Role of GM-CSF in Myeloid Cell Function and Innate Immunity

GM-CSF 在骨髓细胞功能和先天免疫中的作用

基本信息

批准号：
8108866
负责人：
Bruce C Trapnell
金额：
$ 38.22万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8108866
关键词：
Address Affect Agonist Alveolar Macrophages Alveolus Asthma Autoantibodies Autoimmune Process Biological Biological Assay Biological Markers Bone Marrow Bronchoalveolar Lavage CD34 gene CSF2RA gene CSF2RB gene Cell Line Cell physiology Cells Chest Child Clinical Disease Event Excretory function Exposure to FDA approved Gene Transfer Genes Granulocyte-Macrophage Colony-Stimulating Factor Growth Hematopoietic stem cells Homeostasis Host Defense Human Immune System Diseases In Vitro Infection Inflammatory Inherited Irrigation Laboratories Lead Lipids Lung Macaca fascicularis Measures Mediating Methods Modeling Mus Mutation Myelogenous Myeloid Cells Natural History Natural Immunity Oral Outcome Measure Pathogenesis Patients Percussion Peroxisome Proliferator-Activated Receptors Pioglitazone Primates Procedures Pulmonary Alveolar Proteinosis Regulation Reporting Resolution Respiratory Failure Respiratory Insufficiency Rheumatoid Arthritis Role Saline Seminal Serum Signal Transduction Syndrome Testing Therapeutic Toxic effect Transcriptional Regulation Transplantation base cytokine immune function in vivo insight lentiviral-mediated lipid metabolism macrophage man mortality neutrophil novel novel diagnostics programs proto-oncogene protein Spi-1 receptor restoration surfactant translational clinical trial

项目摘要

DESCRIPTION (provided by applicant): Pulmonary alveolar proteinosis (PAP) is a syndrome characterized by accumulation of surfactant in alveolar macrophages (AMs) and alveoli resulting in respiratory failure and increased mortality from infection. For nearly 4 decades, the only available therapy was whole lung lavage, a highly invasive procedure performed at few centers in which one lung is mechanically ventilated while the other is repeatedly filled with saline and the chest is percussed vigorously to physically remove surfactant. No advances in pharmacologic therapy occurred due to a lack of pathogenic insight until PAP was discovered in GM-CSF-/- mice, a finding that transformed our concepts of the biological role of GM-CSF and led to novel diagnostics and therapy for PAP. My laboratory has contributed significantly to our understanding that GM-CSF is critical for surfactant homeostasis, AM ontogeny, neutrophil and AM functions, and innate immunity, and that in ~90% of patients, PAP is caused by a high level of GM-CSF autoantibodies (GMAbs). Current evidence suggests GM-CSF regulates surfactant homeostasis via the transcription factors PU.1 and PPAR? by stimulating expression of the lipid transporter, ABCG1: all three are deficient in AMs in GM-CSF-deficient mice and PAP patients. Notwithstanding, questions remain regarding the 1) natural history of PAP, 2) mechanism by which loss of GM-CSF signaling causes PAP, and 3) roles and relationship of PU.1 and PPAR3 in mechanisms by which GM-CSF regulates surfactant clearance and immune functions in AMs. We will use our novel primate model of autoimmune PAP, AM cell lines and PAP biomarkers; an existing murine model of hereditary PAP; and autoimmune and hereditary PAP patients to test our central hypothesis: PAP is caused by reduced GM-CSF?PU.1?PPAR3?ABCG1-dependent excretion of neutral lipids from AMs, which impairs their ability to clear surfactant. This hypothesis will be addressed in 3 specific aims focusing to GM-CSF regulation of myeloid cells. In Aim 1, we will determine the natural history of autoimmune PAP, critical threshold of GMAbs and their effects on myeloid immune functions in our primate model and PAP patients. In Aim 2, the roles of PU.1, PPAR?, and ABCG1 in hereditary PAP caused by CSF2RA or B mutations will be evaluated in vitro using lentiviral-mediated expression in macrophages from mice or humans with hereditary PAP, and in vivo by transplanting ABCG1-transduced bone marrow into CSF2RB-/- mice. In Aim 3, we will determine if GM-CSF regulates surfactant clearance and immune functions in AMs via the PU.1-dependent regulation of PPAR? using novel AM cell lines that do not spontaneously express PU.1, or that also respond to GM-CSF. The transcriptional program that GM-CSF regulates in AMs will be examined in vivo free of secondary effects of surfactant by using our primate model. We will determine if the PPAR? agonist pioglitazone restores AM surfactant clearance in vitro in cells from mice and humans with PAP and in vivo using CSF2RB-/- mice. Anticipated results have implications for PAP pathogenesis and therapy, surfactant homeostasis, and GMAb therapy of common inflammatory diseases. PUBLIC HEALTH RELEVANCE: GM-CSF is a cytokine regulator of alveolar macrophage function, surfactant homeostasis, and host defense. Disruption of GM-CSF signaling, due to GM-CSF autoantibodies or GM-CSF receptor mutations, causes pulmonary alveolar proteinosis (PAP), a syndrome of respiratory insufficiency due to surfactant accumulation in alveoli caused by reduced clearance by alveolar macrophages. In Aim 1, we will use a novel primate model of PAP and PAP patients to determine the natural history and level of GM-CSF autoantibodies causing PAP. In Aim 2, we will use gene transfer to determine the role PU.1, PPAR?, and ABCG1, in the pathogenesis of hereditary PAP and surfactant clearance by alveolar macrophages in vitro and in vivo (in mice). In Aim 3, we will determine if GM-CSF regulates PPAR3 in a PU.1-dependent manner and test whether pioglitazone, an FDA-approved PPAR? activator, is effective as therapy of hereditary PAP in vitro and in vivo (in mice). Anticipated results are relevant to surfactant homeostasis, pathogenesis and therapy of PAP, and the use of GM-CSF autoantibodies to treat common diseases like asthma and rheumatoid arthritis.

