Molecular and Cellular Targets of Adenosine in Lung

肺中腺苷的分子和细胞靶标

基本信息

批准号：
7415116
负责人：
Joel M. Linden
金额：
$ 32.97万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-05-01 至 2009-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7415116
关键词：
ADORA2A gene ADORA3 gene ATL 146e Acute Adenosine Adenosine A2A Receptor Adherence Adhesions Adoptive Transfer Adrenal Cortex Hormones Affect Agonist Alkylating Agents Alveolar Macrophages Animals Anti-Inflammatory Agents Anti-inflammatory Antibiotic Therapy Antigens Binding Biological Assay Bleomycin Blood Blood Pressure Blood Vessels Bolus Infusion Bone Marrow Bone Marrow Transplantation CCL3 gene CCL4 gene CD8B1 gene CXCL10 gene CXCL2 gene Cell Adhesion Molecules Cell Communication Cell Count Cell surface Cells Cellular Assay Cessation of life Characteristics Chimera organism Collagen Companions Complex Custom Cyclic AMP DNA Data Detection Dose Drug or chemical Tissue Distribution Edema Endothelial Cells Escherichia coli Event Experimental Designs Exposure to Fibrosis Gene Chips Generations Genes Genetic Genotype Goals Heart Heart Rate Hepatic Heterotrimeric G Protein Subunit Heterotrimeric GTP-Binding Proteins Histologic Hour Human IL8RB gene Image Immunohistochemistry Immunosuppressive Agents In Situ Hybridization Inbred BALB C Mice Individual Infectious Agent Inflammation Inflammatory Inflammatory Response Infusion Pumps Injection of therapeutic agent Injury Injury to Kidney Integrins Interleukin-1 Interleukin-2 Interleukin-4 Interleukin-5 Intraperitoneal Injections Knock-out Knockout Mice L-Selectin Learning Letters Leukocyte Trafficking Leukocytes Life Ligands Liver Lung Lung Inflammation Magnetic Resonance Imaging Malignant Neoplasms Marrow Measures Mediating Messenger RNA Modeling Molecular Monitor Mouse Strains Mus NOS2A gene Peritoneal Macrophages Phenotype Physiological Pneumonia Polymerase Chain Reaction Population Principal Investigator Procedures Production Purinergic P1 Receptors Purpose RANTES RNA Rain Reactive Oxygen Species Receptor Gene Regulation Reperfusion Injury Reporting Resistance Respiratory Burst Reverse Transcriptase Polymerase Chain Reaction Role Role playing therapy Sampling Score Sepsis Skin Small Inducible Cytokine A3 Smooth Muscle Myocytes Sorting - Cell Movement Source Spinal Cord Spleen Steroids Stimulus Surface T-Lymphocyte Therapeutic Thinking Tilia Time Tissues Toxic effect Toxin Transcript Transcriptional Activation Universities Up-Regulation Vascular Endothelial Cell Vascular Permeabilities Vasodilator Agents Wild Type Mouse adenosine receptor activation aerosolized cell type cellular targeting chemokine chemokine receptor cytokine free radical oxygen genetically modified cells granulocyte heart function human NOS2A protein ilium improved in vivo indium-bleomycin inhibitor/antagonist interest knock-down lung injury macrophage mouse model neutrophil osteopontin programs protective effect radioligand receptor receptor binding receptor expression research study response tool trafficking ultra high resolution

