The role of CFTR during macrophage-mediated killing of bacteria

项目总结 为什么囊性纤维化(CF)患者继续遭受慢性细菌感染 改善CF跨膜电导调节器(CFTR)功能的新药没有 为人所知。长期目标是开发调节宿主免疫反应的疗法。 用于缓解慢性炎症和慢性感染。这项建议的目的是 明确CFTR如何调节巨噬细胞的功能。这项建议背后的理由是 我们先前的工作表明，巨噬细胞是导致慢性阻塞性肺疾病患者能力低下的重要因素。 Cf通过失败的NADPH氧化酶(NOX)组装清除细菌感染，并减少 自噬。中心假说是人MΦ功能cftr缺失S抑制一氧化氮合酶 组装和随后的ROS介导的自噬，独立于CFTR突变类别，但是 由特定的机会主义细菌恶化。此外，我们预计CFTR的一个关键门槛 需要功能来逆转NOX组装/自噬缺陷，并且可以重新建立 通过CFTR调节器与替代CFTR修复剂(如半胱胺或 我们的新型自噬刺激器AR-13。核心假说将通过追查来检验 三个具体目标：1)确定CFtr调节M-Φ氮氧化物组装的机制；2) 确定特定病原菌如何差异调节M-ΦROS的产生；3)确定 新的治疗方法改变MΦ氮氧化物/自噬轴的程度。我们会 利用遗传学和药理学技术的创新组合来实现这些目标 人类巨噬细胞。这项拟议的研究具有重要意义，因为对 巨噬细胞功能是如何调节的将允许新的抗生素-和CFTR突变- 感染的不可知性治疗方法。它还具有重要意义，因为它将决定 特定的病原体独立地导致巨噬细胞介导的细菌杀灭的缺陷。 这项工作的预期结果将建立一个机制框架，使我们能够 靶向并纠正有缺陷的CFMΦ介导的细菌杀灭。最终，我们将把这个翻译成 将新知识转化为新的治疗范式，使用创新的宿主导向疗法 抗击细菌感染。

PROJECT SUMMARY Why patients with cystic fibrosis (CF) continue to suffer from chronic bacterial infections despite new medications that improve CF transmembrane conductance regulator (CFTR) function is not known. The long-term goal is to develop therapeutics that modulate host immune responses in CF patients to mitigate chronic infection and inflammation. The objective of this proposal is to define how CFTR regulates macrophage function. The rationale underlying this proposal is that our prior work demonstrates that CF macrophages are integral to the inability of patients with CF to clear bacterial infections through failed NADPH oxidase (NOX) assembly and reduced autophagy. The central hypothesis is that loss of functional CFTR in human MΦs inhibits NOX assembly and subsequent ROS-mediated autophagy, independent of CFTR mutation class, but worsened by specific opportunistic bacteria. Further, we expect that a critical threshold of CFTR function is needed to reverse the NOX assembly/autophagy deficits and can be re-established by CFTR modulators combined with alternative CFTR restoration agents such as cysteamine or our novel autophagy stimulator, AR-13. The central hypothesis will be tested by pursuing three specific aims: 1) Define the mechanism by which CFTR regulates MΦ NOX assembly; 2) Determine how CF specific pathogens differentially regulate MΦ ROS production; 3) Determine the extent to which novel therapeutic approaches alter the MΦ NOX/autophagy axis. We will pursue these aims using an innovative combination of genetic and pharmacologic techniques in human macrophages. The proposed research is significant because a precise understanding of how CF macrophage function is regulated would allow novel antibiotic- and CFTR mutation- agnostic treatment approaches to infection. It is also significant because it will determine if specific pathogens independently contribute to deficits in macrophage-mediated bacterial killing. The expected outcome of this work will establish a mechanistic framework to enable us to target and correct defective CF MΦ-mediated bacterial killing. Ultimately, we will translate this new knowledge into a new treatment paradigm that uses innovative host-directed therapies to combat bacterial infections.