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、支气管肺泡灌洗液中最丰富且主要感染的细胞类型 活动性结核病 (TB) 患者切除肺组织中的液体和酪蛋白内容物。结核病研究 小鼠、非人类灵长类动物和人类已经发现中性粒细胞丰度与 疾病严重程度增加。尽管人们越来越认识到中性粒细胞增多与 活动性结核病患者的肺部是否存在中性粒细胞仍不清楚 患者是后果性的，或者中性粒细胞是对不受控制的感染做出反应的旁观者。尤其， 关于特定中性粒细胞反应和效应器功能如何影响结核病的细节仍然存在 难以捉摸。为了应对结核分枝杆菌 (Mtb) 感染，中性粒细胞部署了多种防御措施 包括通过称为 NETosis 的细胞死亡过程挤出中性粒细胞胞外陷阱 (NET)。 NETosis 通常遵循以下一般步骤：1) 组蛋白瓜氨酸化，2) 染色质解缩，以及 3) 释放用抗菌颗粒蛋白装饰的网状染色质结构，有可能 结合、捕获和杀死病原体。我们最近发现 NETosis 直接促进 Mtb 复制，其中 NETosis 的遗传或化学抑制可更好地控制体外 Mtb 感染， 体内，从而验证 NETosis 作为宿主导向疗法治疗结核病的潜在靶点。我们已经用过 遗传和化学方法从机械角度剖析 Mtb 感染期间的 NETosis 过程， 它已经确定了许多可以操纵的监管节点，以更好地控制结核分枝杆菌 发病。我们发现，为了应对中性粒细胞中的 Mtb 感染，蛋白精氨酸脱亚胺酶 4 (PAD4) 瓜氨酸化组蛋白以解凝染色质，然后将其包装到囊泡中，以 NET 的形式释放 促进 Mtb 复制的方式。我们发现了 I 型干扰素 (IFN)，它与 NETosis 在许多情况下但没有已知的机制，促进了形成 含有染色质的囊泡和 NET 释放。此外，我们还发现了一种新的不依赖自噬的 ATG5 蛋白通过阻断 I 型 IFN 依赖性 PAD4 活性诱导来抑制 NETosis 的作用 在 Mtb 感染期间，中性粒细胞中缺乏 ATG5 表达的情况下 NETosis 增加导致 对结核分枝杆菌的易感性。多项研究已将 I 型干扰素信号传导水平升高与结核病病理联系起来。 老鼠和人类。根据我们的数据，NETosis 促进 Mtb 复制和发病机制，NETosis 可能有助于 I 型 IFN 信号传导阻碍 Mtb 感染的控制。在这个提案中，我们 将剖析 NETosis 在 Mtb 感染期间如何受到调节（目标 1），NETosis 如何促进 Mtb 复制 （目标 2），以及 NETosis 如何导致感染失控（目标 3）。通过追求我们的目标，我们将 确定 HDT 的目标监管机制，旨在阻止 Mtb 感染期间的 NETosis 以确定在 Mtb 感染期间阻断 NETosis 会产生什么后果。