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.

项目总结/摘要 这种病毒，SARS-CoV-2，已经导致COVID-19，并夺去了超过24万美国人的生命， 全球127万人。严重病例会导致急性呼吸窘迫 急性呼吸窘迫综合征（ARDS），败血症，并可能是致命的，由于肺部炎症和破坏的 上皮和内皮细胞衬里。了解这些疾病背后的机制对于 制定有效的预防和治疗战略。葡萄球菌肠毒素B（SE B）诱导 急性呼吸窘迫综合征（ARDS）与严重急性呼吸窘迫综合征（ARDS）患者的细胞因子风暴、脓毒症和多器官功能衰竭相似。 2019冠状病毒病。已经表明，与刺突相关的超抗原结构和序列 SARS-CoV-2蛋白与SEB蛋白相似。这种SEB诱导的ARDS模型还导致 不同遗传背景的小鼠中疾病严重程度的各种表现，COVID-1也是如此。 19在人类当用SEB治疗C3 H/HeJ小鼠时，它们的存活率下降到0%。在我们的研究中， 我们发现，SEB治疗后给予大麻二酚（CBD）， 无限期比较来自初治肺与来自非初治肺的全单细胞测序数据的初步评价 SEB诱导的ARDS小鼠表明，中性粒细胞，炎性细胞， 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 细胞为了表征CBD治疗导致炎症改善的机制， 响应，进行microRNA表达分析，显示表达显著降低， SEB处理组中miR-124- 3 p的表达与TNF-α和IL-1β的上调直接相关 表达以及巨噬细胞活化基因Cebp.我们假设CBD减弱SEB- 通过肺浸润细胞中的miRNA失调诱导ARDS，特别是通过诱导miR-124- 3 p 其下调Cebp表达，导致巨噬细胞活化减少。目标1将 阐明常驻和单核细胞衍生的巨噬细胞在疾病中的作用以及CBD对 这些亚群。目的2将阐明CBD是否影响Cebp的表达， miR-124- 3 p没有疾病表现。目的3将确定表观遗传因素调节 miR-124- 3 p的表达。本研究将探索CBD作为ARDS和/或 脓毒症不仅由SEB引起，还由其他病原体如SARS-CoV-2引起。K99 R 00将提供 与职业发展和组学方法和生物统计学培训相关的机会。 总的来说，我的导师，咨询委员会，顾问和研究环境在 南卡罗来纳州大学将培养我成功地过渡到一个独立的调查员。