hHv1 channels in neutrophils and the innate immune inflammatory response

中性粒细胞中的 hHv1 通道和先天免疫炎症反应

• 批准号: 10677676
• 负责人: Steve A N Goldstein
• 金额: $ 60.06万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-05 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10677676
• 关键词: Acrosome Reaction; Acute; Acute Lung Injury; Acute Respiratory Distress Syndrome; Address; Affinity; Air; Albumins; Anti-Inflammatory Agents; Automobile Driving; Bacteria; Bacterial Pneumonia; Binding; Biophysics; Bronchoalveolar Lavage Fluid; COVID-19; COVID-19 pandemic; Cells; Cessation of life; Chronic; Complex; Data; Development; Disease; Effectiveness; Elastases; Fertilization; Fluorescence Resonance Energy Transfer; Functional disorder; Generations; Goals; Grant; Health; Hospitalization; Host Defense; Human; Hypoxemia; Immune; Immunologic Stimulation; In Vitro; Infection; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Innate Immune System; Ischemic Stroke; Knock-out; Knockout Mice; Leukocytes; Life; Ligands; Liquid substance; Lung; Lung Compliance; Mediating; Medical; Membrane; Methods; Microscopy; Modeling; Morbidity - disease rate; Mus; Oocytes; Oxidants; Oxidases; Pathology; Pathway interactions; Patients; Peptide Hydrolases; Peptides; Phagocytes; Pharmaceutical Preparations; Pharmacology; Pharmacology Study; Pharmacotherapy; Physiology; Pneumonia; Production; Proteins; Protons; Public Health; Publishing; Reactive Oxygen Species; Regulation; Reporting; Research; Respiratory Burst; Role; SARS-CoV-2 infection; Sepsis; Site; Speed; Sperm Capacitation; Structure; Variant; Virus; Wild Type Mouse; Work; base; chronic inflammatory disease; combat; cytokine; design; experience; extracellular; fungus; improved; in vivo; inhibitor; innovation; lung injury; mortality; mouse model; neutrophil; novel; novel strategies; operation; pharmacologic; public health relevance; rational design; response; single molecule; sperm cell; structural biology; success; tool; voltage

SUMMARY/ABSTRACT (30 lines) Relevance to public health. Polymorphonuclear leukocytes (PMN, neutrophils) release reactive oxygen species (ROS) to combat infection, but this inflammatory response can also initiate and propa- gate lung damage. Acute lung injury (ALI) and its severe form, acute respiratory distress syndrome (ARDS) that is fatal in 40% of patients, are characterized by accumulation of albumin-rich fluid in the pulmonary air spaces. Drug therapies focused on downstream cytokine actions have failed to improve morbidity or mortality; we hypothesize, and offer evidence, that targeting the human voltage-gated pro- ton channel (hHv1) at early steps can be more effective. We propose to target hHv1 because (i) the chan- nel in PMN initiates and sustains the inflammatory response, (ii) C6, a unique blocker of hHv1 sup- presses human PMN ROS production, and (iii) C6 suppresses lung compromise in an ALI mouse model. Brief background. This application builds on advances in the last period when we created the first high-affinity and specific direct blocker of hHv1 (C6 peptide) and used it to show, first, that human sperm require hHv1-mediated H+ efflux to initiate capacitation, allowing the acrosomal reaction, and oocyte fertilization and, second, that hHv1 in human PMN is required to produce and sustain release of inflammatory agents, including ROS and proteases, during the innate immune inflammatory response. Unique features and innovation. Our pilot data reveal a second target in the pathway: albumin (Alb) is required to activate hHv1 in human PMN and we describe a peptide (L*) that blocks Alb-activa- tion and ROS production. Supporting our driving hypothesis, we show here that both C6 and L* inhibit hHv1 in human PMN, decreasing ROS production, and that C6 protects in an ALI mouse model, restor- ing lung compliance, and decreasing ROS, proinflammatory cytokines, protein, and PMN in bron- choalveolar lavage fluid. We employ our novel membrane tethered (T-peptide) method to speed struc- ture-function studies and peptide design, show a bivalent C6 (C62) that fully inhibits open hHv1 chan- nels, benefit from advanced biophysical and in vivo methods, and two expert collaborators. Three specific aims. (1) Alb activation of hHv1 seeks the structural and mechanistic basis for the action of Alb and a more potent natural metabolite. (2) Alb regulation of the PMN inflammatory re- sponse seeks to delineate the role of hHv1 in PMN using C6, C62 and L* and the basis for peptide action. (3) Inhibiting acute lung injury with Hv1 inhibitors studies an ALI model in WT and Hv1 KO mice. Significance. This work addresses an unmet medical need, recently made more apparent by the ad- vent of COVID-19-related ALI/ARDS and has broader influence because Hv1 in PMN and other phago- cytes is complicit in additional acute and chronic inflammatory disorders. We propose to apply unique hHv1 inhibitors and innovative methods to understand and suppress this pathophysiology.

