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Hypothermia enhances inflammatory cytokine expression via NF-kappa B

低温通过 NF-κ B 增强炎症细胞因子的表达

基本信息

批准号：
7921021
负责人：
KAREN D FAIRCHILD
金额：
$ 13.01万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-11 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7921021
关键词：
Acetylation Adolescent Adverse effects Biological Assay Brain Injuries Child Childhood Chromatin Clinical Clinical Trials Critical Care Cytokine Activation Cytokine Gene Data Deacetylase Event Gene Expression Genetic Transcription Goals Histone H3 Human IL8 gene Immunoblotting In Vitro Inflammation Inflammatory Investigation Mediator of activation protein Molecular Mus NF-kappa B Neonatal Outcome Patients Phosphoric Monoester Hydrolases Phosphorylation Production Protein phosphatase Protocols documentation RNA Interference Reporter Reporter Genes Research Research Personnel Role System Testing Therapeutic Transactivation Transgenic Organisms Traumatic Brain Injury chromatin immunoprecipitation cytokine cytotoxic histone deacetylase 3 in vitro activity in vivo insight lentiviral-mediated macrophage natural hypothermia neonatal hypoxic-ischemic brain injury neonate overexpression p65 prevent programs promoter research study transcription factor

项目摘要

DESCRIPTION (provided by applicant): The goal of the proposed research is for Dr. Fairchild to transition into an independent investigator elucidating the cellular, molecular, and in vivo effects of hypothermia on NF kappa B-dependent gene expression. Therapeutic hypothermia (HT) is under investigation in children with traumatic brain injury and neonates with hypoxic ischemic encephalopathy, and there are many other potential uses of this therapy in pediatric and neonatal critical care. While HT appears to be neuroprotective in some patients, concerns remain over potential detrimental effects. Dr. Fairchild has shown that HT in the range used in clinical trials increases activation of the transcription factor nuclear factor kappa B (NF-/kB) and expression of the pro- inflammatory cytokines TNFa, IL-1/3, and IL-8 in LPS-stimulated human macrophages. When overexpressed, these cytokines can be cytotoxic. Preliminary experiments in juvenile mice show that in vivo hypothermia also significantly alters cytokine production. Our overall goal is to understand the mechanisms of hypothermic enhancement of NF-/kB-dependent gene expression in order to better anticipate and perhaps prevent adverse effects of therapeutic HT. Our preliminary data show that HT alters key phosphorylation and acetylation events which favor association of the p65 transactivation component of NF-/kB with cytokine gene promoters on chromatin. In Aim 1 we will elucidate the role of altered NF-/kB p65 transactivation in the hypothermic enhancement of cytokine production, using Gal4-p65 reporter gene studies and lentiviral mediated RNA interference. Experiments in Aim 2 will determine how alterations in critical phosphorylation and acetylation events contribute to the HT effect on pro-inflammatory cytokines, through immunoblotting, in vitro phosphatase and deacetylase assays, and RNA interference. Using chromatin immunoprecipitation, we will elucidate effects of HT on loading of phosphorylated and acetylated p65 and its coactivators and compressors on specific cytokine gene promoters. Finally, in Aim 3, we will develop a system for studying in vivo effects of HT on NF-/kB activation and cytokine expression in juvenile transgenic NF-/kB reporter mice. HT is under investigation as a neuroprotective therapy for children and neonates with brain injury, and many other clinical uses of HT have been proposed. Ultimately, our studies may provide insight into appropriate indications, protocols, and adjunct strategies to optimize outcome of patients undergoing therapeutic HT.

描述（由申请人提供）：提议的研究目标是让Fairchild博士转变为一名独立研究者，阐明低温对NF κ b依赖基因表达的细胞、分子和体内影响。治疗性低温（HT）在创伤性脑损伤儿童和新生儿缺氧缺血性脑病中的应用正在研究中，并且在儿科和新生儿重症监护中有许多其他潜在的应用。虽然HT对某些患者似乎具有神经保护作用，但对潜在有害影响的担忧仍然存在。Fairchild博士已经证明，在临床试验中使用的范围内，HT增加了转录因子核因子κ B （NF-/kB）的激活以及lps刺激的人巨噬细胞中促炎细胞因子TNFa、IL-1/3和IL-8的表达。当过度表达时，这些细胞因子可能具有细胞毒性。幼鼠的初步实验表明，体内低温也显著改变细胞因子的产生。我们的总体目标是了解低温增强NF-/ kb依赖性基因表达的机制，以便更好地预测和预防治疗性高温疗法的不良反应。我们的初步数据表明，高温改变了关键的磷酸化和乙酰化事件，这些事件有利于NF-/kB的p65反活化组分与染色质上的细胞因子基因启动子的关联。在Aim 1中，我们将利用Gal4-p65报告基因研究和慢病毒介导的RNA干扰，阐明NF-/kB p65转激活改变在低温增强细胞因子产生中的作用。Aim 2的实验将通过免疫印迹、体外磷酸酶和去乙酰化酶测定以及RNA干扰，确定关键磷酸化和乙酰化事件的改变如何影响HT对促炎细胞因子的影响。利用染色质免疫沉淀，我们将阐明高温对磷酸化和乙酰化p65及其协同激活因子和压缩因子在特定细胞因子基因启动子上的负载的影响。最后，在Aim 3中，我们将开发一个系统来研究HT对转基因NF-/kB幼年小鼠体内NF-/kB活化和细胞因子表达的影响。目前正在研究HT作为儿童和新生儿脑损伤的神经保护疗法，并且已经提出了许多其他HT的临床应用。最终，我们的研究可能为适当的适应症、方案和辅助策略提供见解，以优化接受治疗性HT的患者的结果。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

FDG-PET Quantification of Lung Inflammation with Image-Derived Blood Input Function in Mice.

DOI：
10.1155/2011/356730
发表时间：
2011
期刊：
International journal of molecular imaging
影响因子：
0
作者：
Locke LW;Williams MB;Fairchild KD;Zhong M;Kundu BK;Berr SS
通讯作者：
Berr SS

Cytokine screening identifies NICU patients with Gram-negative bacteremia.

DOI：
10.1038/pr.2011.45
发表时间：
2012-03
期刊：
PEDIATRIC RESEARCH
影响因子：
3.6
作者：
Raynor, Laura L.;Saucerman, Jeffrey J.;Akinola, Modupeola O.;Lake, Douglas E.;Moorman, J. Randall;Fairchild, Karen D.
通讯作者：
Fairchild, Karen D.

Hypothermia increases interleukin-6 and interleukin-10 in juvenile endotoxemic mice.