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

• 批准号: 10405596
• 负责人: Hongbo R Luo
• 金额: $ 44.25万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-12 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10405596
• 关键词: 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; Lead; Mass Spectrum Analysis; Measures; Mediating; Modeling; Modification; Mus; Oxides; 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

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.

项目摘要 IL-1β是宿主防御入侵病原体的主要分子。相反，过多的IL-1β产生 和/或激活可能对系统有害，导致不想要的和夸大的组织炎症。 因此，需要很好地控制IL-1β的生物活性。传统上，IL-1β的调节机制 重点研究了模式识别受体诱导的基因转录和炎性小体介导的切割 前白介素1β。完整的IL-1β细胞因子调控谱在很大程度上仍不清楚。该计划的目标是 拟议的研究是为了识别和表征调节成熟IL-2生物活性的生化事件。 从细胞中释放后1β。我们最近发现，S的半胱氨酸-谷胱甘肽高度保守 IL-1β中Cys-188残基通过保护不可逆的活性氧正向调节IL-1β的生物活性 物种(ROS)--引发氧化和失活。蛋白质的谷胱甘肽基化是动态的和可逆的。我们 进一步证明存在谷氧化还蛋白1(Grx1)，这是一种催化谷胱甘肽脱硫基化的酶。 活跃于血清和骨髓的胞外间隙，并在生理上调节IL-1β的谷胱甘肽基化。 综上所述，这些结果导致我们假设ROS诱导的半胱氨酸S谷胱甘肽基化和它的 GRX1的调控是病理生理条件下控制IL-1β活性的关键机制 条件。在目前的研究中，我们将在一个临床相关的模型中检验这一假设，在该模型中，IL-1β活性是 既是辐射后有效恢复造血系统所必需的，也是充分的。首先，我们将 半胱氨酸S谷胱甘肽基化在调节内源性IL-1β生物活性中的作用 在照射后骨髓(BM)恢复过程中(目标I)。此外，我们还将阐明Grx1在 辐射后小鼠骨髓恢复过程中内源性IL-1β对S谷胱甘肽的调节作用我们还将 确定BM中产生Grx1的细胞类型(AIM II)。最后，我们将努力加快黑石恢复 照射靶向IL-1βS-谷胱甘肽。我们首先要检查一下静脉注射谷胱甘肽 谷胱甘肽IV治疗可提高IL-1β活性，加速造血系统的恢复 受过辐射的小鼠。此外，我们还将调查IL-1β188C/S，IL-1β的突变形式是否不能 与WT IL-1相比，氧化失活能更有效地诱导照射小鼠骨髓保护作用。 1β(AIM III)。总之，在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-谷胱甘肽基化在体内控制IL-1β活性的作用 设置。对于翻译研究范式，本研究的结果将有助于我们识别小说 促进骨髓恢复的治疗靶点(如ROS、Grx1和相关通路) 放射治疗。