Abstract:The rapid repurposing of antivirals is particularly pressing during pandemics. However, rapid assays for assessing candidate drugs typically involve in vitro screens and cell lines that do not recapitulate human physiology at the tissue and organ levels. Here we show that a microfluidic bronchial-airway-on-a-chip lined by highly differentiated human bronchial-airway epithelium and pulmonary endothelium can model viral infection, strain-dependent virulence, cytokine production and the recruitment of circulating… Show more
“…2A). While the presence of endothelium was shown to influence influenza virus infection in Lung Airway Chips in past studies 2,20,21 , the presence of endothelium did not significantly alter infection of intestinal epithelium by NL63 (Supp. Fig.…”
Section: Inhibition Of Nl63 Infection By Nafamostat But Not Remdesivirmentioning
confidence: 90%
“…CoV-2, in vitro 8,9,42,43 . However, the physical microenvironment of Organ chips (e.g., fluid flow, cyclic peristalsis-like mechanical deformations) has been previously shown to have potent effects on cell differentiation and function that are crucial for recapitulation of complex organ level physiology and pathophysiology with high fidelity 2,44,45 . This also seems to be the case here given that the same organoid-derived cells were cultured in the same medium in the different models, yet their phenotype differed greatly.…”
Section: Transwell Cultures Have Been Used To Study Infection By Human Coronaviruses Including Sars-mentioning
Many patients infected with coronaviruses, such as SARS-CoV-2 and NL63 that use ACE2 receptors to infect cells, exhibit gastrointestinal symptoms and viral proteins are found in the human gastrointestinal tract, yet little is known about the inflammatory and pathological effects of coronavirus infection on the human intestine. Here, we used a human intestine-on-a-chip (Intestine Chip) microfluidic culture device lined by patient organoid-derived intestinal epithelium interfaced with human vascular endothelium to study host cellular and inflammatory responses to infection with NL63 coronavirus. These organoid-derived intestinal epithelial cells dramatically increased their ACE2 protein levels when cultured under flow in the presence of peristalsis-like mechanical deformations in the Intestine Chips compared to when cultured statically as organoids or in Transwell inserts. Infection of the intestinal epithelium with NL63 on-chip led to inflammation of the endothelium as demonstrated by loss of barrier function, increased cytokine production, and recruitment of circulating peripheral blood mononuclear cells (PMBCs). Treatment of NL63 infected chips with the approved protease inhibitor drug, nafamostat, inhibited viral entry and resulted in a reduction in both viral load and cytokine secretion, whereas remdesivir, one of the few drugs approved for COVID19 patients, was not found to be effective and it also was toxic to the endothelium. This model of intestinal infection was also used to test the effects of other drugs that have been proposed for potential repurposing against SARS-CoV-2. Taken together, these data suggest that the human Intestine Chip might be useful as a human preclinical model for studying coronavirus related pathology as well as for testing of potential anti-viral or anti-inflammatory therapeutics.
“…2A). While the presence of endothelium was shown to influence influenza virus infection in Lung Airway Chips in past studies 2,20,21 , the presence of endothelium did not significantly alter infection of intestinal epithelium by NL63 (Supp. Fig.…”
Section: Inhibition Of Nl63 Infection By Nafamostat But Not Remdesivirmentioning
confidence: 90%
“…CoV-2, in vitro 8,9,42,43 . However, the physical microenvironment of Organ chips (e.g., fluid flow, cyclic peristalsis-like mechanical deformations) has been previously shown to have potent effects on cell differentiation and function that are crucial for recapitulation of complex organ level physiology and pathophysiology with high fidelity 2,44,45 . This also seems to be the case here given that the same organoid-derived cells were cultured in the same medium in the different models, yet their phenotype differed greatly.…”
Section: Transwell Cultures Have Been Used To Study Infection By Human Coronaviruses Including Sars-mentioning
Many patients infected with coronaviruses, such as SARS-CoV-2 and NL63 that use ACE2 receptors to infect cells, exhibit gastrointestinal symptoms and viral proteins are found in the human gastrointestinal tract, yet little is known about the inflammatory and pathological effects of coronavirus infection on the human intestine. Here, we used a human intestine-on-a-chip (Intestine Chip) microfluidic culture device lined by patient organoid-derived intestinal epithelium interfaced with human vascular endothelium to study host cellular and inflammatory responses to infection with NL63 coronavirus. These organoid-derived intestinal epithelial cells dramatically increased their ACE2 protein levels when cultured under flow in the presence of peristalsis-like mechanical deformations in the Intestine Chips compared to when cultured statically as organoids or in Transwell inserts. Infection of the intestinal epithelium with NL63 on-chip led to inflammation of the endothelium as demonstrated by loss of barrier function, increased cytokine production, and recruitment of circulating peripheral blood mononuclear cells (PMBCs). Treatment of NL63 infected chips with the approved protease inhibitor drug, nafamostat, inhibited viral entry and resulted in a reduction in both viral load and cytokine secretion, whereas remdesivir, one of the few drugs approved for COVID19 patients, was not found to be effective and it also was toxic to the endothelium. This model of intestinal infection was also used to test the effects of other drugs that have been proposed for potential repurposing against SARS-CoV-2. Taken together, these data suggest that the human Intestine Chip might be useful as a human preclinical model for studying coronavirus related pathology as well as for testing of potential anti-viral or anti-inflammatory therapeutics.
