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产生急性肺部炎症。上调机制依赖于将基因调节转录因子 (TF)（例如 NF-kB）转入细胞核的系统信号。这些基因表达直接白细胞的粘附、迁移和肺泡环境中的表型发育。高渗透压（HOsm）似乎可以阻止炎症基因表达，但其机制和范围尚不清楚。创伤复苏期间的治疗性高渗盐水输注或吸入治疗囊性纤维化具有良好的耐受性。我们假设高渗透压通过促进新的复合物改变由炎症刺激诱导的选定转录因子的核转位。在本提案期间（2005-2010），我们将首先检查控制原型肺泡细胞因子和 HOsm 修饰的粘附蛋白的 TF 的运输，并研究这些 TF 的复合物，包括与 importins 形成的复合物。 1. 研究 HOsm 改变的 NF-KB 和 IKK 家族成员的易位、它们的 HOsm 改变的细胞质复合物，并通过 IP-MS 蛋白质组学检查 HOsm 促进的替代复合物。 2. 由于 HOsm 不抑制 IL-1 诱导的 p65 Rel A 易位，但会抑制某些细胞因子，因此我们将检查 IRF、C/EBP 的其他重要启动子的易位。和 R/FLAT 家族，并确定他们的 HOsm 改变的伴侣。接下来，为了将调查扩展到已知细胞因子之外，我们将执行： 3. 基因阵列分析，指导潜在转录因子的生物信息学查询。目标是逐步建立一个在 HOSm 期间未成功易位的 TF 库，并确定其隔离伙伴（从 NF-kB 和 IRF 家族成员开始）。最后，我们将转化这些实验室实验来评估雾化 HOsm 对休克动物和创伤患者的耐受性或益处。 4. 评估受创伤动物或接受吸入 HOsm 治疗的患者的肺部炎症和 TF 易位。目标 1-3 的信息将用于解释治疗后的分子表型和 TF 重新分布。