Role of macrophages and miRNA in regulating lung macrophage polarization and lung pathogenesis during respiratory virus-induced acute lung injury in normal and diabetic Syrian hamsters.

正常和糖尿病叙利亚仓鼠呼吸道病毒引起的急性肺损伤期间巨噬细胞和 miRNA 在调节肺巨噬细胞极化和肺部发病机制中的作用。

• 批准号: 10701207
• 负责人: Jay R Radke
• 金额: --
• 依托单位: BOISE VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10701207
• 关键词: 2019-nCoV; Acute Lung Injury; Acute Respiratory Distress Syndrome; Adenovirus Protein; Adenoviruses; Affect; Alveolar Macrophages; Animal Model; Anti-Inflammatory Agents; Blood Glucose; Bronchopneumonia; COVID-19; COVID-19 severity; Cells; Cessation of life; Clinical; Communicable Diseases; Comparative Study; Data; Development; Diabetes Mellitus; Diet; Disease; Disease Outbreaks; Etiology; Fatty acid glycerol esters; Functional disorder; Future; Gene Expression; General Population; Genes; Glycolysis; Goals; Hamsters; Hospital Mortality; Hospitalization; Human; Immune response; Impairment; In Vitro; Incidence; Individual; Infection; Infection prevention; Inflammation; Inflammatory; Inflammatory Response; Influenza; Influenza A Virus, H1N1 Subtype; Innate Immune Response; Lung; Lung infections; Macrophage; Mediating; Mesocricetus auratus; Metabolic; Metabolic Diseases; Metabolic Pathway; MicroRNAs; Modeling; Molecular; Morbidity - disease rate; Non-Insulin-Dependent Diabetes Mellitus; Outcome; Pathogenesis; Pathogenicity; Patients; Pattern; Phenotype; Play; Pneumonia; Production; Proteomics; Pulmonary Inflammation; Reporting; Repression; Resistance; Resolution; Risk; Rodent; Role; SARS coronavirus; SARS-CoV-2 infection; Seasons; Signal Repression; Signal Transduction Pathway; Structure; System; Testing; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Veterans; Viral; Viral Pathogenesis; Viral Respiratory Tract Infection; Virus; Virus Diseases; candidate identification; cell growth regulation; chemokine; comparative; coronavirus disease; cytokine; design; diabetic; diabetic patient; effective therapy; experimental study; high risk; immune cell infiltrate; immunoregulation; in vivo; influenza virus strain; innovation; lung injury; metabolomics; mortality; multiple omics; new therapeutic target; non-diabetic; novel; novel therapeutic intervention; pandemic disease; permissiveness; prevent; prototype; pulmonary function; respiratory; respiratory virus; response; single-cell RNA sequencing; sugar; transcriptomics; virology; western diet

Over 25% of US Veterans have diabetes, and those Veterans are at an increased risk of hospitalization and increased morbidity/mortality following severe respiratory viral infections, such as, influenza (H1N1), SARS- CoV-2 (COVID) and adenovirus (Ad). Infection with these respiratory viruses causes acute lung injury (ALI) that can result in acute respiratory distress syndrome (ARDS), with a mortality rate of ~40%. There are few therapeutic options for ALI/ARDS. Virus induced ALI/ARDS is driven primarily by uncontrolled inflammatory responses. Alveolar macrophages both induce and resolve ALI/ARDS, based on their polarization/inflammatory state. The plasticity of macrophages to vary between pro-inflammatory (M1, pro-ALI/ARDS) and anti- inflammatory (M2, anti-ALI/ARDS) phenotypes is driven by their metabolic states. Diabetes is a metabolic disorder in which levels of blood glucose are high and glycolysis is the preferred cellular metabolic pathway. Macrophages from diabetic patients have a high rate of glycolysis and an increased M1 phenotype. In addition, macrophages from diabetic patients have a lower rate of plasticity to change from M1 to M2 because of this shift to glycolysis. One possibility is that this glycolytic shift contributes to severe outcomes from respiratory viral infections in diabetic patients. The Syrian hamster is naturally permissive for influenza, SARS-CoV-2 and Ad (in contrast to other rodents that require viral adaptation). In addition, the Syrian hamster can naturally become diabetic with a high fat/high sugar diet. Ad14p1 is an emergent strain of Ad14 that has caused outbreaks of severe respiratory illness and ALI/ARDS throughout the world. Hamster infection with Ad14p1 results in a patchy bronchopneumonia, as seen in other severe human viral respiratory infections. In contrast, the prototype strain of Ad14 induces little lung inflammation. Other studies have shown that cells dying from Ad14 infection induce an M2-like human macrophage response, while cells dying from Ad14p1 infection fail to change M1 alveolar macrophages to an M2 phenotype. This dying infected cell activity is regulated by the expression of the Ad gene, E1B 20K. Cells infected by Ad14 produced sufficient E1B 20K to repolarize M1 macrophages to M2, while Ad14p1 infection does not produce sufficient E1B 20K, and the infected cells fail to alter M1 macrophage polarization. Therefore, the hamster model of Ad14p1 ALI/ARDS provides an appropriate system to study how diabetes affects macrophage polarization and pathogenesis during severe viral respiratory infections. The long-term goal of this project is to understand how emergent viruses regulate macrophage polarization to develop novel therapeutic strategies to drive macrophage polarization to an ALI/ARDS resolving phenotype in both diabetic and non-diabetic Veterans. To achieve this goal, a multi-omics approach will be used to identify and phenotype macrophages in normal and diabetic hamsters infected with Ad14p1. Transcriptomics using single-cell RNA sequencing will use gene expression profiles at the resolution of individual cells to identify and phenotype macrophages and their polarization states. Infiltrating immune cells and other lung resident cells will also be identified. Proteomics will be used to identify cytokines and chemokines that drive Ad14p1 pathogenesis. Metabolomics will be used to understand the unique metabolic changes in the lungs during Ad14p1 infection in diabetes and how those changes affect macrophage polarization. Comparative virology studies with infection of normal and diabetic hamsters with a pandemic strain of H1N1 influenza will be used to determine whether similar mechanisms of pathogenesis are involved in ALI/ARDS pathogenesis induced by other severe respiratory viruses. Finally, we will test the role of miRNA expression during prototype Ad14 and Ad14p1 infection in regulating macrophage polarization and pathogenesis, with the goals of defining mechanisms of immunomodulation and identifying candidate miRNAs that might be used as therapeutic agents against viral ALI/ARDS.

