Project 3: Anti-Inflammatory Mechanisms of Inhaled Hypertonic Saline

项目3：吸入高渗盐水的抗炎机制

基本信息

批准号：
8382283
负责人：
ANIRBAN BANERJEE
金额：
$ 36.63万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-06-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8382283
关键词：
Acute Adhesions Adhesives Affect Aftercare Alveolar Alveolar Cell Animals Anti-Inflammatory Agents Anti-inflammatory Asses Attenuated Automobile Driving Bioinformatics Blood Breathing CCAAT-Enhancer-Binding Proteins Cell Nucleus Cell model Cells Clathrin Complex Cystic Fibrosis Data Databases Development Exons Family Family member Fluorescence Resonance Energy Transfer Gene Expression Genes Genetic Transcription Genome Goals Hemorrhage Human IRF1 gene Immunoprecipitation Importins Inflammatory Infusion procedures Interleukin-1 Kinetics Leukocytes Libraries Lung Lung Inflammation NF-kappa B Nuclear Import Nuclear Translocation Patients Peptides Phenotype Principal Investigator Production Proteins Proteomics RNA Interference Receptor Signaling Regulation Reporter Resuscitation Reverse Transcriptase Polymerase Chain Reaction Safety Saline Scheme Shock Signal Transduction Site Stimulus Testing Therapeutic Time Translating Trauma cell type chemokine cytokine density dimer dosage lung injury macrophage member molecular phenotype neutrophil novel p65 pneumocyte prevent programs promoter receptor research study trafficking transcription factor

项目摘要

Trauma, hemorrhage and blood resuscitation produce acute lung inflammation by inducing chemokine, cytokine and, and adhesive proteins. Up-regulating mechanisms depend on systemic signals that translocate gene-regulating, transcription factors (TFs), such as NF-kB into the nucleus. The genes expressed direct the adhesion, transmigration of leukocytes and phenotypic developments in the alveolar milieu. Hyperosmolarity (HOsm), appears to prevent inflammatory gene expression but the mechanisms and scope are unclear. Therapeutic hypertonic saline infusions during trauma-resuscitation, or inhaled as for cystic fibrosis, are well tolerated. We hypothesize that hyperosmolarity alters the nuclear translocation of selected transcription factors induced by inflammatory stimuli by promoting novel complexes. In this proposal period (2005-2010) we will first examine the traffic of TFs controlling prototypical alveolar cytokines and adhesive proteins that are modified by HOsm, and investigate complexes of these TFs, including those formed with importins. 1. Investigate HOsm altered translocation of the NF-KB and IKK family members, their HOsm altered cytoplasmic complexes and examine alternate complexes promoted by HOsm with IP-MS proteomics. 2. Since HOsm does not prohibit p65 Rel A translocation induced by IL-1 yet suppresses certain cytokines, we will examine translocation of other essential promoters of the IRF, C/EBP. and R/FLAT families and identify their HOsm altered partners. Next, to expand the inquiry beyond the known cytokines we will execute: 3. Gene array analyses, to guide bioinformatics queries for potential TFs. The goal is to gradually build up a library of TFs that do not translocate successfully during HOSm and identify their sequestering partners, (starting with NF-kB and IRF family members). Lastly, we will translate these bench experiments to asses the tolerability or benefit of nebulized HOsm in shocked animals and trauma patients. 4. Evaluate lung inflammation and TF translocation in traumatized Animal or Patients treated with inhaled HOsm. The information from Aims 1-3 will be used to interpret the molecular phenotypes and TF redistributions after treatments.

创伤，出血和血液复苏通过诱导趋化因子，细胞因子和粘附蛋白而产生急性肺炎症。上调机制取决于易位的基因调节，转录因子（TFS）的系统信号，例如NF-KB进入核。这些基因表达了肺泡环境中白细胞和表型发展的粘附，迁移。高渗透性（HOSM）似乎可以防止炎症基因表达，但机制和范围尚不清楚。良好的耐受性，可以耐受性抗创伤时的治疗性高渗盐水输注或吸入囊性纤维化。我们假设高渗透性通过促进新型复合物来改变由炎症刺激引起的选定转录因子的核转运。在此提案期（2005-2010）中，我们将首先研究由HOSM修饰的原型肺泡细胞因子和粘附蛋白的TFS流量，并研究这些TFS的复合物，包括用importin形成的TF。 1。研究HOSM改变了NF-KB和IKK家族成员的易位，其HOSM改变了细胞质复合物，并检查了由IP-MS蛋白质组学促进的替代复合物。 2。由于HOSM不禁止p65与IL-1诱导但抑制某些细胞因子的易位，因此我们将检查IRF，C/EBP的其他必要启动子的易位。和R/Flat家庭，并确定其HOSM改变的合作伙伴。接下来，为了将查询扩展到已知的细胞因子之外，我们将执行： 3。基因阵列分析，以指导潜在TF的生物信息学查询。目的是逐渐建立一个在HOSM期间无法成功转移并确定其隔离伙伴的TF库（从NF-KB和IRF家庭成员开始）。最后，我们将翻译这些台式实验，以评估震惊的动物和创伤患者中雾化的HOSM的耐受性或益处。 4。评估受吸入HOSM治疗的创伤动物或患者的肺部炎症和TF易位。 AIMS 1-3的信息将用于解释治疗后的分子表型和TF重新分布。