The role of adenosine in the pathogenesis of influenza

腺苷在流感发病机制中的作用

• 批准号: 8298185
• 负责人: IAN CHRISTOPHER DAVIS
• 金额: $ 37.74万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8298185
• 关键词: 5' -Nucleotidase; ATP Synthesis Pathway; Acute; Acute Lung Injury; Adenosine; Adenosine A1 Receptor; Award; Biochemical; Biological Assay; Bone Marrow; C57BL/6 Mouse; Catabolism; Cell physiology; Cells; Cessation of life; Data; Development; Disease; Epithelial Cells; Extracellular Signal Regulated Kinases; Functional disorder; Funding; Generations; Health; Human; Hypoxemia; Inbred BALB C Mice; Infection; Inflammatory; Influenza; Influenza A virus; Injury; Ion Transport; Knockout Mice; Lead; Life; Liquid substance; Lung; Lung diseases; Measures; Mediating; Mediator of activation protein; Methodology; Modeling; Mus; NF-kappa B; National Center for Research Resources; Nucleotides; Outcome; Pathogenesis; Patients; Play; Publications; Pulmonary Edema; Purinergic P1 Receptors; Receptor Signaling; Research; Respiratory Syncytial Virus Infections; Respiratory physiology; Respiratory syncytial virus; Role; Signal Transduction; Sorting - Cell Movement; Symptoms; Testing; Therapeutic Intervention; Translating; Viral; Virus; Virus Diseases; Work; adenosine receptor activation; alveolar type II cell; base; career; combat; extracellular; improved; infancy; influenza epidemic; influenzavirus; innovation; lung development; lung injury; man; neutrophil; novel; nucleotide metabolism; pandemic disease; public health relevance; purine metabolism; receptor expression; respiratory; seasonal influenza; therapeutic target; tool; virology

DESCRIPTION (provided by applicant): Influenza A viruses cause a highly contagious acute respiratory disease. Seasonal influenza epidemics cause >300,000 deaths/yr worldwide, while pandemics elicited devastating loss of life in the 20th century, and may do so in the 21st. Currently, however, essential mechanisms underlying development of lung dysfunction and injury in influenza or any other pulmonary viral infection are poorly defined. Both respiratory syncytial virus (RSV) and influenza virus infection of BALB/c mice induce increased channel-mediated release of the nucleotides UTP and ATP into the bronchoalveolar lining fluid (BALF), and, in both infections, elevated BALF UTP contributes to development of lung edema and hypoxemia. RSV infection in BALB/c mice causes only mild disease while influenza causes more severe disease with many of the pathophysiologic features of acute lung injury, which is also seen in lethal pandemic or epidemic influenza in man. Importantly, following influenza, but not RSV, infection, elevated ATP release into BALF is accompanied by increased activation of A1-subtype adenosine receptors (AdoR) by the ATP degradation product adenosine. Adenosine stimulation of neutrophil A1-AdoR plays a significant role in the initiation and promotion of acute lung injury. The central hypothesis of this proposal is that influenza stimulates de novo ATP synthesis and release from infected ATII cells, that released ATP is metabolized to adenosine at an accelerated rate (due to increased ectonucleotidase CD73 activity), and that, by its effects on A1-AdoR, adenosine in the BALF plays a pivotal role in inducing lung injury in influenza. This hypothesis will be tested in 3 Specific Aims: 1) To determine temporal effects of influenza infection on pulmonary nucleotide metabolism, changes in nucleotide metabolism following influenza infection of MLE-12 cells and purified alveolar type II (ATII) cells, as well as FACS-sorted ATII cells from C57Bl/6 or SP- C/GFP mice will be measured, together with the impact of influenza infection on the BALF nucleotide profile, and the role of ERK MAP kinase in inducing nucleotide synthesis after influenza infection. 2) To determine the role of CD73 in influenza pathogenesis, effects of influenza infection on respiratory epithelial cell CD73 expression and activity will be measured; the impact of pharmacologic blockade of CD73 on influenza pathogenesis in C57BL/6 mice will be determined; and the outcome of influenza infection in CD73-knockout mice will be investigated. 3) To determine the role of adenosine/A1-AdoR receptor signaling in influenza pathogenesis, the effect of influenza-induced NF-kB activation on A1-AdoR expression in ATII cells and neutrophils will be measured; the impact of pharmacologic A1-AdoR blockade on influenza pathogenesis in C57BL/6 mice will be determined; and the outcome of influenza infection in A1-AdoR-/- knockout mice, or in A1- AdoR-/- bone marrow-chimeric mice will be investigated. Completion of these objectives will lead to increased understanding of the role of adenosine in the pathogenesis of influenza-associated lung injury, and permit determination of its potential as a target for therapeutic intervention to combat influenza-induced lung damage. PUBLIC HEALTH RELEVANCE: Influenza A virus causes a highly contagious acute respiratory disease. Seasonal influenza epidemics cause >300,000 deaths/yr worldwide, while pandemics elicited devastating loss of life in the 20th century, and may do so in the 21st. The proposed studies are aimed at improving our understanding of the mechanisms underlying lung dysfunction in influenza, and identifying a novel potential target for influenza therapy.