超过25％的美国退伍军人患有糖尿病，那些退伍军人的住院风险增加 并增加了严重呼吸道病毒感染后的发病率/死亡率，例如流感（H1N1），SARS- COV-2（COVID）和腺病毒（AD）。这些呼吸道病毒感染导致急性肺损伤（ALI） 可能导致急性呼吸窘迫综合征（ARDS），死亡率约为40％。很少 ALI/ARDS的治疗选择。病毒诱导的ALI/ARDS主要由不受控制的炎症驱动 回答。肺泡巨噬细胞基于其极化/炎症诱导和解决ALD 状态。巨噬细胞的可塑性在促炎（M1，Pro-Ali/ards）和抗 - 之间有所不同 炎症（M2，抗ARD/ARDS）表型由其代谢状态驱动。糖尿病是一种代谢 血糖水平高且糖酵解的疾病是首选的细胞代谢途径。 来自糖尿病患者的巨噬细胞的糖酵解率很高，M1表型增加。此外， 糖尿病患者的巨噬细胞的可塑性速度较低，因为这种转变是从M1变为M2 糖酵解。一种可能性是这种糖酵解转移导致呼吸道病毒的严重结果 糖尿病患者的感染。 叙利亚仓鼠自然允许流感，SARS-COV-2和AD（与其他 需要病毒适应的啮齿动物）。此外，叙利亚仓鼠自然可以糖尿病 脂肪/高糖饮食。 AD14P1是一种新兴的AD14菌株，导致严重呼吸道疾病的爆发 和全世界的Ali/Ards。 AD14P1感染的仓鼠感染会导致斑驳的支气管瘤，AS 在其他严重的人类病毒呼吸道感染中可见。相反，AD14的原型应变很少诱导 肺部炎症。其他研究表明，从AD14感染中死亡的细胞会诱导M2样人 巨噬细胞反应，而从AD14P1感染死亡的细胞无法将M1肺泡巨噬细胞改变为 M2表型。这种垂死的感染细胞活性受AD基因E1B 20K的表达调节。细胞 被AD14感染可产生足够的E1b 20K，使M1巨噬细胞重磨到M2，而AD14P1感染确实 无法产生足够的E1B 20K，并且被感染的细胞无法改变M1巨噬细胞极化。因此， AD14P1 ALI/ARDS的仓鼠模型提供了一个适当的系统来研究糖尿病如何影响巨噬细胞 严重病毒呼吸道感染期间的极化和发病机理。该项目的长期目标是 了解新兴病毒如何调节巨噬细胞极化以开发新的治疗策略 将巨噬细胞极化驱动到糖尿病和非糖尿病退伍军人的ALI/ARDS分辨表型。 为了实现这一目标，将使用多词的方法来识别和表型巨噬细胞 受AD14P1感染的正常和糖尿病仓鼠。使用单细胞RNA测序的转录组学将使用 基因表达在单个细胞的分辨率下以识别和表型巨噬细胞及其 极化状态。还将鉴定浸润的免疫细胞和其他肺部驻留细胞。蛋白质组学会 用于鉴定驱动AD14P1发病机理的细胞因子和趋化因子。代谢组学将用于 了解糖尿病中AD14P1感染期间肺部独特的代谢变化以及它们如何 变化影响巨噬细胞极化。正常和糖尿病感染的比较病毒学研究 患有H1N1流感流行病的仓鼠将用于确定是否具有相似的机制 发病机理参与其他严重呼吸道病毒诱导的ALI/ARDS发病机理。最后，我们 将测试miRNA表达在原型AD14和AD14P1感染中的作用在调节巨噬细胞中的作用 极化和发病机理，其目标是定义免疫调节机制并识别 候选miRNA，可以用作针对病毒ali/ards的治疗剂。