项目摘要

"Molecular and Cellular Targets of Adenosine in Lung" will focus on identifying inflammatory cells that are important for mediating bleomycin toxicity to the lung, and the mechanisms used by agonists of the A2A adenosine receptor (A2AAR) to inhibit lung inflammation. Aim 1A is to characterize cells from genetically modified mice with tissue specific deletions of the A2AAR in bone marrow-derived cells, T cells, granulocytes or endothelial cells (EC). Transcripts for the four adenosine receptor (AR) subtypes will be measured by quantitative RT-PCR in purified neutrophils, macrophages, T cells and lung EC purified. Aim 1B is to phenotype A2AAR responses in cells derived from these mice is functional assays of neutrophils (oxidative burst), macrophages (TNFalpha release and integrin expression), T cells (INFgamma release and adherence) and EC (adherence and adhesion molecule expression) or FACS detection of cell surface activation markers. Hypothesis 1 is that it will be possible to nearly eliminate A2AAR expression and function in cells derived from various genetically modified mice with little effect on the expression other AR subtypes. These studies will be helped by the availability of unique AR subtype selective ligands. Aim 2 is to examine the induction in response to LPS (positive control) and bleomycin of mRNAs induced by inflammation in macrophages and other cells. Induced ARs will be quantified by mRNAs, radioligand binding (where possible), and functional assays. Hypothesis 2 is that functional anti-inflammatory responses to adenosine are strongly induced by inflammatory stimuli. Aim 3 is to determine how bleomycin and A2AAR activation affect leukocyte trafficking, pulmonary cytokine production and fibrosis in vivo using mice with various targeted A2AAR deletions, and mice lacking pulmonary macrophages, INFgamma or CXCR2 receptors (that bind KC or MIP-1alpha). Cell trafficking will be measured by counting cells is BALF, dispersed lung, immunohistochemistry and, in live animals by ultra-high resolution gamma-imaging and MRI. Hypothesis 3 is that the response to A2AAR activation influenced by pulmonary macrophages and/or T cells. These results will be helpful for determining the role of A2AAR activation in protecting lungs from other injuries (LPS and ischemia-reperfusion injury) that are being investigated in the other projects.

“肺中腺苷的分子和细胞靶标”将重点鉴定对于介导博来霉素肺部毒性很重要的炎症细胞，以及 A2A 腺苷受体 (A2AAR) 激动剂抑制肺部炎症的机制。目标 1A 是鉴定来自骨髓来源细胞、T 细胞、粒细胞或内皮细胞 (EC) 中 A2AAR 组织特异性缺失的转基因小鼠的细胞。将通过定量 RT-PCR 在纯化的中性粒细胞、巨噬细胞、T 细胞和纯化的肺 EC 中测量四种腺苷受体 (AR) 亚型的转录本。目标 1B 是对源自这些小鼠的细胞中的 A2AAR 反应进行表型分析，对中性粒细胞（氧化爆发）、巨噬细胞（TNFα 释放和整联蛋白表达）、T 细胞（INFγ 释放和粘附）和 EC（粘附和粘附分子表达）进行功能分析，或FACS检测细胞表面活化标记物。假设 1 是有可能几乎消除来自各种转基因小鼠的细胞中的 A2AAR 表达和功能，而对其他 AR 亚型的表达几乎没有影响。独特的 AR 亚型选择性配体的可用性将有助于这些研究。目标 2 是检查巨噬细胞和其他细胞中炎症诱导的 mRNA 对 LPS（阳性对照）和博来霉素的诱导反应。诱导的 AR 将通过 mRNA、放射性配体结合（如果可能）和功能来量化化验。假设2是腺苷的功能性抗炎反应是由炎症刺激强烈诱导的。目标 3 是使用具有各种靶向 A2AAR 缺失的小鼠以及缺乏肺巨噬细胞、INFgamma 或 CXCR2 受体（结合 KC 或 MIP-1alpha）的小鼠，确定博来霉素和 A2AAR 激活如何影响体内白细胞运输、肺细胞因子产生和纤维化。细胞运输将通过 BALF、分散肺、免疫组织化学以及活体动物中的超高分辨率伽马成像和 MRI 细胞计数来测量。假设 3 是对 A2AAR 激活的反应受到肺巨噬细胞和/或 T 细胞的影响。这些结果将有助于确定 A2AAR 激活在保护肺部免受其他损伤（LPS 和缺血再灌注损伤）中的作用，其他项目正在研究这些损伤。