The role of CFTR during macrophage-mediated killing of bacteria

项目摘要 为什么囊性纤维化（CF）患者继续遭受慢性细菌感染，尽管 改善CF跨膜传导调节因子（CFTR）功能的新药物 知道的长期目标是开发调节宿主免疫反应的疗法 以减轻慢性感染和炎症。这项建议的目的是 定义CFTR如何调节巨噬细胞功能。这项建议的基本理由是， 我们先前的工作表明CF巨噬细胞是导致患者不能正常生活的一个组成部分。 CF通过失败的NADPH氧化酶（NOX）组装清除细菌感染， 自噬中心假设是人M Φ中功能性CFTR的丧失抑制了NOX 组装和随后的ROS介导的自噬，独立于CFTR突变类别，但 会被特殊的机会性细菌破坏此外，我们预计CFTR的临界阈值 需要逆转NOX组装/自噬缺陷，并且可以重新建立 通过CFTR调节剂与替代的CFTR恢复剂如半胱胺或 我们的新型自噬刺激剂AR-13中心假设将通过以下方式进行检验： 三个具体目标：1）确定CFTR调节M Φ NOX组装的机制; 2） 确定CF特异性病原体如何差异调节M Φ ROS产生; 3）确定 新的治疗方法改变M Φ NOX/自噬轴的程度。我们将 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。这项研究之所以重要，是因为 CF巨噬细胞功能是如何调节的，这将允许新的抗生素和CFTR突变- 不可知的治疗方法感染。它也很重要，因为它将决定 特异性病原体独立地导致巨噬细胞介导的细菌杀伤的缺陷。 这项工作的预期成果将建立一个机制框架，使我们能够 靶向和纠正缺陷CF M Φ介导的细菌杀伤。最终，我们将把这个 将新知识转化为新的治疗模式，使用创新的宿主导向疗法， 对抗细菌感染。

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.