Mouse Asthma

小鼠哮喘

• 批准号: 7476207
• 负责人: DANIEL H CONRAD
• 金额: $ 18.59万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7476207
• 关键词: Aerosols; Affect; Allergens; Allergic; Animals; Asthma; Breathing; Cells; Chemicals; Collaborations; Collection; Condition; Core Facility; Data Analyses; Development; Disease; Disease model; Effectiveness; Evaluation; Functional disorder; Genetic Variation; Histology; Human; Hypersensitivity; IDEC-152 Monoclonal Antibody; IgE; Infiltration; Inflammation; Intranasal Administration; Laboratories; Lead; Life; Lipids; Liquid substance; Lung; Maintenance; Measurement; Mediating; Membrane; Methodology; Methods; Modality; Modeling; Monitor; Mus; Operative Surgical Procedures; Organ; Ovalbumin; Pathology; Phenotype; Phosphotransferases; Preparation; Production; Protein Overexpression; RNA analysis; Relative (related person); Role; Route; SPHK1 enzyme; Small Interfering RNA; Sphingosine; Symptoms; System; Testing; Therapeutic Intervention; Toxic effect; Transgenic Organisms; aerosolized; chemokine; concept; cytokine; day; eosinophil; in vivo; inhibitor/antagonist; knock-down; knockout animal; mast cell; mouse model; novel; programs; protective effect; research study; response; sphingosine kinase; src-Family Kinases; transcription factor

The mouse asthma model core (scientific core C) will coordinate and perform the experiments in projects 2, 3 and 4 that involve the mouse asthma model. In Project 4, the Leader, Dr. Spiegel, has developed specific inhibitors for sphingosine-1-kinase and will use these in an inhalent model to get them directly to the lung. This largely avoids systemic toxicity and targets the affected organ directly. The concept being tested is whether inhibition of lung mast cells is sufficient to inhibit the asthma model. In addition, the program director will use siRNA to block, in vivo, in lung the synthesis of sphingosine kinase in order to determine whether this will inhibit asthma development. In Project 2, Dr. Conrad has demonstrated that overexperssion of CD23 inhibits IgE production and eosinophil recruitment to the lung. Experiments will be performed to determine if IgE is necessary for the eosinophil recruitment. In addition, the CD23 sheddase has been identified as ADAM 10 and siRNA knock-down experiments will be performed in collaboration with Dr. Spiegel to determine if blockade of ADAM10 in the lung, either by siRNA or specific inhibitors influences asthma development and symptoms. In Project #3, Dr. Ryan is examining the allergic phenotype of lyn knockout animals with the objective of determining the activities of lyn in the mast cell. The lyn"'" mice are known to develop severe Th2 disease including increased IgE and eosinophil responses. These activities, not surprisingly, lead to a severe asthma phenotype. In collaboration with Dr, Conrad, the lyn"'" animals will be crossed to CD23 transgenics to see if the Th2 disease is modulated. In addition, the activities of ADAM10 inhibitors either via a siRNA or chemical inhibitor methodology, introduced directly into the lung, will be tested for capacity to modulate the lyn"'" severe Th2 disease modality. In all of these studies, the core will provide the expertise to both perform and analyze data from the mouse asthma model. These include maintenance of mice for the 30 day period required for sensitization, aerosol challenge and ultimate sacrifice with BAL fluid collection and lung preparation for histology and RNA analysis.

小鼠哮喘模型核心（科学核心C）将协调和执行项目中的实验 图2、3和4涉及小鼠哮喘模型。在项目4中，领导者Spiegel博士开发了 鞘氨醇-1-激酶的特异性抑制剂，并将在吸入模型中使用这些药物， 肺这在很大程度上避免了全身毒性，并直接靶向受影响的器官。这个概念是 测试肺肥大细胞的抑制是否足以抑制哮喘模型。此外该 项目负责人将使用siRNA在体内阻断肺中鞘氨醇激酶的合成， 确定这是否会抑制哮喘的发展。在项目2中，康拉德博士已经证明， CD 23的过度表达抑制IgE产生和嗜酸性粒细胞向肺的募集。实验将 以确定IgE是否是嗜酸性粒细胞募集所必需的。此外，CD 23 脱落酶已被鉴定为ADAM 10，siRNA敲低实验将在 与Spiegel博士合作，以确定是否通过siRNA或特异性阻断肺中的ADAM 10， 抑制剂影响哮喘的发展和症状。在项目#3中，Ryan博士正在检查 Lyn林恩敲除动物的过敏表型，目的是确定林恩在 肥大细胞已知林恩+/-小鼠发展严重的Th 2疾病，包括增加的IgE， 嗜酸性粒细胞反应。毫不奇怪，这些活动导致严重的哮喘表型。在 与康拉德博士合作，将把林恩动物与CD 23转基因动物杂交，看看Th 2是否 疾病是被调制的。此外，通过siRNA或化学药物的ADAM 10抑制剂的活性 将测试直接引入肺中的抑制剂方法调节林恩的能力。 严重的Th 2疾病模式。在所有这些研究中，核心将提供专业知识， 并分析小鼠哮喘模型的数据。这些包括维持小鼠30天 致敏、气溶胶激发和最终处死（采集BAL液）所需的时间， 用于组织学和RNA分析的肺制备。