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: 10521974
• 负责人: Steve A N Goldstein
• 金额: $ 64.23万
• 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/10521974
• 关键词: 2019-nCoV; 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; Immunization; 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; Oxides; 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; Sepsis; Site; Speed; Sperm Capacitation; Structure; Variant; Vent; 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; peptide L; 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） 与公共卫生有关。多形核白细胞（PMN，中性粒细胞）释放反应性 氧气（ROS）以对抗感染，但是这种炎症反应也可以引发和促进 门肺损伤。急性肺损伤（ALI）及其严重形式，急性呼吸窘迫综合征 （ARDS）在40％的患者中致命的（ARDS）的特征是富含白蛋白的液体在 肺气空间。专注于下游细胞因子作用的药物疗法未能改善 发病率或死亡率；我们假设并提供了证据，表明针对人类电压门控促进 早期步骤的吨通道（HHV1）可能更有效。我们建议针对HHV1，因为（i）chan- NEL在PMN中启动并维持炎症反应，（ii）C6，这是HHV1 SUP-的独特阻滞剂 按下人类PMN ROS的产生，（iii）C6抑制了ALI小鼠模型中的肺部妥协。 简短背景。该应用程序建立在我们创建第一个的最后一个时期的进步上 HHV1（C6肽）的高亲和力和特定的直接阻滞剂，首先用它来表明人类 精子需要HHV1介导的H+外排才引发电容，允许粘合症反应，并且 卵母细胞受精，其次，人类PMN中的HHV1需要生产和维持释放 先天免疫炎症反应期间，包括ROS和蛋白酶在内的炎症剂。 独特的功能和创新。我们的飞行员数据在途径中揭示了第二个目标：白蛋白 （ALB）需要在人PMN中激活HHV1，我们描述了阻断Alb-Activa-的肽（L*） 产生和ROS的产生。支持我们的驾驶假设，我们在这里表明C6和L*都可以抑制 HHV1在人PMN中，降低ROS的产生，C6在Ali小鼠模型中保护，还原 肺合顺序性，降低ROS，促炎细胞因子，蛋白质和PMN 选择性灌洗液。我们采用新型的膜束缚（T肽）方法来加快结构 特鲁功能研究和肽设计显示了二价C6（C62），该C6（C62）完全抑制开放的HHV1 Chan- NELS受益于先进的生物物理和体内方法，以及两名专家合作者。 三个具体目标。 （1）HHV1的ALB激活寻求结构和机械基础 ALB和更有效的天然代谢产物的作用。 （2）PMN炎症性重新调节 Sponse试图使用C6，C62和L*来描述HHV1在PMN中的作用，以及肽作用的基础。 （3）使用HV1抑制剂抑制急性肺损伤研究WT和HV1 KO小鼠中的ALI模型。 意义。这项工作解决了未满足的医疗需求，最近使广告变得更加明显 COVID-19与ALI/ARDS的通风口，并且具有更大的影响，因为HV1在PMN和其他Phago- 细胞与其他急性和慢性炎症性疾病相同。我们建议应用独特 HHV1抑制剂和创新方法可以理解和抑制这种病理生理学。