Role of macrophages in CBD mediated attenuation of SEB-induced ARDS

巨噬细胞在 CBD 介导的 SEB 诱导的 ARDS 减弱中的作用

基本信息

批准号：
10351483
负责人：
Kiesha Wilson
金额：
$ 10万
依托单位：
UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10351483
关键词：
2019-nCoV Acute Respiratory Distress Syndrome Address Advisory Committees Affect Alveolar Alveolar Macrophages American Amino Acid Sequence Anti-Inflammatory Agents Apoptotic Attenuated Bacteria Biological Assay Biological Markers Biological Models Biometry C3H/HeJ Mouse CCAAT-Enhancer-Binding Protein-alpha CD14 gene COVID-19 COVID-19 patient CRISPR/Cas technology CXCL9 gene Cannabidiol Cannabinoids Cannabis Cells Cellular Infiltration Characteristics Clinical Coronavirus DNA Data Disease Drops Endothelial Cells Environment Epigenetic Process Epithelial Cells Evaluation FCGR3B gene Gene Activation Gene Expression Genes Genetic Genetic Transcription Human Hypoxia Immune Immunologics Infection Inflammatory Inflammatory Response Inhalation Interleukin-1 Interleukin-1 beta Interleukin-12 Interleukin-6 Liquid substance Lung Macrophage Activation Marijuana Mediating Mentors MicroRNAs Modeling Mononuclear Multiple Organ Failure Mus Overdose Pathway Analysis Patients Persons Phagocytosis Plants Play Plethysmography Prevention strategy Preventive Property Public Health Pulmonary Inflammation Quantitative Reverse Transcriptase PCR Reporter Research Research Personnel Role SARS-CoV-2 spike protein Sepsis Severity of illness Site South Carolina Stains Staphylococcal Enterotoxin B Structure Superantigens Survival Rate TNF gene Testing Tetrahydrocannabinol Therapeutic Toxin Training Trauma Universities Up-Regulation Virus alveolar destruction attenuation base career development cell type chemokine cytokine cytokine release syndrome in silico in vivo insight interleukin-23 macrophage migration monocyte myeloid cell development neutrophil pandemic disease pathogen phytocannabinoid promoter protein structure pulmonary function receptor sedative severe COVID-19 single cell sequencing therapeutic development transcription factor

项目摘要

PROJECT SUMMARY/ABSTRACT The virus, SARS-CoV-2 has caused COVID-19 and claimed the lives of over 240,000 Americans and 1,270,000 people worldwide. Severe cases of the disease leads to Acute Respiratory Distress Syndrome (ARDS), sepsis and can be fatal due to pulmonary inflammation and destruction of the epithelial and endothelial cell lining. Understanding the mechanisms behind these diseases is vital to develop effective preventive and therapeutic strategies. Staphylococcus enterotoxin B (SEB)-induced ARDS mimics the cytokine storm, sepsis and multiple organ failure presented in patients with severe COVID-19. It has been shown that the superantigen structure and sequence associated with the spike protein of the SARS-CoV-2 is similar to that of SEB. This SEB-induced ARDS model also results in various presentations of severity of illness in mice of different genetic backgrounds, as does COVID- 19 in humans. When C3H/HeJ mice are treated with SEB, their survival rate drops to 0%. In our study, we found that Cannabidiol (CBD) administration following SEB treatment, led to 100% survival indefinitely. Initial evaluation of whole single cell sequencing data comparing lungs from naïve with SEB-induced ARDS mice illustrated that there was an increase in neutrophils, inflammatory macrophages and pro-inflammatory cytokines (IL-1β and TNF-α) as well as a loss in lung epithelial cells. To characterize the mechanism by which CBD treatment led to amelioration of the inflammatory response, microRNA expression analysis was done that showed a significant decrease in expression of miR-124-3p in SEB-treated group which is directly associated with upregulation of TNF-α and IL-1β expression as well as macrophage activation gene, Cebp. We hypothesized that CBD attenuates SEB- induced ARDS by miRNA dysregulation in lung-infiltrating cells, specifically by inducing miR-124-3p which downregulates Cebp expression resulting in reduced activation of macrophages. Aim 1 will elucidate the role of resident and monocyte-derived macrophages in disease and the effect CBD on those subpopulations. Aim 2 will elucidate whether CBD affects Cebp expression and the effects that miR-124-3p has on manifestation of disease. Aim 3 will determine the epigenetic factors regulating expression of miR-124-3p. This study will explore CBD as a potential therapeutic for ARDS and/or sepsis induced not only by SEB but other pathogens such as SARS-CoV-2. The K99R00 will provide opportunities associated with Career Development and training in –omics approaches and biostatistics. Taken together, my mentors, advisory committee, consultant, and research environment at the University of South Carolina will nurture my successful transition to an Independent Investigator.

项目概要/摘要 SARS-CoV-2 病毒引起了 COVID-19，夺去了超过 24 万美国人的生命，全球有 1,270,000 名严重病例导致急性呼吸窘迫。综合症（ARDS）、脓毒症，并且由于肺部炎症和细胞破坏而可能致命了解这些疾病背后的机制对于上皮细胞和内皮细胞衬里至关重要。制定有效的预防和治疗策略。 ARDS 模仿重症患者出现的细胞因子风暴、败血症和多器官衰竭研究表明，COVID-19 的超抗原结构和序列与刺突相关。 SARS-CoV-2 的蛋白与 SEB 的蛋白相似，这种 SEB 诱导的 ARDS 模型也会导致。不同遗传背景的小鼠的疾病严重程度有不同的表现，就像新冠病毒一样 19 在我们的研究中，当 C3H/HeJ 小鼠接受 SEB 治疗时，它们的存活率降至 0%。我们发现 SEB 治疗后给予大麻二酚 (CBD) 导致 100% 存活无限期地对全单细胞测序数据进行初步评估，比较原始肺和 SEB 诱导的 ARDS 小鼠表明中性粒细胞、炎症细胞增多巨噬细胞和促炎细胞因子（IL-1β 和 TNF-α）以及肺上皮细胞的损失描述 CBD 治疗改善炎症的机制。响应，进行了 microRNA 表达分析，结果显示表达显着下降 SEB 治疗组中 miR-124-3p 的表达，与 TNF-α 和 IL-1β 的上调直接相关我们研究了 CBD 减弱 SEB- 的表达以及巨噬细胞激活基因 Cebp。通过肺浸润细胞中的 miRNA 失调（特别是通过诱导 miR-124-3p）诱导 ARDS 下调 Cebp 表达，导致巨噬细胞活化减少。阐明常驻巨噬细胞和单核细胞衍生的巨噬细胞在疾病中的作用以及 CBD 对目标 2 将阐明 CBD 是否影响 Cebp 表达以及影响。 miR-124-3p对疾病的表现具有影响，目的3将确定调节表观遗传因素。 miR-124-3p 的表达本研究将探索 CBD 作为 ARDS 和/或的潜在治疗方法。脓毒症不仅由 SEB 引起，而且由 K99R00 等其他病原体引起。与组学方法和生物统计学的职业发展和培训相关的机会。总的来说，我的导师、咨询委员会、顾问和研究环境南卡罗来纳大学将培养我成功过渡为独立研究者。