The Role of Macrophage Metabolic Crosstalk in CF Chronic Lung Inflammation

项目摘要 囊性纤维化(CF)患者患有慢性感染和肺部炎症，导致支气管扩张 最终导致呼吸衰竭。尽管最近的进展，包括批准高效的CFTR 调节剂疗法(HEMT)，改善了慢性阻塞性肺疾病(PwCF)患者的整体生活质量 感染和炎症是发病的主要原因。尽管炎症的严重程度 已知在CF肺中反应增加，导致持续性炎症的机制是 未知。我们已经证明，肺巨噬细胞在CF的局部炎症反应中起关键作用。肺 巨噬细胞包括常驻的肺巨噬细胞，以及被招募到肺的肺巨噬细胞。 对炎性刺激的反应。我们的初步数据表明，CF募集了肺巨噬细胞 减少了Nrf2的表达，Nrf2是一种已知调节细胞代谢的转录因子 非CF型支气管扩张症患者和对照组，HEMT治疗后这种情况没有改善。我们还发现，CF 招募的肺巨噬细胞持续糖酵解和炎症，即使在HEMT的背景下，而 非慢性纤维支气管扩张症和健康招募的肺巨噬细胞可以过渡到炎症消退 表型。由于免疫细胞串扰可以由细胞外小泡(EVS)介导，我们研究了 驻留肺巨噬细胞EVS对肺巨噬细胞炎症反应的影响 发现滞留肺巨噬细胞的EV可诱导新招募的肺巨噬细胞的持续性炎症。 最后，我们有初步数据显示，预计会抑制Nrf2的miRNAs增加，而 肺巨噬细胞来源的EVS中的炎症分解脂质。因此，我们假设特定的 CF驻留LMEV内的miRNAs和脂质降低了招募LMS中的Nrf2水平，导致持久 糖酵解和未能转变为炎症消退表型。在目标1中，我们将检验假设 肺巨噬细胞在功能上存在重要的免疫代谢差异，这些差异是特异性的 Cf并在HEMT后持续。在这个目标中，我们将充分描述肺巨噬细胞亚群的特征 并将量化居民和居民在细胞代谢和炎症消退方面的差异 在PwCF、非CF支扩患者和健康受试者中招募肺巨噬细胞。在目标2中，我们将 验证由CFs驻留肺巨噬细胞释放的EV中的特定miRNAs影响 募集的肺巨噬细胞的炎症反应。在目标3中，我们将检验这样一个假设，即 肺泡巨噬细胞释放的EVS的含量可阻止炎症消退 募集的肺巨噬细胞的表型。拟议中的研究是独一无二的，因为它们涉及人类 HEMT前后的受试者，因此，我们的数据将直接与PwCF.另外，我们的研究 将为持续肺炎症的发病机制提供新的重要信息 使我们能够确定新疗法的靶点，以减少有害的CF肺部炎症并改善生活 和生活在CF患者身上的寿命。

Project Summary Patients with cystic fibrosis (CF) suffer from chronic infections and lung inflammation leading to bronchiectasis and, ultimately, respiratory failure. Although recent advances, including the approval of highly effective CFTR modulator therapy (HEMT), have improved the overall quality of life of people with CF (PwCF), chronic infection and inflammation are the primary cause of morbidity. Although the magnitude of the inflammatory response is known to be increased in the CF lung, the mechanisms underlying persistent inflammation are unknown. We have shown that lung macrophages are critical to the local inflammatory response in CF. Lung macrophages include resident lung macrophages, as well as lung macrophages that are recruited to the lung in response to inflammatory stimuli. Our preliminary data demonstrate that CF recruited lung macrophages have decreased expression of Nrf2, a transcription factor known to regulate cellular metabolism, compared to non-CF bronchiectasis and control subjects, and this did not improve with HEMT. We also found that CF recruited lung macrophages are persistently glycolytic and inflammatory, even in the setting of HEMT, while non-CF bronchiectasis and healthy recruited lung macrophages can transition to an inflammation resolving phenotype. As immune cell crosstalk can be mediated by extracellular vesicles (EVs), we investigated the impact of resident lung macrophage EVs on the inflammatory response of recruited lung macrophages and found that CF resident lung macrophage EVs induce persistent inflammation in recruited lung macrophages. Lastly, we have preliminary data showing increased miRNAs predicted to inhibit Nrf2 and reduced levels of inflammation resolving lipids in EVs from CF resident lung macrophages. Thus, we hypothesize that specific miRNAs and lipids within CF resident LM EVs reduce Nrf2 levels in recruited LMs, causing persistent glycolysis and failure to transition to an inflammation resolving phenotype. In Aim 1, we will test the hypothesis that there are functionally important immunometabolic differences in CF lung macrophages that are specific to CF and persist after HEMT. In this Aim, we will fully characterize subpopulations of lung macrophages in the CF lung and will quantify differences in cellular metabolism and inflammation resolution between resident and recruited lung macrophages in PwCF, non-CF bronchiectasis subjects, and healthy subjects. In Aim 2, we will test the hypothesis that specific miRNAs within EVs released by CF resident lung macrophages impact the inflammatory response of recruited lung macrophages. In Aim 3, we will test the hypothesis that the lipid content of EVs released by CF resident lung macrophages prevents the shift to an inflammation resolution phenotype in recruited lung macrophages. The proposed studies are unique because they involve human subjects before and after HEMT and thus, our data will be directly relevant to PwCF. In addition, our studies will provide new and essential information on the mechanisms of persistent lung inflammation in CF and will allow us to identify targets for novel therapies to reduce harmful CF lung inflammation and improve the lives and longevity of people living with CF.