Inducible Macrophage Specific Gene Expression in Diseases

疾病中诱导型巨噬细胞特异性基因表达

• 批准号: 7832227
• 负责人: Jeanine M D'Armiento
• 金额: $ 47.64万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7832227
• 关键词: Acute Lung Injury; Affect; Animal Model; ApoE knockout mouse; Apolipoprotein E; Apoptosis; Arterial Fatty Streak; Arthritis; Asthma; Atherosclerosis; Chronic; Chronic Obstructive Airway Disease; Cleaved cell; Development; Disease; Ensure; Exhibits; Foam Cells; Gene Expression; Generations; Genes; Goals; Heart; Human; Inflammation; Inflammatory; Injury; Investigation; Knock-out; Knockout Mice; Laboratories; Laboratory Study; Lesion; Lung; Modeling; Mus; Pathogenesis; Play; Reporter; Research Personnel; Role; Site; Specificity; Staging; System; Tamoxifen; Tissue Inhibitor of Metalloproteinase-3; Tissues; Transgenes; Transgenic Animals; Transgenic Mice; Vascular Diseases; angiogenesis; cigarette smoking; gene function; interest; knockout gene; lung injury; macrophage; migration; monocyte; mouse model; prevent; promoter; public health relevance; recombinase; scavenger receptor; tool

DESCRIPTION (provided by applicant): Develop transgenic animal models that are informative for understanding chronic inflammation in humans. Macrophages play a key role in the pathogenesis of multiple chronic inflammatory diseases. Studies using knockout mice have allowed investigators to examine the important role of macrophage-specific genes. However, these studies are limited to genes expressed in multiple tissues, as macrophage-specific knockout mice are not available. Since the loss of a gene may prevent the differentiation or migration of the monocyte prior to its actual arrival at the site of injury, the generation of an inducible, macrophage-specific knockout is preferable to study chronic inflammatory diseases. Our laboratory has developed a new transgenic mouse model targeting Cre recombinase expression in tissue macrophage, using the scavenger receptor A promoter (SRA-Cre). After its induction with tamoxifen, Cre will cleave any "floxed" gene in macrophages. Our model will also allow to knockout genes at any stage of the disease. The main goal of our proposal is to characterize this new conditional mouse model, using three aims. In the first aim, SRA-Cre mice will be crossed into a reporter line, and specificity and inducibility of Cre expression will be assessed in three models of lung injury (cigarette smoke exposure, asthma, and acute lung injury). In the second aim, SRA-Cre mice will be crossed into the apolipoprotein-E (Apoe) knockout background. Atherosclerotic plaques will be analyzed for expression of Cre in lesion macrophages and foam cells. This macrophage-specific Cre- expressing Apoe knockout model will be made available to laboratories studying the function of macrophages in atherosclerosis. In a third aim, we will generate of a conditional, macrophage- specific Tissue Inhibitor of Metalloproteinase-3 (Timp-3) knockout model. TIMP-3 is expressed in macrophage and plays an important role in inflammatory diseases. However, global Timp-3 knockout mice exhibited developmental abnormalities in the lung and the heart. We have generated a floxed Timp-3 model, which will be crossed into the SRA-Cre background. Utilizing this model, we will determine how the absence of TIMP-3 in macrophages affects lung injury due to cigarette smoke. Our new conditional, macrophage-specific targeting model will be a highly valuable tool, allowing researchers to precisely assess the roles and functions of genes expressed by macrophages during chronic inflammatory diseases. PUBLIC HEALTH RELEVANCE: Macrophages play a key role in the pathogenesis of multiple chronic inflammatory diseases. In this proposal, we will characterize a newly developed conditional, macrophage-specific mouse model, where Cre recombinase is regulated by the scavenger receptor A promoter. Our model will allow to knockout genes of interest only in macrophages and at any stage of the disease. The major goal of our proposal is to generate a mouse model that will allow researchers to specifically and conditionally knockout genes of interest in macrophages during chronic inflammatory diseases. This model will be a highly valuable model, allowing researchers to precisely assess the roles and functions of genes expressed by macrophages during chronic inflammatory diseases. In addition, we propose to cross this model into the atherosclerosis-prone Apoe knockout model, to allow examination of macrophage genes in this important vascular disease. We will also examine the consequences of the absence of macrophage tissue inhibitor of metalloproteinases-3 in lung injury.

描述（由申请人提供）：开发转基因动物模型，为了解人类慢性炎症提供信息。巨噬细胞在多种慢性炎症性疾病的发病机制中发挥着关键作用。使用基因敲除小鼠的研究使研究人员能够检查巨噬细胞特异性基因的重要作用。然而，这些研究仅限于在多个组织中表达的基因，因为还没有巨噬细胞特异性敲除小鼠。由于基因的丢失可能会阻止单核细胞在实际到达损伤部位之前的分化或迁移，因此产生可诱导的巨噬细胞特异性敲除对于研究慢性炎症性疾病来说是优选的。我们的实验室使用清道夫受体A启动子（SRA-Cre）开发了一种针对组织巨噬细胞中Cre重组酶表达的新型转基因小鼠模型。在用他莫昔芬诱导后，Cre 将裂解巨噬细胞中的任何“floxed”基因。我们的模型还允许在疾病的任何阶段敲除基因。我们提案的主要目标是使用三个目标来表征这种新的条件小鼠模型。第一个目标是，将 SRA-Cre 小鼠与报告系杂交，并在三种肺损伤模型（香烟烟雾暴露、哮喘和急性肺损伤）中评估 Cre 表达的特异性和诱导性。在第二个目标中，SRA-Cre 小鼠将进入载脂蛋白 E (Apoe) 敲除背景。将分析动脉粥样硬化斑块病变巨噬细胞和泡沫细胞中 Cre 的表达。这种巨噬细胞特异性 Cre 表达 Apoe 敲除模型将提供给研究巨噬细胞在动脉粥样硬化中功能的实验室。第三个目标是，我们将生成条件性巨噬细胞特异性金属蛋白酶-3 组织抑制剂 (Timp-3) 敲除模型。 TIMP-3在巨噬细胞中表达，在炎症性疾病中发挥重要作用。然而，全球 Timp-3 基因敲除小鼠表现出肺和心脏发育异常。我们生成了 floxed Timp-3 模型，该模型将交叉到 SRA-Cre 背景中。利用该模型，我们将确定巨噬细胞中 TIMP-3 的缺失如何影响香烟烟雾引起的肺损伤。我们新的条件性巨噬细胞特异性靶向模型将是一个非常有价值的工具，使研究人员能够精确评估慢性炎症性疾病期间巨噬细胞表达的基因的作用和功能。 公共卫生相关性：巨噬细胞在多种慢性炎症性疾病的发病机制中发挥着关键作用。在本提案中，我们将描述一种新开发的条件性巨噬细胞特异性小鼠模型，其中 Cre 重组酶受到清道夫受体 A 启动子的调节。我们的模型将允许仅在巨噬细胞和疾病的任何阶段敲除感兴趣的基因。我们提案的主要目标是生成一种小鼠模型，使研究人员能够在慢性炎症性疾病期间特异性地、有条件地敲除巨噬细胞中感兴趣的基因。该模型将是一个非常有价值的模型，使研究人员能够精确评估巨噬细胞表达的基因在慢性炎症疾病期间的作用和功能。此外，我们建议将该模型与易发生动脉粥样硬化的 Apoe 敲除模型相结合，以便检查这种重要血管疾病中的巨噬细胞基因。我们还将研究肺损伤中缺少金属蛋白酶-3 巨噬细胞组织抑制剂的后果。