描述（由申请人提供）：肺泡蛋白沉积症（PAP）是一种综合征，其特征是表面活性剂在肺泡巨噬细胞（AM）和肺泡中积聚，导致呼吸衰竭和感染死亡率增加。近 4 年来，唯一可用的治疗方法是全肺灌洗，这是一种在少数中心进行的高度侵入性手术，其中一个肺进行机械通气，而另一个肺反复充满盐水，并剧烈敲击胸部以物理去除表面活性剂。由于缺乏对致病原因的了解，药物治疗一直没有取得进展，直到在 GM-CSF-/- 小鼠中发现 PAP，这一发现改变了我们对 GM-CSF 生物学作用的概念，并带来了新的 PAP 诊断和治疗方法。我的实验室对我们了解 GM-CSF 对于表面活性剂稳态、AM 个体发育、中性粒细胞和 AM 功能以及先天免疫至关重要的认识做出了重大贡献，并且在约 90% 的患者中，PAP 是由高水平的 GM-CSF 引起的自身抗体（GMAb）。目前的证据表明 GM-CSF 通过转录因子 PU.1 和 PPAR 调节表面活性剂稳态？通过刺激脂质转运蛋白 ABCG1 的表达：在 GM-CSF 缺陷小鼠和 PAP 患者中，这三种蛋白均缺乏 AM。尽管如此，关于 1) PAP 的自然史，2) GM-CSF 信号传导缺失导致 PAP 的机制，以及 3) PU.1 和 PPAR3 在 GM-CSF 调节表面活性剂清除和AM 中的免疫功能。我们将使用我们的新型灵长类动物模型，包括自身免疫性 PAP、AM 细胞系和 PAP 生物标志物；现有的遗传性 PAP 小鼠模型；和自身免疫性和遗传性 PAP 患者来测试我们的中心假设：PAP 是由 AM 中 GM-CSF？PU.1？PPAR3？ABCG1 依赖性中性脂质排泄减少引起的，这损害了它们清除表面活性剂的能力。这一假设将在 3 个具体目标中得到解决，重点是 GM-CSF 对骨髓细胞的调节。在目标 1 中，我们将确定自身免疫性 PAP 的自然史、GMAb 的临界阈值及其对我们的灵长类动物模型和 PAP 患者的骨髓免疫功能的影响。在目标 2 中，将使用来自患有遗传性 PAP 的小鼠或人类的巨噬细胞中的慢病毒介导的表达来体外评估 PU.1、PPAR? 和 ABCG1 在由 CSF2RA 或 B 突变引起的遗传性 PAP 中的作用，并通过移植来体内评估ABCG1 转导骨髓至 CSF2RB-/- 小鼠中。在目标 3 中，我们将确定 GM-CSF 是否通过 PU.1 依赖性 PPAR 调节来调节 AM 中的表面活性剂清除和免疫功能？使用不自发表达 PU.1 或也对 GM-CSF 有反应的新型 AM 细胞系。将使用我们的灵长类动物模型在体内检查 GM-CSF 在 AM 中调节的转录程序，而不受表面活性剂的二次影响。我们将确定是否有PPAR？激动剂吡格列酮可在体外恢复患有 PAP 的小鼠和人类细胞中的 AM 表面活性剂清除率，以及在体内使用 CSF2RB-/- 小鼠恢复 AM 表面活性剂清除率。预期结果对 PAP 发病机制和治疗、表面活性剂稳态以及常见炎症性疾病的 GMAb 治疗具有影响。公共卫生相关性：GM-CSF 是肺泡巨噬细胞功能、表面活性剂稳态和宿主防御的细胞因子调节剂。由于 GM-CSF 自身抗体或 GM-CSF 受体突变导致 GM-CSF 信号传导中断，会导致肺泡蛋白沉积症 (PAP)，这是一种呼吸功能不全的综合征，是由于肺泡巨噬细胞清除率降低导致肺泡内表面活性剂积聚所致。在目标 1 中，我们将使用 PAP 和 PAP 患者的新型灵长类动物模型来确定引起 PAP 的 GM-CSF 自身抗体的自然史和水平。在目标 2 中，我们将利用基因转移来确定 PU.1、PPAR? 和 ABCG1 在体外和体内（小鼠）遗传性 PAP 和肺泡巨噬细胞清除表面活性剂的发病机制中的作用。在目标 3 中，我们将确定 GM-CSF 是否以 PU.1 依赖性方式调节 PPAR3，并测试吡格列酮（FDA 批准的 PPAR）是否有效？激活剂，在体外和体内（小鼠）中可有效治疗遗传性 PAP。预期结果涉及表面活性剂稳态、PAP 的发病机制和治疗，以及使用 GM-CSF 自身抗体治疗哮喘和类风湿性关节炎等常见疾病。