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.

项目概要 IL-1β 是宿主防御入侵病原体的主要参与者。相反，IL-1β 产生过多 和/或激活可能对系统有害，导致不必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。 IL-1β 的调节机制传统上被认为是 专注于模式识别受体诱导的基因转录和炎性体介导的裂解 IL-1β原。完整的 IL-1β 细胞因子调节指令在很大程度上仍然未知。该计划的目标 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。我们最近发现高度保守的半胱氨酸S-谷胱甘肽化 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）引起氧化和失活。蛋白质谷胱甘肽化是动态且可逆的。我们 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，这些结果使我们推测 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 状况。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来说是必要且充分的。首先，我们将 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 照射后骨髓 (BM) 恢复期间（目标 I）。此外，我们将阐明 Grx1 的功能 在受辐射小鼠的 BM 恢复中调节内源产生的 IL-1β 的 S-谷胱甘肽化。我们还将 识别 BM 中产生 Grx1 的细胞类型（目标 II）。最后，我们会尽力加速BM恢复 通过靶向 IL-1β S-谷胱甘肽进行照射。我们首先检查是否可以静脉注射谷胱甘肽 (GSH IV)治疗可提高IL-1β生物活性并加速造血系统的恢复 受辐射的老鼠。此外，我们将研究IL-1β 188C/S（IL-1β的突变形式）是否不能被 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）。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-谷胱甘肽化在临床​​相关体内控制IL-1β活性中的作用 设置。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 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。