摘要/摘要(30行) 与公共卫生的相关性。中性粒细胞(PMN、中性粒细胞)释放反应性 氧物种(ROS)来对抗感染，但这种炎症反应也可以启动和促进- 门肺损伤。急性肺损伤及其严重形式--急性呼吸窘迫综合征 (ARDS)在40%的患者中是致命的，其特征是富含白蛋白的液体在 肺内空气空间。专注于下游细胞因子作用的药物治疗未能改善 发病率或死亡率；我们假设并提供证据，靶向人类电压门控蛋白- 在早期步骤中使用TON通道(HHv1)可能更有效。我们建议以hHv1为目标，因为(I)陈- 中性粒细胞中的NEL启动和维持炎症反应，(Ii)C6，一种独特的hHv1受体阻断剂。 抑制人PMN ROS的产生，以及(Iii)C6抑制ALI小鼠模型的肺损害。 背景简介。此应用程序建立在上一阶段的进展基础上，当时我们创建了第一个 高亲和力和特异性hHv1(C6肽)直接阻滞剂，并用它来显示，首先，人类 精子需要hHv1介导的H+外流来启动获能，从而允许顶体反应，以及 第二，人类中性粒细胞中的hHv1需要产生和维持释放 炎症因子，包括ROS和蛋白酶，在先天免疫炎症反应过程中。 独具特色，勇于创新。我们的试验数据揭示了该途径中的第二个靶点：白蛋白 (白蛋白)是激活人中性粒细胞hHv1所必需的，我们描述了一种阻断白蛋白激活蛋白的多肽(L*)。 TION和ROS生产。支持我们的驾驶假说，我们在这里表明，C6和L*都抑制 在人PMN中的hHv1，减少ROS的产生，以及C6在ALI小鼠模型中的保护作用，恢复- 改善肺顺应性，降低ROS、促炎细胞因子、蛋白质和中性粒细胞。 支气管肺泡灌洗液。我们使用我们的新的膜系留(T-肽)方法来加速结构- 真功能研究和多肽设计表明，二价C6(C62)完全抑制开放的hHv1 Chan- NEL，受益于先进的生物物理和活体方法，以及两位专家合作者。 三个具体目标。(1)hHv1的Alb激活为HHv1的 白蛋白和一种更有效的天然代谢物的作用。(2)白蛋白对中性粒细胞炎症反应的调节 Sensse试图使用C6、C62和L来描述hHv1在PMN中的作用以及多肽作用的基础。 (3)Hv1抑制剂抑制WT和Hv1 KO小鼠急性肺损伤模型的研究。 意义重大。这项工作解决了一个未得到满足的医疗需求，最近的广告更明显地表明了这一点- 新冠肺炎相关的ALI/ARDS的爆发，具有更广泛的影响力，因为PMN和其他吞噬细胞中的HIV-1 细胞是其他急性和慢性炎症性疾病的同谋。我们建议将Unique HHv1抑制剂和了解和抑制这一病理生理学的创新方法。