Mechanism of Heat Shock Protein Induction by Glutamine

谷氨酰胺诱导热激蛋白的机制

基本信息

批准号：
8246447
负责人：
PAUL E WISCHMEYER
金额：
$ 26.49万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-05-01 至 2015-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8246447
关键词：
Accident and Emergency department Address Animal Model Animals Binding Biological Assay Cell Line Cell Survival Cell Volumes Cell model Cells Chemicals Clinical Colon Confidential Information Critical Illness Data Enhancers Enzymes Epithelial Cells Evaluation Experimental Models Family Fibroblasts Funding Gene Activation Gene Knock-Out Model Gene Silencing Genes Genetic Transcription Glutamine Goals Health Heat Stress Disorders Heat shock proteins Heat-Shock Proteins 70 Heating Human Inflammatory Injury Instruction Intestines Laboratories Language Link Measures Mediating Metabolism Mission Modeling Nuclear Translocation Operative Surgical Procedures Outcome Pathway interactions Patients Plasmids Promoter Regions Proteins Public Health Reporter Research Research Design Research Methodology Role Sepsis Small Interfering RNA Specificity Stress Techniques Tissues Transactivation Transcription Factor 3 Transcriptional Activation cell injury chemical genetics enzyme pathway glycosylation heat-shock factor 1 ileum improved in vivo inhibitor/antagonist injured promoter protein expression research study tool transcription factor

项目摘要

DESCRIPTION: See instructions. State the application's broad, long-term objectives and specific aims, making reference to the health relatedness ofthe project (i.e., relevance to the mission of the agency). Describe concisely the research design and methods for achieving these goals. Describethe rationale and techniques you will use to pursue these goals.In addition, in two or three sentences, describe in plain, lay language the relevance of this research to public health. If the application is funded, thisdescription, as is, will become public information. Therefore, do not include proprietary/confidential information. Enhanced heat shock protein (HSP) expression protects cells and tissues from injury. Further, enhanced HSP expression improves survival in experimental models of critical illness. However, these findings have not been applied in a clinical setting, as laboratory inducers of HSPs are not safe for human administration. Thus, this powerful tool, that may significantly improve clinical outcome, has yet to be utilized. Our laboratory has shown glutamine (GLN) can safely enhance HSP expression in tissues of critically ill and injured animals and established HSP induction is necessary for GLN's beneficial effect following experimental illness. In a trial of critically ill patients we demonstrated GLN enhanced HSP-70 levels, which correlated with improved outcome. However, the mechanism by which GLN induces HSP expression is unknown. We hypothesize GLN induces HSP expression via activation of the O-linked glycosylation pathway (O-GlcNAc), which is known to depend on GLN as a rate limiting substrate. This pathway can activate key transcription factors required for HSP induction. Our preliminary data indicates GLN increases the activity of the O-GlcNAc pathway and nuclear translocation and activation of key inducers of the HSP pathway, such as Sp1 and heat shock factor-1 (HSF-1). Further, siRNA inhibition of one of the key O-GlcNAc pathway enzymes significantly blunts the GLN-mediated increase in HSP expression. The major focus of this proposal is to determine the mechanism by which GLN induces HSP expression. Our hypothesis is GLN acts via transport into the cell and metabolism by the O-GlcNAc pathway to increase O-linked glycosylation of key transcription factors required for HSP gene activation, which then increases the expression of HSPs. To address this hypothesis, we propose three specific aims: This project will utilize cellular and animal models of illness/injury and employ chemical/genetic inhibition of key enzymes in these pathways to address the following specific aims: 1) Evaluate the role of GLN transport and metabolism via the O-GlcNAc pathway in stress/injury. 2) Evaluate via promoter truncation which promoter regions are key for GLN-mediated HSP expression. Then, determine the effect of GLN on nuclear translocation and transactivation of key transcription factors responsible for HSP expression. 3) Evaluate effects of GLN transport, metabolism, and transcriptional activation on the following cellular and in vivo endpoints: a) HSP expression (multiple families of HSPs), b) tissue/cellular injury, and c) cell volume. This project will elucidate how GLN induces HSP expression in clinical illness and injury. We believe GLN will be able to be administered as a pharmacologic agent prior to surgery or at onset of critical illness/tissue injury (with admittance to ICU/emergency room) to enhance HSP expression and improve survival. This project will elucidate how glutamine induces protective heat shock protein expression in clinical illness and injury. We believe glutamine will be able to be administered as a pharmacologic agent prior to surgery or at onset of critical illness/tissue injury (with admittance to ICU/emergency room) to enhance heat shock protein expression and improve survival.

