A critical role for macrophage ferroptosis in promoting fungal invasion in lung transplant recipients

巨噬细胞铁死亡在促进肺移植受者真菌侵袭中的关键作用

• 批准号: 10186399
• 负责人: Joe L Hsu
• 金额: $ 59.3万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10186399
• 关键词: Acute; Acute Renal Failure with Renal Papillary Necrosis; Address; Adult Respiratory Distress Syndrome; Affect; Alveolar; Alveolar Macrophages; Alzheimer' s Disease; Animals; Aspergillosis; Aspergillus; Aspergillus fumigatus; Atherosclerosis; Biology; Bronchoalveolar Lavage; Cell Culture Techniques; Cell Death; Chronic Obstructive Airway Disease; Clinical; Control Animal; Cytometry; Defect; Discrimination; Effector Cell; Exposure to; Flow Cytometry; Fluorescence; Fluorescence-Activated Cell Sorting; Genes; Graft Rejection; Granulocyte-Macrophage Colony-Stimulating Factor; Hemorrhage; Histopathologic Grade; Homeostasis; Homologous Transplantation; Host Defense; Human; Immune; Immunity; In Vitro; Infection; Inflammatory; Ingestion; Innate Immune Response; Iron; Iron Chelating Agents; Iron Overload; Label; Laboratories; Life; Lipid Peroxides; Lipids; Lung Transplantation; Lung diseases; Macrophage Colony-Stimulating Factor; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Mediating; Microscopy; Molds; Morbidity - disease rate; Mus; Mycoses; Oranges; Oxidation-Reduction; Parkinson Disease; Pathogenesis; Peptide Hydrolases; Pharmaceutical Preparations; Phenotype; Procedures; Production; Protease Inhibitor; Reactive Oxygen Species; Reperfusion Injury; Reporter; Reproduction spores; Role; Savings; Statistical Methods; Techniques; Testing; Time; Tissues; Trachea; Transgenic Mice; Transplant Recipients; Transplantation; Viral Pneumonia; Western Blotting; clinically actionable; design; emerging pathogen; experimental study; improved outcome; in vivo; inhibitor/antagonist; macrophage; monocyte; mortality; nanoparticle; neutrophil; novel; novel therapeutic intervention; pathogen; post-transplant; protease E; response; response to injury; transplant model

PROJECT SUMMARY/ ABSTRACT This application focuses on the contribution of transplant hemorrhage-induced iron overload in the dysregulation of pulmonary macrophages (mɸs) and the promotion of invasive aspergillosis. Aspergillus fumigatus (Af) is a ubiquitous mold that releases airborne spores (conidia) and affects nearly 20 million people worldwide. One-in- three lung transplant recipients (LTRs) suffers from Aspergillus-related pulmonary disease. While lung transplantation can be a life-saving treatment for thousands of people, survival post-transplant is often limited by Af infection. To better understand the transplant (host)-Af (pathogen) relationship, we developed a murine orthotopic tracheal transplant (OTT) model of Af infection. We have shown that transplant rejection-mediated microhemorrhage increases tissue iron levels and determines Af invasion. However, the exact interaction between immunity, iron overload and infection are still poorly understood. Mɸs are the first line of defense against Af and are also central to restoring tissue iron homeostasis. Importantly, our preliminary results indicate that microhemorrhage-mediated iron overload: (i) profoundly impacts the ability of mfs to kill Af conidia through a defect in lysosomal acidification; (ii) the innate immune response is polarized toward a pro-inflammatory mɸs state that results in high levels of tissue damaging reactive oxygen species (ROS); and (iii) iron promotes mɸ ferroptosis. Ferroptosis is a newly recognized form of regulated cell death that results from the production of iron toxic lipid ROS. Ferroptosis was first recognized in cancer but is now known to contribute to Alzheimer’s and Parkinson’s disease, ischemia reperfusion injury, atherosclerosis, acute kidney injury and the response to acute hemorrhage. However, the role of ferroptosis in lowering the host’s defense against pathogens, if any, remains unknown. The proposed studies are designed to address these questions in terms of Af invasion. The central hypothesis is that transplant microhemorrhage-mediated iron overload induces mf ferroptosis and polarization into an unrestrained pro-inflammatory phenotype that promotes Af invasion. Specific aim 1 utilizes in vitro and in vivo experiments to investigate the concept that ferroptosis is dictated by mɸs polarization state and contributes to the inability of transplant mɸs to mitigate Af infection and studies the role of iron lowering agents and anti-ferroptotic drugs to decrease fungal invasion. Specific aim 2 uses state-of-art omics techniques to define iron induced Af proteases and tests the concept that fungal protease inhibition can mitigate ferroptosis and improve outcomes in the tracheal transplant model. Specific aim 3 studies the ability of alveolar mɸs isolated from human LTRs compared to non-lung transplants to kill Af conidia and correlates the ability of mɸs to kill conidia with mɸs-polarization state and ferroptosis, using mass cytometry. This aim will provide a direct translatability of the hypothesis that iron overload induces ferroptosis and a pro-inflammatory phenotype that promotes fungal invasion. Successful completion of these studies will allow for the discovery of a fundamental new biology and provide novel targets for the treatment of fungal infections.

