The pivotal role of macrophages in regulating pulmonary fibrosis

巨噬细胞在调节肺纤维化中的关键作用

• 批准号: 10133125
• 负责人: Megan N Ballinger
• 金额: $ 50.51万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10133125
• 关键词: Adoptive Transfer; Affect; Alveolar Macrophages; Animal Model; Biological Markers; Bleomycin; Bone Marrow; Bone Marrow Transplantation; CCL2 gene; Cell Communication; Cell Culture System; Cells; Coculture Techniques; Collagen; Complex; Data; Deposition; Development; Diagnosis; Disease; Disease Progression; Elasticity; Embryo; Environment; Enzymes; Fibroblasts; Fibrosis; Foundations; Gene Expression; Generations; Genes; Genetic Transcription; Goals; Health; Human; IRAK3 gene; In Vitro; Inflammation; Injury; Interleukin-13; Intervention; Knockout Mice; Knowledge; Lead; Life Expectancy; Lung; Lung Transplantation; Matrix Metalloproteinases; Measures; Modeling; Molecular Target; Mus; Myeloid Cells; Nature; Patients; Peripheral Blood Mononuclear Cell; Pharmacology; Phenotype; Play; Population; Production; Proliferating; Publishing; Pulmonary Fibrosis; Receptor Signaling; Resistance; Role; Sampling; Sensory; Signal Transduction; System; Testing; Tissues; Toll-like receptors; United States; Work; Yolk Sac; base; cell type; chemokine receptor; design; idiopathic pulmonary fibrosis; improved; inhibitor/antagonist; innovation; insight; macrophage; monocyte; mouse model; new therapeutic target; novel; novel marker; novel therapeutics; promoter; receptor; recruit; trafficking; transplant model; treatment strategy; uptake

PROJECT SUMMARY/ABSTRACT Idiopathic Pulmonary Fibrosis (IPF) is a progressive and fatal disorder of excessive collagen deposition by fibroblasts resulting in reduced lung elasticity and poor survival. Despite ongoing efforts and new pharmacologic agents, only limited efficacy has been achieved to delay disease progression. This likely reflects the complex nature of the disease including vital cell-cell interactions. Recently, we identified critical interactions between macrophages and fibroblasts which led to the development of pulmonary fibrosis. Our data demonstrate that interleukin-1 receptor associated kinase (IRAK)-M, a negative regulator of Toll-like receptor signaling, was elevated in macrophages from human IPF patients and murine macrophages after bleomycin-induced pulmonary fibrosis and lead to enhanced collagen expression by fibroblasts. Using our experimental murine model of fibrosis, we demonstrate that mice deficient in IRAK-M are protected from pulmonary fibrosis and this effect is dependent on IRAK-M expression in macrophages. The goal of this proposal is to investigate the mechanism by which macrophage expression of IRAK-M regulates the development of pulmonary fibrosis. In order to accomplish this, we propose two specific aims. In specific aim 1, we will investigate the role of IRAK-M in regulating monocyte trafficking to the lung during pulmonary fibrosis. Lung macrophage are either derived embryonically from the yolk-sac or are bone marrow-derived from circulating monocytes. We will interrogate the role of IRAK-M in regulating monocyte trafficking using adoptive transfer, competitive bone marrow transplantation model, and cell-specific IRAK-M knockout mice. In addition, we will investigate the role of IRAK-M in regulating expression of profibrotic genes and CCR2, the chemokine receptor involved in monocyte recruitment, using macrophages isolated from bleomycin challenged mice as well as primary macrophages from IPF patients and normal, donor controls. To show translational relevance, we will use our characterized human samples to generate a humanized IPF mouse model with cells from IPF patients. In specific aim 2, we will determine the role of IRAK-M in regulating macrophage function, specifically their ability to identify, take up and degrade collagen in the lung during pulmonary fibrosis. We will assess the role of IRAK-M in the uptake of collagen fragments after bleomycin challenge. Macrophage and monocyte populations from our murine models as well as cells isolated from IPF patients and normal donor controls will be flow sorted and we will measure the expression of IRAK-M, collagen uptake receptors, and collagen degradation enzyme expression and activity. Finally, we will establish a macrophage-fibroblast co-culture system to investigate the bidirectional interactions that these two cell types exert on each other. These studies will provide the foundation for the development of novel biomarkers of disease as well as new interventions based on specific molecular targets that should improve the human health of IPF patients.

项目总结/摘要 特发性肺纤维化（IPF）是一种进行性和致命的过度胶原沉积疾病， 成纤维细胞导致肺弹性降低和存活率差。尽管不断努力， 尽管已经使用了多种药物，但仅实现了有限的功效来延迟疾病进展。这可能 反映了疾病的复杂性质，包括重要的细胞-细胞相互作用。最近，我们发现了 巨噬细胞和成纤维细胞之间的相互作用，导致肺纤维化的发展。我们 数据表明，白细胞介素-1受体相关激酶（IRAK）-M，Toll样的负调节因子， 受体信号传导，在人IPF患者的巨噬细胞和小鼠巨噬细胞中升高， 博来霉素诱导的肺纤维化，并导致成纤维细胞胶原蛋白表达增强。使用我们 在实验性小鼠纤维化模型中，我们证明IRAK-M缺陷的小鼠在纤维化发生时受到保护， 肺纤维化，并且该作用依赖于巨噬细胞中的IRAK-M表达。这个目标 我们的建议是研究巨噬细胞表达IRAK-M调节细胞凋亡的机制。 肺纤维化的发展。为了实现这一目标，我们提出了两个具体目标。在具体目标1中， 我们将研究IRAK-M在肺纤维化过程中调节单核细胞运输到肺中的作用。 肺巨噬细胞要么来自卵黄囊的胚胎，要么来自骨髓 循环单核细胞我们将使用过继免疫荧光技术研究IRAK-M在调节单核细胞运输中的作用。 转移、竞争性骨髓移植模型和细胞特异性IRAK-M敲除小鼠。此外，本发明还提供了一种方法， 我们将研究IRAK-M在调节促纤维化基因和趋化因子CCR 2表达中的作用。 受体参与单核细胞募集，使用从博来霉素攻击的小鼠中分离的巨噬细胞作为 以及来自IPF患者和正常供体对照的原代巨噬细胞。为了显示翻译的相关性， 我们将使用我们表征的人类样品，用来自IPF的细胞产生人源化IPF小鼠模型， 患者在具体目标2中，我们将确定IRAK-M在调节巨噬细胞功能中的作用，具体地说， 它们在肺纤维化期间识别、摄取和降解肺中胶原蛋白的能力。我们将评估 IRAK-M在博来霉素激发后胶原片段摄取中的作用。巨噬细胞和单核细胞 来自我们的鼠模型的细胞群以及从IPF患者和正常供体对照分离的细胞将 进行流式分选，我们将测量IRAK-M、胶原摄取受体和胶原蛋白的表达。 降解酶的表达和活性。最后，我们将建立巨噬细胞-成纤维细胞共培养 系统来研究这两种细胞类型彼此施加的双向相互作用。这些研究 将为开发新的疾病生物标志物和新的干预措施提供基础 基于应改善IPF患者人类健康的特定分子靶点。