Regulation of IL-1β bioactivity by Cysteine S-glutathionylation

半胱氨酸 S-谷胱甘肽化调节 IL-1β 生物活性

基本信息

批准号：
10620756
负责人：
Hongbo R Luo
金额：
$ 16.53万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-12 至 2024-04-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10620756
关键词：
Acceleration Affect Biochemical Bone Marrow Bone Marrow Cells Bone Marrow Purging Cells Cysteine Dose Limiting Enzymes Event Extracellular Space Generations Genetic Transcription Glutathione Glutathione Disulfide Grx1 protein Hematopoietic System Host Defense Hydrogen Peroxide In Vitro Infection Inflammasome Inflammation Interleukin-1 beta Invaded Mass Spectrum Analysis Measures Mediating Modeling Modification Mus Pathway interactions Patients Pattern recognition receptor Physiological Play Post-Translational Protein Processing Production Protein S Proteins Radiation Toxicity Radiation therapy Reactive Oxygen Species Recovery Regulation Reporting Research Role Serum Stress Sulfinic Acids Sulfonic Acids System Testing Therapeutic Intervention Thiol Disulfide Oxidoreductase Tissues Translational Research Up-Regulation cell type clinical application clinically relevant cysteine sulfinic acid cytokine disulfide bond experimental study extracellular gamma irradiation glutaredoxin improved in vivo intravenous injection irradiation mutant new therapeutic target novel novel therapeutic intervention oxidation pathogen prevent protective effect protein B thioltransferase

项目摘要

Project Summary IL-1β is a major player in host defense against invading pathogens. Conversely, excessive IL-1β production and/or activation can be detrimental to the system, resulting in unwanted and exaggerated tissue inflammation. Hence, the bioactivity of IL-1β needs to be well controlled. Mechanisms of IL-1β regulation have traditionally focused on pattern recognition receptor-induced gene transcription and inflammasome-mediated cleavage of pro-IL-1β. The complete IL-1β cytokine regulatory repertoire is still largely unknown. The objective of the proposed research is to identify and characterize biochemical events that modulate the bioactivity of mature IL- 1β after its release from the cells. We recently found that cysteine S-glutathionylation of the highly conserved Cys-188 residue in IL-1β positively regulates IL-1β bioactivity by preventing its irreversible reactive oxygen species (ROS)-elicited oxidation and deactivation. Protein glutathionylation is dynamic and reversible. We further demonstrated that Glutaredoxin 1 (Grx1), an enzyme that catalyzes deglutathionylation, is present and active in the extracellular space in serum and BM, and physiologically regulates IL-1β glutathionylation. Together, these results lead us to hypothesize that ROS-induced cysteine S-glutathionylation and its modulation by Grx1 are key regulatory mechanisms controlling IL-1β activity under pathophysiological conditions. In current study, we will test this hypothesis in a clinically relevant model in which IL-1β activity is both essential and sufficient for efficient recovery of the hematopoietic system after irradiation. First, we will determine the role of cysteine S-glutathionylation in regulating the bioactivity of endogenously produced IL-1β during bone marrow (BM) recovery after irradiation (Aim I). In addition, we will elucidate the function of Grx1 in regulating S-glutathionylation of endogenously produced IL-1β in BM recovery in irradiated mice. We will also identify the cell types that produce Grx1 in the BM (Aim II). Finally, we will try to accelerate BM recovery after irradiation by targeting IL-1β S-glutathionylation. We will first examine whether glutathione intravenous injection (GSH IV) therapy can elevate IL-1β bioactivity and accelerate the recovery of the hematopoietic system in irradiated mice. In addition, we will investigate whether IL-1β 188C/S, a mutant form of IL-1β that can not be oxidized and deactivated, is more potent in eliciting BM protective effect in irradiated mice compared to WT IL- 1β (Aim III). Together, experiments proposed in these three specific aims will provide a better understanding of the role of ROS-induced cysteine S-glutathionylation in controlling IL-1β activity in vivo in clinically relevant settings. Toward the translational research paradigm, results from this study will assist us to identify novel therapeutic targets (e.g. ROS, Grx1, and related pathways) for accelerating BM recovery in patients receiving radiotherapy.

项目概要 IL-1β 是宿主防御离线、过量 IL-1β 产生的主要因素。和/或激活可能会损害系统，导致不必要的和夸大的组织炎症。因此，传统上需要很好地控制IL-1β的生物活性。专注于模式识别受体诱导的基因转录和炎症介导的裂解完整的 IL-1β 细胞因子调节库仍很大程度上未知。拟议的研究旨在识别和表征调节成熟IL-生物活性的生化事件 1β从细胞中释放后，我们最近发现半胱氨酸的S-谷胱甘肽化高度保守。 IL-1β 中的 Cys-188 残基通过阻止其不可逆活性氧来正向调节 IL-1β 生物活性物种（ROS）引起的氧化和失活是动态的和可逆的。进一步证明谷氧还蛋白 1 (Grx1)（一种催化去谷胱甘肽化的酶）的存在并且在血清和 BM 的细胞外空间中活跃，并在生理上调节 IL-1β 谷胱甘肽化。总之，这些结果使我们能够竞争 ROS 诱导的半胱氨酸 S-谷胱甘肽化及其 Grx1 的调节是病理生理条件下控制 IL-1β 活性的关键调节机制在当前的研究中，我们将在临床相关模型中检验这一假设，其中 IL-1β 活性为对于辐射后造血系统的有效恢复既必要又充分。确定半胱氨酸S-谷胱甘肽化在调节内源产生的IL-1β生物活性中的作用此外，我们将阐明 Grx1 在照射后骨髓 (BM) 恢复期间的功能。我们还将在受辐射小鼠的 BM 恢复中调节内源性 IL-1β 的 S-谷胱甘肽化。确定 BM 中产生 Grx1 的细胞类型（目标 II）。通过靶向IL-1β S-谷胱甘肽的照射我们首先检查是否静脉注射谷胱甘肽。 (GSH IV)治疗可提高IL-1β生物活性并加速造血系统的恢复此外，我们将研究IL-1β 188C/S（IL-1β的突变形式）是否不能被照射。被氧化和失活，与 WT IL- 相比，在激发受辐射小鼠的 BM 保护作用方面更有效 1β（目标 III）。这三个具体目标中提出的实验将提供更好的理解。 ROS诱导的半胱氨酸S-谷胱甘肽化在临床相关体内控制IL-1β活性中的作用对于转化研究范式，这项研究的结果将帮助我们识别新颖的内容。加速接受治疗的患者的 BM 恢复的治疗靶点（例如 ROS、Grx1 和相关途径）放射治疗。