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是针对入侵病原体的宿主防御的主要参与者。相反，过量的IL-1β产生 和/或激活可以确定为系统，从而导致不需要的组织注射和夸张的组织注射。 因此，需要很好地控制IL-1β的生物活性。传统上，IL-1β调节的机制具有 专注于模式识别受体诱导的基因转录和炎症体介导的裂解 Pro-IL-1β。完整的IL-1β细胞因子调节曲目仍然很大程度上未知。目的 拟议的研究是识别和表征调节成熟IL-生物活性的生化事件 1β从细胞释放后。最近，我们发现高度保守的半胱氨酸S-谷胱甘肽化 IL-1β中的Cys-188居住地通过防止其不可逆的活性氧来正调节IL-1β的生物活性 物种（ROS）吸收的氧化和失活。蛋白质谷胱甘肽化是动态和可逆的。我们 进一步证明，谷胱甘肽1（GRX1）是一种催化丙二酰二酰化的酶，并且存在 活跃在血清和BM的细胞外空间中，并物理调节IL-1β谷胱甘肽。 总之，这些结果导致我们假设ROS诱导的半胱氨酸S-谷胱甘肽及其 GRX1的调节是在病理生理下控制IL-1β活性的关键调节机制 状况。在当前的研究中，我们将在IL-1β活性为临床相关模型中检验这一假设 辐照后，必不可少的和足够的造血系统有效恢复。首先，我们会的 确定半胱氨酸s-谷胱甘肽化在确定内生产生的IL-1β的生物活性中的作用 在辐射后骨髓（BM）恢复期间（AIM I）。此外，我们将阐明Grx1在 调节辐照小鼠的BM恢复中内源产生的IL-1β的S-谷二酰化。我们也会 识别在BM中产生GRX1的细胞类型（AIM II）。最后，我们将尝试加速BM恢复 通过靶向IL-1βS-谷胱甘肽的辐照。我们将首次检查谷胱甘肽是否静脉注射 （GSH IV）治疗可以提高IL-1β生物活性并加速造血系统的恢复 辐照小鼠。此外，我们将研究IL-1β188C/s是否是一种不可能是IL-1β的突变形式 与WT IL-相比 1β（AIM III）。总之，这三个特定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-谷胱甘肽在控制体内IL-1β活性中的作用在临床相关 设置。走向转化研究范式，这项研究的结果将有助于我们确定新颖 治疗靶标（例如ROS，GRX1和相关途径），用于加速接受BM的患者的BM恢复 放疗。