描述：参见说明。陈述应用程序的广泛、长期目标和具体目标，并提供参考与项目的健康相关性（即与机构使命的相关性）。简明地描述研究实现这些目标的设计和方法。描述您将用于实现这些目标的基本原理和技术。此外，用两到三个句子，用通俗易懂的语言描述这项研究与公共卫生的相关性。如果申请获得资助后，此描述将按原样成为公共信息。因此，不包括专有/机密信息。增强的热休克蛋白 (HSP) 表达可保护细胞和组织免受损伤。进一步，增强了 HSP 表达可提高危重疾病实验模型的存活率。然而，这些发现尚未应用于临床环境，因为实验室的 HSP 诱导剂对人体给药不安全。因此，这种可以显着改善临床结果的强大工具尚未得到利用。我们的实验室已表明谷氨酰胺 (GLN) 可以安全地增强危重和受伤动物组织中的 HSP 表达已确定的 HSP 诱导对于 GLN 在实验性疾病后发挥有益作用是必要的。在一个对危重患者的试验表明，GLN 增强了 HSP-70 水平，这与改善结果。然而，GLN 诱导 HSP 表达的机制尚不清楚。我们假设 GLN 通过激活 O-连接糖基化途径 (O-GlcNAc) 诱导 HSP 表达，该途径是已知依赖 GLN 作为限速底物。该途径可以激活关键转录因子 HSP 诱导所需。我们的初步数据表明 GLN 增加了 O-GlcNAc 的活性 HSP 途径的关键诱导物（例如 Sp1 和热）的核转位和激活休克因子-1 (HSF-1)。此外，siRNA 对关键 O-GlcNAc 途径酶之一的抑制显着抑制 GLN 介导的 HSP 表达增加。该提案的主要重点是确定 GLN 诱导 HSP 表达的机制。我们的假设是 GLN 通过转运进入细胞发挥作用通过 O-GlcNAc 途径进行代谢，以增加关键转录因子的 O 连接糖基化 HSP 基因激活所需的，然后增加 HSP 的表达。为了解决这个假设，我们提出了三个具体目标：该项目将利用疾病/损伤的细胞和动物模型，以及利用这些途径中关键酶的化学/遗传抑制来实现以下具体目标： 1) 评估 GLN 通过 O-GlcNAc 途径转运和代谢在应激/损伤中的作用。 2）评估通过启动子截短，哪些启动子区域是 GLN 介导的 HSP 表达的关键。然后，确定 GLN 对核易位和关键转录因子反式激活的影响热休克蛋白表达。 3) 评估 GLN 转运、代谢和转录激活对以下细胞和体内终点：a) HSP 表达（HSP 的多个家族），b) 组织/细胞损伤，c) 细胞体积。该项目将阐明 GLN 如何在临床疾病和治疗中诱导 HSP 表达受伤。我们相信 GLN 将能够在手术前或发病时作为药物给药严重疾病/组织损伤（进入 ICU/急诊室）以增强 HSP 表达和提高生存率。该项目将阐明谷氨酰胺如何在临床疾病和疾病中诱导保护性热休克蛋白表达受伤。我们相信谷氨酰胺将能够作为药物在手术前或手术时施用危重疾病/组织损伤发作（进入ICU/急诊室）以增强热休克蛋白表达并提高生存率。

项目成果

期刊论文数量（35）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Early propranolol treatment induces lung heme-oxygenase-1, attenuates metabolic dysfunction, and improves survival following experimental sepsis.

DOI：
10.1186/cc12889
发表时间：
2013-09-10
期刊：
Critical care (London, England)
影响因子：
0
作者：
Wilson J;Higgins D;Hutting H;Serkova N;Baird C;Khailova L;Queensland K;Vu Tran Z;Weitzel L;Wischmeyer PE
通讯作者：
Wischmeyer PE

Left ventricular assist device effects on metabolic substrates in the failing heart.

DOI：
10.1371/journal.pone.0060292
发表时间：
2013
期刊：
PloS one
影响因子：
3.7
作者：
Weitzel LB;Ambardekar AV;Brieke A;Cleveland JC;Serkova NJ;Wischmeyer PE;Lowes BD
通讯作者：
Lowes BD

Comment on: probiotic prophylaxis in predicted severe acute pancreatitis: a randomized, double-blind, placebo-controlled trial.

评论：益生菌预防可预测的严重急性胰腺炎：一项随机、双盲、安慰剂对照试验。