Immunometabolic effect of nitric oxide on human macrophages challenged with the SARS-CoV2-induced cytokine storm

Acute bilateral pneumonia in individuals infected with COVID-19 is the result of a storm of pro-inflammatory cytokines and proteases intensively released by alveolar macrophages, recruiting neutrophils and promoting 'netosis'. Both processes are responsible for the subsequent damage (severe respiratory distress, neutropenia, leukopenia, humoral elevation of inflammation mediators, etc.), which spreads to other organs (thromboembolism, liver, kidney and heart damage). This proposal aims at the administration of inhaled nitric oxide (NO) and NO donors at the systemic level in order to prevent the activation of the innate immune system in the lung, as well as to avoid neutropenia and thromboembolism phenomena, preserving the adaptative immune system function for the benefit of the patient. For this purpose, the immunometabolic properties of human-derived macrophages treated with pro-inflammatory cytokines and an NO donor were evaluated. mRNA profiles of primary cultures of human monocyte-derived macrophages treated with CK (IL1β, IL-6, IL8, TNFα, IFNƴ and GM-CSF) 20ng/mL for 24h, DETANONOATE 500µM (D500µM) 24h or CK (1h)+DETANONOATE 500µM 24h (D500µM) in RPMI+2%FBS or vehicle (controls RPMI+2%FBS) **NOTE: the GSM7527763, GSM7527764 sample treatment information has been updated on Oct 13, 2024****

感染新型冠状病毒（COVID-19）的个体所出现的急性双侧肺炎，由肺泡巨噬细胞大量释放促炎细胞因子与蛋白酶引发的细胞因子风暴导致。该风暴会招募中性粒细胞并促进中性粒细胞胞外陷阱形成（netosis），上述两种过程均会介导后续组织损伤，包括重度呼吸窘迫、中性粒细胞减少症、白细胞减少症、炎症介质体液水平升高等，并可波及其他器官，引发血栓栓塞、肝、肾及心脏损伤。本研究拟通过全身性给药吸入型一氧化氮（nitric oxide, NO）及NO供体，以期阻断肺部先天免疫系统的激活，同时避免中性粒细胞减少与血栓栓塞事件，保留适应性免疫系统功能，最终使患者获益。为此，本研究评估了经促炎细胞因子与NO供体处理的人源巨噬细胞的免疫代谢特性。具体实验分组如下：将人单核细胞来源的巨噬细胞原代培养物分别以20ng/mL的细胞因子混合物（IL1β、IL-6、IL8、TNFα、IFNγ及GM-CSF，简称CK）处理24小时；以500μM的DETANONOATE（简称D500μM）处理24小时；或先以CK预处理1小时，再以500μM DETANONOATE处理24小时；对照组则采用RPMI培养基+2%胎牛血清（Fetal Bovine Serum, FBS）或溶剂处理。**注：GSM7527763、GSM7527764样本的处理信息已于2024年10月13日更新**