Physiological vs. pharmacological effects of glucocorticoids on human monocytes.

糖皮质激素对人单核细胞的生理与药理作用。

• 批准号: 8227989
• 负责人: PAUL GUYRE
• 金额: $ 15.8万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-15 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8227989
• 关键词: Acute; Address; Adrenal hormone preparation; Anti-Inflammatory Agents; Anti-inflammatory; Autoimmunity; Behavioral Research; Biological Assay; Biomedical Research; Cells; Clinical; Clinical Protocols; Clinical Research; Data; Development; Dose; Effectiveness; Enzyme-Linked Immunosorbent Assay; Escherichia coli; Evaluation; Event; Exposure to; Flow Cytometry; Fostering; Future; Genes; Glucocorticoids; Health; Human; Hydrocortisone; Immune; Immune response; Immunity; In Vitro; Individual; Individual Differences; Infection; Inflammation; Inflammatory; Inflammatory Response; Interleukin-6; Lead; Leukocytes; Lipopolysaccharides; Measurement; Mediator of activation protein; Messenger RNA; Mitogen-Activated Protein Kinases; Molecular; Mononuclear; Pathway interactions; Phosphorylation; Physiological; Pilot Projects; Play; Production; Regulation; Reporting; Role; Saline; Screening procedure; Steroids; Stress; Testing; Time; Trauma; chemokine receptor; clinically significant; cytokine; design; exposed human population; human study; immune activation; improved; in vivo; innovation; macrophage; monocyte; novel; novel therapeutic intervention; peripheral blood; response

DESCRIPTION (provided by applicant): Pharmacological glucocorticoids (GCs) have been effectively used to treat inflammation for over a half century, resulting in the established paradigm of GC action that addresses only mechanisms that lead to suppression of inflammation and immunity. In contrast, and less appreciated, is the fact that physiological stress-associated concentrations of GCs-but not higher pharmacological levels-play an essential enhancing role in the activation of immune and inflammatory responses. We therefore propose a pilot human study to obtain data that will support a new paradigm, one addressing the mechanisms by which a transient in vivo increase in cortisol to stress-associated levels (e.g., levels that follow trauma) prepares for an enhanced innate immune response when a subsequent activation event occurs (e.g., an infection). Our study is designed to test the hypothesis that preparative phenotypic and molecular changes induced by physiological concentrations of cortisol in human monocytes and macrophages, lead to functionally opposing consequences compared with pharmacological GC doses. Our secondary aim, identifying mechanisms for individual-specific differential responses to 'stress- cortisol' versus 'pharm-cortisol' would, if successful, provide a screening approach with far reaching benefits for nearly all avenues of biomedical, behavioral, and clinical research. Specifically, we propose to: 1. Identify specific cellular and molecular changes that distinguish stress-cortisol enhancement from pharm- cortisol suppression of human monocyte/macrophage innate immune activation pathways. We have found that stress-cortisol pretreatment reproducibly enhances LPS-induced IL-6 production by human monocyte-derived macrophages from some subjects (responders) but not others (non-responders). Inter-individual differences and differential effects of stress- vs. pharm-cortisol will be used to discriminate by flow cytometry, gene profiling analysis and ELISA assays the most relevant molecular events that lead to stress-cortisol enhanced cytokine production. We will specifically interrogate mechanisms by which stress-cortisol enhancement of LPS- induced MAP kinase phosphorylation augments innate immune activation in human macrophages. 2. Identify, at the molecular level, specific monocyte responses that are differentially regulated by stress- cortisol and pharm-cortisol in vivo. Each subject will serve as his/her own control, and will receive 3 in vivo treatments; saline, stress-cortisol (40mg/70kg/6hours), or pharm-cortisol (400mg/70kg/6hours) with an inter- treatment interval that returns measurements to baseline. Flow cytometry, Taqman real time PCR and ELISA assays will be used to determine the in vivo onset, magnitude and durability of stress-cortisol versus pharm- cortisol-induced changes in key cellular regulators of innate immune inflammation. We plan to identify specific molecular events that predict differential in vivo effects of stress-cortisol versus pharm-cortisol, or differences in innate immune activation among individual subjects. Our results will direct future studies to determine important clinical distinctions between anti-inflammatory and pro-inflammatory regulatory effects of GCs. PUBLIC HEALTH RELEVANCE: Steroids related to the natural adrenal hormone hydrocortisone have been used to effectively treat inflammation and autoimmunity for over a half century, but unexplained differences in their effectiveness in different people presents a vexing clinical challenge. Improved understanding of the mechanisms by which these steroids regulate mediators of immunity and inflammation is needed to foster new therapeutic interventions for the treatment of acute systemic inflammation. The studies proposed are novel in that, to the best of our knowledge, we will be the first to examine mechanisms by which physiological stress-associated levels of hydrocortisone enhance immune and inflammatory pathways within human monocytic white blood cells in vivo.

