Role of GM-CSF in Alveolar Macrophage Self-Renewal

GM-CSF 在肺泡巨噬细胞自我更新中的作用

• 批准号: 10532222
• 负责人: Paritha Arumugam
• 金额: $ 54万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-01 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10532222
• 关键词: Adult; Agonist; Alveolar Macrophages; Biochemical; Biogenesis; CSF2RB gene; Cell Count; Cell Cycle Regulation; Cell Differentiation process; Cell Line; Cell Proliferation; Cell Survival; Cell Therapy; Cells; Child; Confocal Microscopy; Data; Development; Disease; Dyes; Enabling Factors; Evaluation; Fatty Acids; Feedback; Flow Cytometry; Foundations; Gene Expression Profiling; Genes; Genetic; Genus Hippocampus; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Growth; Half-Life; Health; Hematopoietic stem cells; Homeostasis; Hormones; Human; Impairment; Inherited; Interphase Cell; JAK2 gene; Knowledge; Leukemic Cell; Ligands; Lung; Lung Transplantation; Lung diseases; Macrophage; Macrophage Colony-Stimulating Factor; Maintenance; Measures; Mediating; Membrane Potentials; Metabolism; Methodology; Methods; Minor; Mission; Mitochondria; Molecular; Mus; Nature; Pathway interactions; Patients; Phase; Phospholipids; Play; Population; Population Sizes; Process; Production; Proliferating; Proteins; Public Health; Pulmonary Alveolar Proteinosis; Reactive Oxygen Species; Receptor Signaling; Regulation; Regulator Genes; Reporting; Research; Resolution; Respiration; Role; Signal Transduction; Source; Stains; Technology; Testing; Therapeutic; Transcriptional Regulation; Transplantation; United States National Institutes of Health; cell type; fatty acid oxidation; hematopoietic stem cell self-renewal; improved; in vivo; inhibitor; innovation; mitochondrial metabolism; monocyte; mouse model; novel; novel therapeutic intervention; novel therapeutics; oxidation; pharmacologic; progenitor; receptor; self renewing cell; self-renewal; surfactant; tool; transcription factor; transcriptome sequencing; transplantation therapy; vector

ABSTRACT Alveolar macrophages (AMs) are believed to be a self-renewing cell population without a requirement of replenishment from extra-pulmonary sources in healthy adult mice; however, the mechanism(s) involved and whether replenishment occurs by stimulating the proliferation of progenitors or mature AMs are not known. We reported that lung levels of granulocyte/macrophage-colony stimulating factor (GM-CSF) control long-term maintenance of macrophages trans- planted into the lungs (via a reciprocal feedback loop) as well as endogenous AMs. My preliminary data demonstrate that GM-CSF is a critical regulator of AM mitochondrial turnover, integrity and functions and is required for fatty acid oxidation- derived energy production, processes vital to cell proliferation. Prior studies in a leukemia cell line (TF-1) suggest the pleotropic effects of GM-CSF on macrophages may be mediated by biphasic, ligand concentration-dependent receptor signaling, i.e., low levels of GM-CSF promote survival and differentiation (but not proliferation) while high levels also stim- ulate proliferation. Our long-term goal is to determine mechanisms responsible for AM self-renewal. The objective here is to elucidate the mechanism by which GM-CSF regulation of mitochondrial homeostasis controls AM self-renewal. Cen- tral Hypothesis: AMs are maintained by homeostatic self-renewal driven by GM-CSF threshold-triggered/concentration- dependent, niche-limited proliferation of mature (long-lived) AMs (not progenitors). Rationale: This hypothesis was de- veloped based on my reported and preliminary data demonstrating GM-CSF deficiency results in reduced mitochondrial metabolism and integrity despite increased mitochondrial mass. Approach: I will utilize complementary genetic and phar- macologic tools, in vivo and ex vivo studies (with isolated AMs), and pathway-specific inhibitors to pursue two Specific Aims: 1) Ontogeny and transcriptional control of AM renewal, and 2) Role of GM-CSF regulated mitochondrial metabolism in AM renewal. The expected results will inform cellular and molecular mechanism(s) by which GM-CSF regulates AM population size and will lay the foundation for developing novel therapeutic strategies to modulate AM population size. The proposed research is innovative, in my opinion, because it challenges the previously widely-held concept of AMs as short-lived, non-dividing cells replenished from circulating monocytes (regulated by M-CSF) – and instead posits that AM population size is maintained by homeostatic self-renewal mediated by GM-CSF threshold-triggered/concentration-de- pendent, niche-limited proliferation of mature (long-lived) AMs. My novel preliminary data identified GM-CSF dependent regulation of mitochondrial homeostasis as a molecular mechanism for AM self-renewal. In addition, utilization of novel mouse models and novel methodologies will enable determination of how GM-CSF regulation of mitochondrial homeosta- sis regulates AM proliferation and will identify the regulatory genes and their downstream targets responsible for long- term maintenance of AMs. The proposed research is significant because it will advance our knowledge of a critical pulmo- nary hormone (GM-CSF) by informing the mechanism(s) by which GM-CSF regulates the long-term maintenance of endog- enous and transplanted AMs and inform us on the therapeutic mechanism of action of PMT, a novel cell therapy in devel- opment to treat patients with hereditary pulmonary alveolar proteinosis and potentially other lung diseases.