项目摘要/摘要 该应用的重点是移植出血诱导的铁超负荷的贡献 肺巨噬细胞（MɸS）和促进浸润性曲霉病。曲霉菌（AF）是 无处不在的霉菌，可释放空气孢子（分生孢子），并影响全球近2000万人。 one 三名肺移植受者（LTRS）患有曲霉相关的肺部疾病。肺 移植可以是成千上万人的生命疗法，移植后的生存通常受到 AF感染。为了更好地了解移植（宿主）-af（病原体）关系，我们开发了鼠 AF感染的原位气管移植（OTT）模型。我们已经证明了移植拒绝介导的 微型射击会增加组织铁的水平并确定AF侵袭。但是，确切的相互作用 在免疫力之间，铁超负荷和感染仍然很少了解。 mɸs是针对的第一道防线 AF，也是恢复组织铁稳态的核心。重要的是，我们的初步结果表明 微含量介导的铁超载：（i）深刻影响MF通过A杀死AF分生孢子的能力 溶酶体酸化缺陷； （ii）先天免疫反应朝向促炎性MɸS 声明导致高水平的组织破坏活性氧（ROS）； （iii）铁促进mɸ 铁凋亡。铁凋亡是一种新认识的调节细胞死亡形式，是由铁的产生而导致的 有毒脂质ROS。铁凋亡最初是在癌症中识别的，但现在众所周知会导致阿尔茨海默氏症和 帕金森氏病，缺血再灌注损伤，动脉粥样硬化，急性肾脏损伤和对急性的反应 出血。但是，铁铁作用在降低宿主对病原体的防御（如果有的话）中的作用仍然存在 未知。拟议的研究旨在根据AF入侵解决这些问题。中央 假设是移植微毛发介导的铁超负荷会诱导MF螺旋病和极化 进入促进AF侵袭的不受约束的促炎表型。特定目标1利用体外和 在体内实验，以调查铁凋亡是由mɸs极化状态和 有助于移植Mɸ减轻AF感染并研究铁降低剂的作用 和抗肿瘤药物可减少真菌侵袭。特定的目标2使用最先进的Omics技术来定义 铁诱导的AF蛋白酶，并测试了真菌蛋白酶抑制可以减轻铁铁蛋白酶的概念 改善气管移植模型的预后。特定目的3研究肺泡MɸS分离的能力 与非肺移植物相比，人类LTR从人类LTR中杀死AF分生孢子并将M杀死的能力相关联 使用质量细胞仪的分生孢子，具有mɸs偏振状态和铁质吞噬作用。这个目标将提供直接 铁超负荷诱导铁铁的假设的转换性和促炎的表型 促进真菌入侵。这些研究的成功完成将允许发现基本 新的生物学并为治疗真菌感染提供了新的靶标。