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

• 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
• 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

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）是一种以肺泡巨噬细胞（AM）和肺泡中表面活性剂蓄积为特征的综合征，导致呼吸衰竭和感染死亡率增加。近40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，而另一个肺反复填充盐水，胸部剧烈扩张以物理去除表面活性剂。由于缺乏对病原体的了解，药物治疗没有进展，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，并导致PAP的新诊断和治疗。我的实验室为我们理解GM-CSF对表面活性物质稳态、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自身抗体（GMAb）引起的。目前的证据表明GM-CSF通过转录因子PU.1和PPAR？通过刺激脂质转运蛋白ABCG 1的表达：在GM-CSF缺陷小鼠和PAP患者中，所有三种都是AM缺陷型。尽管如此，关于1）PAP的自然史，2）GM-CSF信号传导丢失引起PAP的机制，和3）PU 1和PPAR 3在GM-CSF调节AM中表面活性剂清除和免疫功能的机制中的作用和关系，仍然存在问题。我们将使用我们的新的自身免疫性PAP，AM细胞系和PAP生物标志物的灵长类动物模型;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是由减少GM-CSF？PU.1？PPAR3？ABCG 1依赖性的中性脂质从AM的排泄，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，重点是骨髓细胞的GM-CSF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，PU.1、PPAR？、和ABCG 1在由CSF 2 RA或B突变引起的遗传性PAP中的表达将在体外使用慢病毒介导的来自患有遗传性PAP的小鼠或人的巨噬细胞中的表达进行评估，并且在体内通过将ABCG 1转导的骨髓移植到CSF 2 RB-/-小鼠中进行评估。在目的3，我们将确定是否GM-CSF调节表面活性剂清除和免疫功能的AM通过PU。1依赖性调节的PPAR？使用不自发表达PU.1或也对GM-CSF应答的新AM细胞系。将通过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次级作用。我们将确定是否PPAR？激动剂吡格列酮在来自患有PAP的小鼠和人的细胞中体外恢复AM表面活性剂清除，并且在使用CSF 2 RB-/-小鼠的体内恢复AM表面活性剂清除。预期的结果对PAP的发病机制和治疗，表面活性物质的稳态，和常见的炎症性疾病的GMAb治疗的影响。 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受体突变导致的GM-CSF信号传导的破坏导致肺泡蛋白沉积症（PAP），这是一种由于肺泡巨噬细胞清除减少引起的肺泡中表面活性剂积聚而导致的呼吸功能不全综合征。在目标1中，我们将使用一种新的PAP和PAP患者的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，我们将使用基因转移来确定PU.1，PPAR？，和ABCG 1，在遗传性PAP和肺泡巨噬细胞在体外和体内（小鼠）的表面活性剂清除的发病机制。在目标3中，我们将确定GM-CSF是否以PU.1依赖的方式调节PPAR 3，并测试FDA批准的PPAR 3？活化剂，在体外和体内（小鼠）作为遗传性PAP的治疗有效。预期结果与PAP的表面活性物质稳态、发病机制和治疗以及GM-CSF自身抗体治疗常见疾病如哮喘和类风湿性关节炎有关。