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

基本信息

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
项目类别：
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

项目摘要

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。多形核白细胞（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传播门肺损伤。急性肺损伤 (ALI) 及其严重形式，急性呼吸窘迫综合征 (ARDS) 导致 40% 的患者致命，其特征是富含白蛋白的液体在体内积聚肺空气空间。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；我们假设并提供证据，针对人类电压门控亲早期的 ton 通道 (hHv1) 可能会更有效。我们建议以 hHv1 为目标，因为 (i) PMN 中的 nel 启动并维持炎症反应，(ii) C6，hHv1 支持的独特阻断剂抑制人类 PMN ROS 的产生，(iii) C6 抑制 ALI 小鼠模型中的肺损伤。简要背景。该应用程序建立在我们创建第一个应用程序的上一时期的进步基础上 hHv1（C6 肽）的高亲和力和特异性直接阻断剂，并用它首先表明，人类精子需要 hHv1 介导的 H+ 流出来启动获能，从而发生顶体反应，并且其次，人类 PMN 中的 hHv1 是产生和持续释放先天免疫炎症反应过程中的炎症因子，包括 ROS 和蛋白酶。独特的功能和创新。我们的试验数据揭示了该途径中的第二个目标：白蛋白 (Alb) 是激活人类 PMN 中 hHv1 所必需的，我们描述了一种肽 (L*)，它可以阻断 Alb-activa- 化和ROS的产生。为了支持我们的驱动假设，我们在此表明 C6 和 L* 都抑制 hHv1 存在于人类 PMN 中，减少了 ROS 的产生，C6 在 ALI 小鼠模型中起到保护作用，恢复了增强肺顺应性，并减少支气管中的 ROS、促炎细胞因子、蛋白质和中性粒细胞 (PMN) 胆泡灌洗液。我们采用新型膜栓系（T 肽）方法来加速结构真实功能研究和肽设计表明，二价 C6 (C62) 可以完全抑制开放 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激活寻找其结构和机制基础 Alb 和更有效的天然代谢物的作用。 (2) Alb对PMN炎症反应的调节 sponse 试图使用 C6、C62 和 L* 以及肽作用的基础来描述 hHv1 在 PMN 中的作用。 (3) 用 Hv1 抑制剂抑制急性肺损伤研究 WT 和 Hv1 KO 小鼠的 ALI 模型。意义。这项工作解决了未满足的医疗需求，最近通过广告变得更加明显是 COVID-19 相关 ALI/ARDS 的发源地，并且具有更广泛的影响，因为 PMN 和其他噬菌体中的 Hv1 细胞是其他急性和慢性炎症性疾病的同谋。我们建议应用独特的 hHv1 抑制剂和创新方法来了解和抑制这种病理生理学。