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: 8819142
• 负责人: Bruce C Trapnell
• 金额: $ 36.97万
• 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 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8819142
• 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; Molecular; Mus; Mutation; Myelogenous; Myeloid Cells; Natural History; Natural Immunity; Oral; Outcome Measure; PPAR gamma; Pathogenesis; Patients; Percussion; Physiological; 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; public health relevance; 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 PPARγ 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→PPARγ→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, PPARγ, 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 PPARγ 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 PPARγ 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.

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通过刺激脂质转运蛋白ABCG 1的表达，通过转录因子PU.1和PPARγ调节表面活性物质稳态：在GM-CSF缺陷小鼠和PAP患者中，所有三种都是AM缺陷。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。我们将使用我们新的自身免疫性PAP灵长类动物模型、AM细胞系和PAP生物标志物;现有的遗传性PAP小鼠模型;以及自身免疫性和遗传性PAP患者来检验我们的中心假设：PAP是由AM中性脂质的GM-CSF→PU.1→PPARγ→ ABCG 1依赖性分泌减少引起的，这削弱了AM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，将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在体外评价PU. 1、PPARγ和ABCG 1在由CSF 2 RA或B突变引起的遗传性PAP中的作用，并通过将ABCG 1转导的骨髓移植到CSF 2 RB-/-小鼠体内评价。在目标3中，我们将使用不自发表达PU.1或也对GM-CSF有应答的新型AM细胞系，确定GM-CSF是否通过PU.1依赖性调节PPARγ来调节AM中的表面活性剂清除和免疫功能。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。我们将在PAP小鼠和人类细胞体外以及CSF 2 RB-/-小鼠体内确定PPARγ激动剂吡格列酮是否恢复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。