描述（由申请人提供）：半个多世纪以来，药理学糖皮质激素（GC）一直被有效地用于治疗炎症，从而形成了仅针对导致炎症和免疫抑制的机制的既定范例。相比之下，人们较少认识到的事实是，与生理应激相关的 GC 浓度（而不是更高的药理水平）在免疫和炎症反应的激活中发挥着重要的增强作用。因此，我们提出了一项试点人体研究，以获得支持新范式的数据，该范式解决了体内皮质醇短暂增加到压力相关水平（例如创伤后的水平）为增强先天免疫反应做好准备的机制。当后续激活事件发生时（例如感染）。我们的研究旨在检验以下假设：与药理学 GC 剂量相比，人类单核细胞和巨噬细胞中皮质醇生理浓度诱导的制备型表型和分子变化会导致功能上相反的后果。我们的次要目标是确定个体对“压力皮质醇”与“药物皮质醇”的差异反应机制，如果成功的话，将为几乎所有生物医学、行为和临床研究途径提供一种具有深远益处的筛选方法。具体来说，我们建议： 1. 识别特定的细胞和分子变化，以区分压力皮质醇增强和药物皮质醇抑制人类单核细胞/巨噬细胞先天免疫激活途径。我们发现，应激皮质醇预处理可重复增强某些受试者（应答者）而非其他受试者（非应答者）的人单核细胞来源的巨噬细胞 LPS 诱导的 IL-6 产生。应激皮质醇与药物皮质醇的个体间差异和差异效应将用于通过流式细胞术、基因谱分析和 ELISA 测定来区分导致应激皮质醇增强细胞因子产生的最相关分子事件。我们将专门探讨压力皮质醇增强 LPS 诱导的 MAP 激酶磷酸化增强人类巨噬细胞先天免疫激活的机制。 2. 在分子水平上鉴定体内应激皮质醇和药物皮质醇差异调节的特定单核细胞反应。每个受试者将作为自己的对照，并接受3次体内治疗；生理盐水、应激皮质醇（40 毫克/70 公斤/6 小时）或药物皮质醇（400 毫克/70 公斤/6 小时），治疗间间隔可将测量值恢复至基线。流式细胞术、Taqman 实时 PCR 和 ELISA 测定将用于确定体内应激皮质醇与药物皮质醇诱导的先天免疫炎症关键细胞调节因子变化的发生、程度和持久性。我们计划识别特定的分子事件，以预测应激皮质醇与药物皮质醇的体内差异效应，或个体受试者之间先天免疫激活的差异。我们的结果将指导未来的研究，以确定 GC 的抗炎和促炎调节作用之间的重要临床区别。 公共健康相关性：半个多世纪以来，与天然肾上腺激素氢化可的松相关的类固醇已被用于有效治疗炎症和自身免疫，但其在不同人群中的疗效存在无法解释的差异，这提出了令人烦恼的临床挑战。需要更好地了解这些类固醇调节免疫和炎症介质的机制，以培育治疗急性全身炎症的新治疗干预措施。所提出的研究是新颖的，据我们所知，我们将是第一个研究生理应激相关的氢化可的松水平增强人单核白细胞体内免疫和炎症途径的机制。