抽象的 据信肺泡巨噬细胞（AMS）是一个自我更新的细胞种群，无需补充 来自健康的成年小鼠的肺外来源；但是，涉及的机制以及是否复制 通过刺激祖细胞或成熟AM的增殖而发生。我们报告说肺水平 粒细胞/巨噬细胞 - 颜色刺激因子（GM-CSF）控制巨噬细胞的长期维持 种植到肺（通过相互反馈回路）和内源性AM中。我的初步数据表明 GM-CSF是AM线粒体周转，完整性和功能的关键调节剂，是脂肪酸氧化所必需的 得出的能源产生，对细胞增殖至关重要。在白血病细胞系（TF-1）中的先前研究表明 GM-CSF对巨噬细胞的多性作用可能是由双相，配体浓度依赖性受体介导的 信号传导，即GM-CSF的低水平促进生存和分化（但不是增殖），而高水平也刺激了 Ulate增殖。我们的长期目标是确定负责自我更新的机制。这里的目标 是为了阐明GM-CSF调节线粒体稳态控制的机制。 c TRAL假设：通过稳态自我更新驱动器通过GM-CSF阈值触发/浓度来维持AM 成熟（长寿命）AM（不是祖细胞）的依赖性，限制的限制增殖。理由：这一假设是 根据我的报告和初步数据，证明GM-CSF缺乏症会导致线粒体减少 代谢和诚信任务增加了线粒体质量。方法：我将利用完整的遗传和阶段 Macogogic工具，体内和离体研究（具有孤立的AM）和特定途径的抑制剂，可购买两种特定的特定 目的：1）AM更新的本体发育和转录控制，2）GM-CSF调节的线粒体代谢的作用 在Am Renewal中。预期的结果将为GM-CSF调节AM的细胞和分子机制提供信息 人口规模，将奠定基础，以制定新型的热策略来调节AM人口规模。 在我看来，拟议的研究具有创新性，因为它挑战了以前的AMS概念 短寿命的非分散细胞从循环单核细胞（由M-CSF调节）中取代，而是假定为AM 由GM-CSF阈值触发/浓度-DE-介导的稳态自我更新维持人口大小。 成熟（长寿命）AM的吊坠，限制的利基限制扩散。我的新型初步数据确定了GM-CSF依赖性 线粒体稳态的调节作为AM自我更新的分子机制。另外，新颖的利用 小鼠模型和新方法将确定GM-CSF如何调节线粒体稳态的调节 SIS调节AM增殖，并将确定调节基因及其下游靶标 AMS的定期维护。拟议的研究很重要，因为它将促进我们对关键的Pulmo-的了解 通过通知GM-CSF的机制，纳里激素（GM-CSF）调节了内og-的长期维持 由AM和移植AM并告知我们PMT的治疗机理，PMT是一种新型细胞疗法 治疗遗传性肺肺泡蛋白质和可能其他肺部疾病的患者的选择。