描述（由申请人提供）：甲型流感病毒引起高度传染性的急性呼吸道疾病。季节性流感流行每年在全球造成超过 30 万人死亡，而大流行在 20 世纪甚至在 21 世纪都造成了毁灭性的生命损失。然而，目前，流感或任何其他肺部病毒感染导致肺功能障碍和损伤的基本机制尚不清楚。 BALB/c 小鼠的呼吸道合胞病毒 (RSV) 和流感病毒感染都会诱导通道介导的核苷酸 UTP 和 ATP 释放到支气管肺泡内壁液 (BALF) 中，并且在这两种感染中，升高的 BALF UTP 会导致肺水肿和低氧血症的发生。 BALB/c 小鼠中的 RSV 感染仅引起轻微疾病，而流感会引起更严重的疾病，具有急性肺损伤的许多病理生理学特征，这在人类致命性大流行或流行性流感中也可见。重要的是，感染流感（而非 RSV）后，BALF 中 ATP 释放增加，同时 ATP 降解产物腺苷对 A1 亚型腺苷受体 (AdoR) 的激活增加。腺苷对中性粒细胞A1-AdoR的刺激在急性肺损伤的引发和促进中起重要作用。该提议的中心假设是，流感刺激受感染的 ATII 细胞从头合成和释放 ATP，释放的 ATP 以加速速率代谢为腺苷（由于核酸外切酶 CD73 活性增加），并且通过其对 A1-AdoR 的影响，BALF 中的腺苷在流感诱导的肺损伤中发挥着关键作用。该假设将在 3 个具体目标中进行测试：1) 为了确定流感感染对肺核苷酸代谢的暂时影响，将测量 MLE-12 细胞和纯化的肺泡 II 型 (ATII) 细胞以及来自 C57Bl/6 或 SP-C/GFP 小鼠的 FACS 分选的 ATII 细胞流感感染后核苷酸代谢的变化，以及 流感感染对 BALF 核苷酸谱的影响，以及 ERK MAP 激酶在流感感染后诱导核苷酸合成中的作用。 2) 为了确定CD73在流感发病机制中的作用，将测量流感感染对呼吸道上皮细胞CD73表达和活性的影响；将确定药物阻断 CD73 对 C57BL/6 小鼠流感发病机制的影响；并将研究 CD73 敲除小鼠的流感感染结果。 3) 为了确定腺苷/A1-AdoR受体信号传导在流感发病机制中的作用，将测量流感诱导的NF-kB激活对ATII细胞和中性粒细胞中A1-AdoR表达的影响；将确定药理 A1-AdoR 阻断对 C57BL/6 小鼠流感发病机制的影响；并将研究 A1-AdoR-/- 敲除小鼠或 A1-AdoR-/- 骨髓嵌合小鼠的流感感染结果。这些目标的完成将加深对腺苷在流感相关肺损伤发病机制中的作用的了解，并确定其作为对抗流感引起的肺损伤的治疗干预目标的潜力。 公共卫生相关性：甲型流感病毒会引起高度传染性的急性呼吸道疾病。季节性流感流行每年在全球造成超过 30 万人死亡，而大流行在 20 世纪甚至在 21 世纪都造成了毁灭性的生命损失。拟议的研究旨在提高我们对流感肺功能障碍机制的理解，并确定流感治疗的新潜在靶点。