Development of an In Vitro Model of SARS-CoV-Induced Acute Lung Injury for Studying New Therapeutic Approaches

Abstract

One of the causes of death of patients infected by SARS-CoV-2 is the induced respiratory failure caused by excessive activation of the immune system, the so-called “cytokine storm”, leading to damage to lung tissue. In vitro models reproducing various stages of the disease can be used to explore the pathogenetic mechanisms and therapeutic approaches to treating the consequences of a cytokine storm. We have developed an in vitro test system for simulating damage to the pulmonary epithelium as a result of the development of a hyperinflammatory reaction based on the co-cultivation of pulmonary epithelial cells (A549 cells) and human peripheral blood mononuclear cells (PBMC) primed with lipopolysaccharide (LPS). In this model, after 24 h of co-cultivation, a sharp decrease in the rate of proliferation of A549 cells associated with the intrinsic development of oxidative stress and, ultimately, with the induction of PANoptotic death were observed. There was a significant increase in the concentration of 40 cytokines/chemokines in a conditioned medium, including TNF-α, IFN-α, IL-6, and IL-1a, which corresponded to the cytokine profile in patients with severe manifestation of COVID-19. In order to verify the model, the analysis of the anti-inflammatory effects of well-known substances (dexamethasone, LPS from Rhodobacter sphaeroides (LPS-RS), polymyxin B), as well as multipotent mesenchymal stem cells (MSC) and MSC-derived extracellular vesicles (EVs) was carried out. Dexamethasone and polymyxin B restored the proliferative activity of A549 cells and reduced the concentration of proinflammatory cytokines. MSC demonstrated an ambivalent effect through stimulated production of both pro-inflammatory cytokines and growth factors that regenerate lung tissue. LPS-RS and EVs showed no significant effect. The developed test system can be used to study molecular and cellular pathological processes and to evaluate the effectiveness of various therapeutic approaches for the correction of hyperinflammatory response in COVID-19 patients.

Research progress of extracellular vesicles and exosomes derived from mesenchymal stem cells in the treatment of oxidative stress-related diseases

“…The negative effects of ischemia-reperfusion are caused by the induction of inflammation and oxidative stress and the damage of cell energy metabolism, which leads to a series of harmful biological events from ion homeostasis failure to cell death ( 27 ). In severe acute respiratory syndrome (SARS) caused by SARS-associated coronavirus (SARS-CoV), the immune response to virus infection and cytokine storm play a key role in the severity of the disease ( 28 ). Cytokine storm will lead to the development of oxidative stress due to ROS produced by immune cells ( 29 ).…”

The protective effects of baicalin for respiratory diseases: an update and future perspectives

“…BA has been shown to inhibit SARS-CoV-2 RNA-dependent RNA polymerase activity and exhibited significant antiviral activity against SARS-CoV-2 in vitro ( Zandi et al, 2021 ). Evidence shows that cytokine storm (CS) was a pivotal stage in the advancement of COVID-19 ( Hach et al, 2022 ; Jing et al, 2022 ; Shevtsova et al, 2022 ). You et al (2022) used the method of network pharmacology to analyze the possible pathway of BA inhibiting CS.…”

hWJMSCs inhibit inflammation and apoptosis in an ARDS cell model

“…MSCs also have activity against apoptosis, microbial infections, cellular damage, and other lung injuries, as indicated by a growing number of in vivo and pre-clinical investigations. 17 In contrast, studying ARDS requires a model of cytokine storms. Inflamed lung cells are a feasible ARDS model.…”