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

• 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
• 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

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 人。严重的疾病会导致急性呼吸窘迫 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 上皮细胞和内皮细胞衬里。了解这些疾病背后的机制至关重要 制定有效的预防和治疗策略。葡萄球菌肠毒素 B (SEB) 诱导 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 新冠肺炎。已经表明，与刺突相关的超抗原结构和序列 SARS-CoV-2 的蛋白质与 SEB 的蛋白质相似。这种 SEB 诱导的 ARDS 模型也导致 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 人类中有19个。当 C3H/HeJ 小鼠接受 SEB 治疗时，它们的存活率降至 0%。在我们的研究中， 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 细胞。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 表达以及巨噬细胞激活基因 Cebp。我们假设 CBD 会减弱 SEB- 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。目标1将 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 那些亚人群。目标 2 将阐明 CBD 是否影响 Cebp 表达以及其影响 miR-124-3p具有疾病的表现。目标3将确定表观遗传因素的调节 miR-124-3p 的表达。这项研究将探索 CBD 作为 ARDS 和/或的潜在治疗方法 败血症不仅由 SEB 引起，还由其他病原体（例如 SARS-CoV-2）引起。 K99R00 将提供 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。