Regulation of Lung Fibrosis by Alternatively Activated Macrophages and Arginase-1

选择性激活巨噬细胞和精氨酸酶 1 对肺纤维化的调节

基本信息

批准号：
8460814
负责人：
Robert Matthew Tighe
金额：
$ 9.06万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-01 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8460814
关键词：
Alleles Arginine Basic Science Biology Bleomycin CXCL10 gene CXCL11 gene CXCL9 gene CXCR3 gene Cell Proliferation Cells Collagen Data Development Drug Targeting Educational Curriculum Educational process of instructing Environment Enzymes Fibroblasts Fibrosis Fostering Generations Genes Goals Hand Human Immune Immunity Immunology Injury Interstitial Lung Diseases Knockout Mice Laboratories Lung Lung diseases Macrophage Activation Mediating Mediator of activation protein Mentors Mentorship Microarray Analysis Modeling Morbidity - disease rate Mus Patients Phenotype Physicians Play Polyamines Population Process Production Proline Pulmonary Fibrosis Recruitment Activity Regulation Relative (related person)Research Research Personnel Research Project Grants Research Proposals Role Scientist Solid Source Techniques Testing Time Tissues Universities arginase base career development cell type chemokine design effective therapy experience fibrogenesis lung development lung injury macrophage mannose receptor mortality novel novel strategies receptor recombinase response scavenger receptor

项目摘要

DESCRIPTION (provided by applicant): The goal of this proposal research is to provide support and continued mentorship to facilitate the applicant's development into an independent scientific investigator in interstitial lung disease and lung immunology. This will be accomplished through a curriculum designed to obtain a solid basic science background involving hands on laboratory experience, regular didactic teaching, frequent presentations, and close mentoring by successful physician-scientists. The proposal will allow at least 75% of protected research time for the applicant. Using the research proposal as a platform, the applicant will gain expertise in number of fundamental scientific techniques including: the analysis of murine fibrotic lung disease, the generation of cell specific knockout mice and the assessment of macrophage and fibroblast function. This effort will involve a dynamic research group which functions in an overal university environment fostering cutting edge research. Research Project: Fibrotic lung diseases are a significant cause of morbidity and mortality for which effective therapies are lacking. The mechanisms that regulate unremitting pulmonary fibrosis are incompletely understood. Our laboratory has recently described a model of severe and progressive fibrosis in response to non-infectious lung injury. CXCR3 is the receptor for the IFNg-inducible chemokines CXCL9 (Mig), CXCL10 (IP-10) and CXCL11 (ITAC). These chemokines have been suggested to be important mediators of Th1 immunity. CXCR3 null mice develop severe and progressive fibrosis following treatment with intratracheal bleomycin relative to wild type control mice. Microarray analysis has identified a preponderance of genes associated with Th2 immunity expressed in the absence of CXCR3. In addition, we identified a number of genes that have been associated with the alternatively activated macrophage phenotype. These include Mannose Receptor, YM1 and Scavenger Receptor. The most prominent of the genes associated with an alternatively activated macrophage phenotype was a 36-fold induction in arginase-1. While alternatively activated macrophages have been implicated in mediating immunologically mediated tissue fibrosis, their roles in non-immune regulated processes are less clear. Arginase-1 is produced both by alternatively activated macrophages and fibroblasts in response to non-infectious lung injury. It has the potential to directly mediate fibrogenesis. Arginase-1 is responsible for the conversion of L-arginine to proline and polyamines. Polyamines are involved in cell proliferation while proline is a direct precursor of collagen. The role of arginase-1 expression by macrophages and fibroblasts in progressive fibrosis has not been explored. Based on our preliminary data, we hypothesize that arginase-1 expression in macrophages and fibroblasts is critical to the development of fibrosis after non-infectious fibrotic lung injury. We will test thi hypothesis in the following specific aims: AIM 1: Define the specific macrophage phenotypes recruited to the lung in CXCR3 null mice after non-infectious lung injury using a novel flow cytometric approach that distinguishes recruited and resident populations. AIM2: Determine the effects of targeted deletion of arginase-1 in macrophages and fibroblasts after non-infectious injury using a floxed arg-1 allele crossed with either a cre-recombinase strain that targets macrophages or fibroblasts. Collectively, this approach will allow us to dissect the role of arginase-1 production by macrophages and fibroblasts in the pathobiology of progressive pulmonary fibrosis.

描述（由申请人提供）：该提案研究的目的是提供支持和持续指导，以促进申请人的发展成为间质性肺部疾病和肺部免疫学领域的独立科学研究者。这将完成通过旨在获得涉及实验室经验，常规教学，频繁演讲以及成功的医师科学家的良好基础科学背景的课程，涉及实验室的经验，经常的演讲和密切的指导。该提案将为申请人至少允许75％的受保护研究时间。将研究建议作为平台，申请人将获得有关基本科学技术数量的专业知识，包括：分析鼠纤维化肺部疾病，细胞特异性敲除小鼠的产生以及巨噬细胞和成纤维细胞功能的评估。这项工作将涉及一个动态的研究小组，该小组在促进尖端研究的大型大学环境中起作用。研究项目：纤维化肺部疾病是缺乏有效疗法的发病率和死亡率的重要原因。调节无肺纤维化的机制尚不完全理解。我们的实验室最近描述了一种针对非感染肺损伤的严重和进行性纤维化的模型。 CXCR3是IFNG诱导趋化因子CXCL9（MIG），CXCL10（IP-10）和CXCL11（ITAC）的受体。这些趋化因子被认为是Th1免疫力的重要介质。与野生型对照小鼠相对于气管内博来霉素治疗后，CXCR3无效小鼠在治疗后会出现严重的进行性纤维化。微阵列分析已经确定了在没有CXCR3的情况下表达的与Th2免疫相关的基因的优势。此外，我们确定了与替代激活的巨噬细胞表型相关的许多基因。这些包括甘露糖受体，YM1和清道夫受体。与替代激活的巨噬细胞表型相关的基因中最突出的基因是精氨酸酶-1的36倍诱导。否则，激活的巨噬细胞与介导免疫介导的组织纤维化有关，但它们在非免疫调节过程中的作用尚不清楚。精氨酸酶-1既是通过激活的巨噬细胞和成纤维细胞响应非感染性肺损伤产生的。它具有直接介导纤维发生的潜力。精氨酸酶1负责L-精氨酸向脯氨酸和多胺的转化。多胺参与细胞增殖，而脯氨酸是胶原蛋白的直接前体。尚未探索巨噬细胞和成纤维细胞在进行性纤维化中表达精氨酸酶-1的作用。根据我们的初步数据，我们假设巨噬细胞和成纤维细胞中精氨酸酶-1的表达对于非感染性纤维化肺损伤后纤维化的发展至关重要。我们将在以下特定目的中检验该假设：目标1：使用新型的流式细胞仪方法区分招募和居民种群，定义了非感染肺损伤后CXCR3无效小鼠肺部募集的特定巨噬细胞表型。 AIM2：使用Floxed Arg-1等位基因与靶向巨噬细胞或成纤维细胞的CRE-复异酶菌株交叉，确定非感染损伤后的精氨酸1缺失在巨噬细胞和成纤维细胞中的影响。总的来说，这种方法将使我们能够通过巨噬细胞和成纤维细胞在进行性肺纤维化病理生物学中剖析精氨酸酶-1产生的作用。