Targeting NETosis for the Treatment of Tuberculosis

靶向 NETosis 治疗结核病

• 批准号: 10750804
• 负责人: Christina Leigh Stallings
• 金额: $ 62.96万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750804
• 关键词: Affect; Autophagocytosis; Binding; Bronchoalveolar Lavage Fluid; Cell Death Process; Cells; Cessation of life; Chemicals; Chromatin; Chromatin Structure; Complex; Cytoplasmic Granules; DNA; Data; Disease; Dissection; Genetic; Growth; Histones; Human; Immune; Immune response; In Vitro; Infection; Infection Control; Inflammation; Interferon Type I; Internet; Link; Lung; Lysosomes; Macrophage; Mediating; Mus; Mycobacterium tuberculosis; Nutrient; Pathogenesis; Pathology; Pathway interactions; Patients; Peptide Hydrolases; Phagocytosis; Phagosomes; Predisposition; Process; Production; Protein-arginine deiminase; Proteins; Reaction; Resected; Role; Severity of illness; Signal Transduction; Sputum; Structure of parenchyma of lung; Tissues; Tuberculosis; Vesicle; antimicrobial; cell type; cytokine; effective therapy; extracellular; improved; in vivo; insight; neutrophil; nonhuman primate; pathogen; response; trafficking; tuberculosis treatment; vesicular release

Abstract Neutrophils are the most abundant and predominantly-infected cell type in the sputum, bronchoalveolar lavage fluid, and caseum contents from resected lung tissue of active tuberculosis (TB) patients. Studies of TB in mice, non-human primates, and humans have identified a correlation between neutrophil abundance and increased disease severity. Although there is a growing appreciation for the association of increased neutrophil abundance with active TB disease, it was still unknown if the presence of neutrophils in the lungs of active TB patients is consequential, or if the neutrophils are bystanders reacting to an uncontrolled infection. In particular, the details on how specific neutrophil responses and effector functions impact TB disease have remained elusive. In response to Mycobacterium tuberculosis (Mtb) infection, neutrophils deploy a number of defenses including the extrusion of neutrophil extracellular traps (NETs) via a process of cell death termed NETosis. NETosis usually follows the general steps of 1) histone citrullination, 2) chromatin decondensation, and 3) release of web-like chromatin structures decorated with antimicrobial granule proteins with the potential to bind, trap, and kill pathogens. We have recently discovered that NETosis directly promotes Mtb replication, where genetic or chemical inhibition of NETosis mediates better control of Mtb infection in vitro and in vivo, thus validating NETosis as a potential target for host-directed therapies to treat TB. We have used genetic and chemical approaches to mechanistically dissect the process of NETosis during Mtb infection, which has identified a number of regulatory nodes that can be manipulated to lead to better control of Mtb pathogenesis. We find that in response to Mtb infection in neutrophils, protein arginine deiminase 4 (PAD4) citrullinates histones to decondense chromatin that gets packaged into vesicles for release as NETs in a manner that promotes Mtb replication. We discovered that type I interferon (IFN), which has been associated with NETosis in numerous contexts but without a known mechanism, promotes the formation of chromatin-containing vesicles and NET release. In addition, we discovered a new autophagy-independent role for the ATG5 protein in suppressing NETosis by blocking type I IFN-dependent induction of PAD4 activity during Mtb infection, where increased NETosis in the absence of ATG5 expression in neutrophils leads to susceptibility to Mtb. Multiple studies have linked increased levels of type I IFN signaling with TB pathology in mice and humans. Based on our data that NETosis promotes Mtb replication and pathogenesis, NETosis could contribute to the ways that type I IFN signaling impedes control of Mtb infection. In this proposal, we will dissect how NETosis is regulated during Mtb infection (Aim 1), how NETosis contributes to Mtb replication (Aim 2), and how NETosis contributes to loss of control of infection (Aim 3). By pursuing our Aims, we will identify regulatory mechanisms to be targeted by HDTs aimed at blocking NETosis during Mtb infection as well as determine what consequences result from blocking NETosis during Mtb infection.

抽象的 中性粒细胞是痰液中最丰富和主要感染的细胞类型 来自活性结核病（TB）患者切除的肺组织的液体和CASEUM含量。结核病研究 小鼠，非人类灵长类动物和人类已经确定了中性粒细胞丰度与 疾病严重程度增加。尽管对增加中性粒细胞的关联越来越多 活性结核病的丰度，尚不清楚活性结核病的肺中性粒细胞是否存在 因此，患者是结果，或者中性粒细胞是旁观者对不受控制的感染反应的。尤其， 有关特定嗜中性粒细胞反应和效应子功能如何影响结核病疾病的细节 难以捉摸。为了应对结核分枝杆菌（MTB）感染，中性粒细胞部署了许多防御 包括通过称为细胞死亡的肠道死亡过程的嗜中性粒细胞外陷阱（NET）的挤出。 Netosis通常遵循1）组蛋白柠檬酸的一般步骤，2）染色质脱凝剂和3） 释放用抗菌颗粒蛋白装饰的网络状染色质结构，有可能 绑定，陷阱并杀死病原体。我们最近发现Netosis直接促进MTB 复制，netosis的遗传或化学抑制在体外介导了MTB感染的更好控制 在体内，因此将Netosis验证为宿主定向治疗结核病的潜在靶标。我们已经使用过 遗传和化学方法在MTB感染期间机械地剖析Netosis的过程， 它已经确定了许多可以操纵的调节节点，以更好地控制MTB 发病。我们发现，为了响应中性粒细胞中的MTB感染，蛋白精氨酸脱节酶4（PAD4） 将组蛋白蛋白蛋白蛋白蛋白酶以脱发染色质，该染色质被包装到囊泡中以释放，作为网中的网 促进MTB复制的方式。我们发现了已经关联的I型I Interferon（IFN） 在众多情况下Netosis，但没有已知机制，可以促进形成 含染色质的囊泡和净释放。此外，我们发现了一个新的无自噬 ATG5蛋白在抑制Netosis中的作用，通过阻断I型IFN依赖PAD4活性的诱导 在MTB感染期间，在中性粒细胞中缺乏ATG5表达的情况下，Netosis增加导致 对MTB的敏感性。多项研究已将I型IFN信号的水平增加与结核病病理相关联 老鼠和人类。基于我们的数据，Netosis促进MTB复制和发病机理，Netosis 可能有助于I型IFN信号传导阻碍对MTB感染的控制。在这个建议中，我们 将在MTB感染期间剖析Netosis的调节方式（AIM 1），Netosis如何促进MTB复制 （AIM 2），以及Netosis如何有助于失去感染的控制（AIM 3）。通过追求我们的目标，我们将 确定旨在阻止MTB感染期间Netosis的HDT靶向的调节机制 由于确定在MTB感染期间阻止Netosis会产生什么后果。