THE MOLECULAR REGULATION OF NEUTROPHIL CALCIUM SIGNALING IN ACUTE LUNG INJURY

急性肺损伤中性粒细胞钙信号传导的分子调控

• 批准号: 9465405
• 负责人: Regina Clemens
• 金额: $ 17.31万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9465405
• 关键词: Accounting; Acute Lung Injury; Address; Alveolar; Animals; Bacteria; Biological Models; CRISPR/Cas technology; Calcium; Calcium Channel; Calcium Signaling; Cells; Cessation of life; Clinical; Cytoplasmic Granules; Data; Disease; Endoplasmic Reticulum; Enzymes; Epithelial; Etiology; Functional disorder; Future; Genes; Goals; Hematopoietic stem cells; Host Defense; Human; Hypoxemia; Immune; Immunodeficient Mouse; In Vitro; Inflammatory; Inflammatory Response; Injury; Irrigation; Knockout Mice; Knowledge; Lead; Lymphocyte; Mediating; Mediator of activation protein; Medical; Mission; Modeling; Molecular; Morbidity - disease rate; Morphology; Multiple Trauma; Mus; Mutant Strains Mice; National Institute of Allergy and Infectious Disease; Neutrophil Activation; Pathogenesis; Pathway interactions; Patients; Persons; Positioning Attribute; Process; Production; Proteins; Publishing; Pulmonary Edema; Pulmonary Pathology; Reactive Oxygen Species; Reagent; Regulation; Reperfusion Therapy; Research; Residual state; Rest; Role; STIM1 gene; Severity of illness; Signal Pathway; Signal Transduction; Sterility; Supportive care; System; Techniques; Testing; Tissues; Toxic Neutrophil; Translating; United States National Institutes of Health; base; cytokine; extracellular; humanized mouse; in vivo; insight; liver ischemia; lung development; lung injury; mortality; neutrophil; novel; novel strategies; novel therapeutic intervention; public health relevance; release of sequestered calcium ion into cytoplasm; response; sensor; therapeutic target

 DESCRIPTION (provided by applicant): Acute lung injury (ALI) is a devastating disease characterized by alveolar-epithelial barrier disruption and resultant pulmonary edema and hypoxemia. There are currently no effective disease-modifying therapies and supportive care remains the mainstay of medical management. Ultimately, ALI leads to multi-organ dysfunction and/or death in up to 30% of patients. Neutrophils are clearly important for host defense against bacteria, however toxic neutrophil mediators such as reactive oxygen radicals, granule enzymes and neutrophil extracellular traps also contribute to the pathogenesis of lung injury by causing endothelial, parenchymal and alveolar injury. Neutrophils are prime targets for manipulating the inflammatory response and therefore it is critical to understand the molecular mechanisms that guide neutrophil responses. Dr. Clemens' long-term research goal is to understand how intracellular signaling pathways regulate neutrophil activation during acute lung injury and other inflammatory diseases. Calcium is an evolutionarily conserved signaling messenger that is a central component of multiple signaling pathways. In immune cells, increases in cytoplasmic calcium are controlled via store operated calcium entry (SOCE), where calcium release-activated calcium (CRAC) channels allow influx of extracellular calcium when endoplasmic reticulum (ER) stores are depleted. STIM proteins are ER calcium "sensors" which sense calcium depletion and directly gate ORAI, a pore subunit of the CRAC channel. Little is known about STIM, ORAI or the molecular regulation of calcium signaling in neutrophils. The objectives of this proposal are to define the molecular pathways that regulate SOCE in neutrophils and to determine how these pathways modulate neutrophil activation during ALI. The central hypotheses are: 1) STIM calcium sensors and ORAI calcium channels cooperate to regulate neutrophil calcium signaling, and 2) calcium-dependent neutrophil activation is critical for induction of ALI. The studies proposed in Aims 1 and 2 will identify the molecular machinery required for SOCE in neutrophils and determine the role of neutrophil calcium signaling in acute lung injury using a unique set of mice with neutrophil specific deletion in Stim or Orai genes. Aim 3 will utilize a novel approach with humanized mice to test these hypotheses in human neutrophils. This project is relevant to the missions of the NIH and NIAID since these studies will elucidate previously unexplored pathways of calcium signaling in neutrophils that may lead to new therapeutic approaches to treat acute lung injury and other inflammatory processes.

 描述（由申请人提供）：急性肺损伤（ALI）是一种破坏性疾病，其特征是肺泡上皮屏障破坏以及由此导致的肺水肿和低氧血症。目前尚无有效的疾病缓解疗法，支持性护理仍然是医疗管理的支柱。最终，ALI 会导致高达 30% 的患者多器官功能障碍和/或死亡。中性粒细胞对于宿主抵御细菌的防御显然很重要，然而，有毒的中性粒细胞介质，如活性氧自由基、颗粒酶和中性粒细胞胞外陷阱，也通过引起内皮、实质和肺泡损伤，从而促进肺损伤的发病机制。中性粒细胞是操纵炎症反应的主要目标，因此了解指导中性粒细胞反应的分子机制至关重要。 Clemens 博士的长期研究目标是了解细胞内信号通路如何在急性肺损伤和其他炎症性疾病期间调节中性粒细胞激活。钙是一种进化上保守的信号传导信使，是多种信号传导途径的核心组成部分。在免疫细胞中，细胞质钙的增加是通过钙库操作的钙进入（SOCE）来控制的，其中钙释放激活的钙（CRAC）通道允许当内质网（ER）储备耗尽时细胞外钙的流入。 STIM 蛋白是 ER 钙“传感器”，可感知钙消耗并直接门控 ORAI（CRAC 通道的孔亚基）。人们对 STIM、ORAI 或中性粒细胞中钙信号传导的分子调节知之甚少。该提案的目的是定义调节中性粒细胞 SOCE 的分子途径，并确定这些途径如何在 ALI 期间调节中性粒细胞激活。中心假设是：1) STIM 钙传感器和 ORAI 钙通道协同调节中性粒细胞钙信号传导，2) 钙依赖性中性粒细胞激活对于诱导 ALI 至关重要。目标 1 和 2 中提出的研究将确定中性粒细胞中 SOCE 所需的分子机制，并使用一组具有 Stim 或 Orai 基因中性粒细胞特异性缺失的独特小鼠来确定中性粒细胞钙信号传导在急性肺损伤中的作用。目的 3 将利用一种新的方法，用人源化小鼠来测试人类中性粒细胞的这些假设。该项目与 NIH 和 NIAID 的使命相关，因为这些研究将 阐明以前未探索的中性粒细胞中钙信号传导途径，这可能会导致治疗急性肺损伤和其他炎症过程的新治疗方法。