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）：肺肺泡蛋白质病（PAP）是一种综合征，其特征是表面活性剂在肺泡巨噬细胞（AMS）（AMS）和肺泡中的积累，导致呼吸衰竭和感染死亡率增加。近40年来，唯一可用的疗法是整个肺灌洗，这是在几个中心进行的高度侵入性手术，其中一种肺是机械通气的，另一种肺部被反复填充盐水，胸部被剧烈敲击以进行物理去除表面活性剂。由于缺乏致病性洞察力，直到在GM-CSF - / - 小鼠中发现PAP，这一发现改变了我们对GM-CSF的生物学作用的概念并导致PAP的新型诊断和治疗，因此没有任何进展。我的实验室为我们的理解做出了重大贡献，即GM-CSF对于表面活性剂稳态，AM的发育，中性粒细胞和AM功能以及先天免疫力至关重要，而在约90％的患者中，PAP是由高水平的GM GM-CSF自身抗体（GMABS）引起的。当前的证据表明，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的表达，ABCG1：在GM-CSF缺陷型小鼠和PAP患者中，这三个均缺乏AMS。尽管如此，关于1）PAP的自然历史的问题仍然存在，2）GM-CSF信号传导的损失会导致PAP，以及3）PU.1和PPAR3在AMS中调节表面活性剂清除和免疫功能的机制中的作用和关系。我们将使用自身免疫性PAP，AM细胞系和PAP生物标志物的新型灵长类动物模型；现有的遗传性帕普的鼠模型；和自身免疫性和遗传性PAP患者测试我们的中心假设：PAP是由GM-CSF？PU.1？PPAR3？pPAR3？aBCG1依赖性排泄中性脂质的排泄，这会损害其清除表面活性剂的能力。该假设将在3个特定目的中解决，重点是髓样细胞的GM-CSF调节。在AIM 1中，我们将确定自身免疫性PAP的自然历史，GMAB的临界阈值及其对我们的灵长类动物模型和PAP患者中对髓样免疫功能的影响。在AIM 2中，PU.1，PPAR？和ABCG1在由CSF2RA或B突变引起的遗传性PAP中的作用将在体外使用慢病毒介导的遗传噬细胞中的巨噬细胞中的表达进行评估，或者通过遗传性PAP的巨噬细胞中的巨噬细胞表达，并通过将ABCG1-Transplant Abcg1-Trans-trans-trans transpap trans transpap transpaps trans-transpap trance transpap trance transpap。在AIM 3中，我们将确定GM-CSF是否通过PPAR的pa.1依赖性调节AMS调节表面活性剂清除率和免疫功能？使用不会自发表达PU.1的新型AM细胞系，或者也对GM-CSF做出响应。 GM-CSF在AM中调节的转录程序将在体内使用我们的灵长类动物模型，不受表面活性剂的次要作用检查。我们将确定PPAR是否？激动剂吡格列酮使用CSF2RB - / - 小鼠在小鼠和人体的小鼠和体内的细胞中恢复体外表面活性剂清除率。预期的结果对PAP发病机理和治疗，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。 GM-CSF信号传导的破坏是由于GM-CSF自身抗体或GM-CSF受体突变引起的，导致肺肺泡蛋白质病（PAP），这是一种由于肺泡巨噬细胞降低引起的肺泡降低引起的肺泡综合症综合征。在AIM 1中，我们将使用新型的PAP患者的灵长类动物模型来确定引起PAP的GM-CSF自身抗体的自然病史和水平。在AIM 2中，我们将使用基因转移来确定PU.1，PPAR？和ABCG1的作用，在体外和体内肺泡巨噬细胞的遗传性PAP和表面活性剂清除率（在小鼠中）。在AIM 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Pioglitazone是否是FDA批准的PPAR？激活剂在体外和体内（在小鼠中）有效地作为遗传性子宫颈囊治疗。预期的结果与表面活性剂稳态，PAP的发病机理和治疗以及使用GM-CSF自身抗体治疗常见疾病（如哮喘和类风湿关节炎）有关。