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：
8241483
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/8241483
关键词：
Alleles Arginine Basic Science Biology Bleomycin CXCL10 gene CXCL11 gene CXCL9 gene CXCR3 gene Cell Proliferation Cells Collagen Data Development Drug Delivery Systems 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. PUBLIC HEALTH RELEVANCE: The project focuses on the role that macrophage phenotypes, particularly Alternatively Activated Macrophages have in fibrotic lung disease. This information could have important implications to our understanding of how macrophages could modulate the fibrotic response in humans. It could suggest that altering the function of macrophages and particularly arginase-1 would be a beneficial drug target for patients with pulmonary fibrosis.

描述（由申请人提供）：本提案研究的目标是提供支持和持续指导，以促进申请人发展成为间质性肺疾病和肺免疫学领域的独立科学研究人员。这将实现通过旨在获得扎实的基础科学背景的课程，包括实验室实践经验、定期的教学、频繁的演讲以及成功的医师科学家的密切指导。该提案将为申请人提供至少 75% 的受保护研究时间。利用该研究计划作为平台，申请人将获得许多基础科学技术的专业知识，包括：小鼠纤维化肺病的分析、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充满活力的研究小组，该小组在整个大学环境中发挥作用，促进前沿研究。研究项目：纤维化肺病是导致发病和死亡的重要原因，且缺乏有效的治疗方法。调节持续性肺纤维化的机制尚不完全清楚。我们的实验室最近描述了一种针对非感染性肺损伤的严重进行性纤维化模型。 CXCR3 是 IFNg 诱导型趋化因子 CXCL9 (Mig)、CXCL10 (IP-10) 和 CXCL11 (ITAC) 的受体。这些趋化因子被认为是 Th1 免疫的重要介质。与野生型对照小鼠相比，CXCR3 缺失小鼠在气管内博来霉素治疗后出现严重的进行性纤维化。微阵列分析已确定在缺乏 CXCR3 的情况下表达的与 Th2 免疫相关的大部分基因。此外，我们还鉴定了许多与替代激活的巨噬细胞表型相关的基因。这些包括甘露糖受体、YM1 和清道夫受体。与替代激活巨噬细胞表型相关的最显着的基因是精氨酸酶 1 的 36 倍诱导。虽然替代性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。 Arginase-1 由交替激活的巨噬细胞和成纤维细胞产生，以响应非感染性肺损伤。它具有直接介导纤维发生的潜力。 Arginase-1 负责将 L-精氨酸转化为脯氨酸和多胺。多胺参与细胞增殖，而脯氨酸是胶原蛋白的直接前体。巨噬细胞和成纤维细胞表达的精氨酸酶 1 在进行性纤维化中的作用尚未被探索。根据我们的初步数据，我们假设巨噬细胞和成纤维细胞中的精氨酸酶-1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。我们将在以下具体目标中检验这一假设：目标 1：使用区分招募群体和常驻群体的新型流式细胞术方法，定义非感染性肺损伤后 CXCR3 缺失小鼠招募到肺部的特定巨噬细胞表型。目标 2：使用 floxed arg-1 等位基因与靶向巨噬细胞或成纤维细胞的 cre 重组酶菌株杂交，确定非感染性损伤后巨噬细胞和成纤维细胞中精氨酸酶 1 的靶向删除的影响。总的来说，这种方法将使我们能够剖析巨噬细胞和成纤维细胞产生的精氨酸酶-1在进行性肺纤维化病理学中的作用。公共健康相关性：该项目重点关注巨噬细胞表型，特别是替代性激活巨噬细胞在纤维化肺部疾病中的作用。这些信息可能对我们理解巨噬细胞如何调节人类纤维化反应具有重要意义。这可能表明，改变巨噬细胞的功能，尤其是精氨酸酶-1，将成为肺纤维化患者有益的药物靶点。