“…The main treatments for COVID-19 consist of palliative and supportive care. Several clinical trials have been conducted to repurpose or reposition existing drugs for COVID-19 [35,36]. From a pharmacological perspective, this is a remarkably interesting approach because it reduces the usual time for drug development, which is crucial for addressing morbidity and mortality that are caused by COVID-19 [37].…”
Section: Pathogenesis and Pathophysiology Of Covid-19mentioning
confidence: 99%
“…Interestingly, periodic mechanical motions influenced the experimental data [91]. Physiologically relevant in vitro platforms that mimic human diseases and can test the effects of drugs thereon have been highlighted by several models, such as a specific alveolar model [92], an alveolar barrier in respiratory dynamics [85], intravascular thrombosis in lung alveolus [93], small airway-on-a-chip [94], and human lung airway epithelial cells and pulmonary microvascular endothelial cells [36].…”
Section: Organ-on-a-chip and Lung-on-a-chipmentioning
Background:
Chronic respiratory diseases (CRD) are a major public health problem worldwide. In the current epidemiological context, CRD have received much interest when considering their correlation with greater susceptibility to SARS-Cov-2 and severe disease (COVID-19). Increasingly more studies have investigated pathophysiological interactions between CRD and COVID-19.
Area covered:
Animal experimentation has decisively contributed to advancing our knowledge of CRD. Considering the increase in ethical restrictions in animal experimentation, researchers must focus on new experimental alternatives. Two-dimensional (2D) cell cultures have complemented animal models and significantly contributed to advancing research in the life sciences. However, 2D cell cultures have several limitations in studies of cellular interactions. Three-dimensional (3D) cell cultures represent a new and robust platform for studying complex biological processes and are a promising alternative in regenerative and translational medicine.
Expert opinion:
Three-dimensional cell cultures are obtained by combining several types of cells in integrated and self-organized systems in a 3D structure. These 3D cell culture systems represent an efficient methodological approach in studies of pathophysiology and lung therapy. More recently, complex 3D culture systems, such as lung-on-a-chip, seek to mimic the physiology of a lung
in vivo
through a microsystem that simulates alveolar-capillary interactions and exposure to air. The present review introduces and discusses 3D lung cultures as robust platforms for studies of the pathophysiology of CRD and COVID-19 and the mechanisms that underlie interactions between CRD and COVID-19.
“…However, spontaneous transmembrane migration of airway epithelial cells might also occur 27 . A possible strategy is the concurrent seeding of epithelial and endothelial cells onto either side of the membrane to prevent such behavior 28 . Unfortunately, this method precludes the study of either compartment in isolation, constrains the type of media that can be used during epithelial differentiation, and prevents full differentiation of the epithelial cells before endothelial cells undergo senescence or suffer from suboptimal co-culture medium.…”
Human lung function is intricately linked to the mechanics of breathing; however, it remains unknown whether and how these mechanical cues shape human lung cellular biology. While respiration-related strains and fluid flows have been suggested to promote alveolar epithelial cell function, the study of such fundamental mechanisms in the conducting airway epithelium has been hindered by the lack of suitable in vitro airway models. Here, we developed a model of human bronchial airway epithelium using well-differentiated primary cell cultures on a commercial Organs-on-Chips platform that enables the application of breathing-associated airflow and cyclic strain. It furthermore features optional endothelial cell co-culture to allow for crosstalk with the vascular compartment. Using this model, we evaluated the impact of airflow and physiological levels of cyclic strain on airway epithelial cell differentiation and function. Our findings suggest that breathing-associated mechanical stimulation changes epithelial composition, reduces secretion of IL-8, and downregulates gene expression of matrix metalloproteinase 9, fibronectin, and other extracellular matrix (ECM) factors. These results indicate that breathing-associated forces are important modulators of airway epithelial cell biology and that their fine-tuned application could generate models of specific epithelial phenotypes and pathologies.
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