以剖析CD47在人和鼠类免疫细胞中用于调节整联蛋白功能的生化和分子信号通路。具体的，特定的目标1将采用最近开发的单细胞质谱法（质谱质谱）和标准的生化测定法来询问CD47 - / - T细胞中细胞内信号传导途径，以识别导致CD47 - / - 白细胞中整联蛋白激活受损的缺陷。可以在单个样品中使用Cytof在单个样品中测量多达40个细胞表面分子，细胞内信号分子和介体（细胞因子），以在单细胞水平上提供有关细胞表型的更多信息。具体的目标2将解决反式CD47配体，信号调节蛋白（SIRP）α和SIRPγ以及分泌的细胞外基质蛋白血小板蛋白1是否会影响CIS在T细胞募集过程中的LFA-1和VLA4功能中CD47。我们建议的特定目的3我们建议确定与β2整合素相互作用的CD47中的残差，这可能为设计新颖和创新的治疗靶靶标提供指导，并确定损害这些相互作用的生物试剂。这些方法以及我们与既定研究者的合作将产生对T细胞运输中整合素激活和粘合功能的CD47调节的新见解。