Identification of urban particulate matter-induced disruption of human respiratory mucosa integrity using whole transcriptome analysis and organ-on-a chip

Abstract

BackgroundExposure to air particulate matter (PM) is associated with various diseases in the human respiratory system. To date, most in vitro studies showing cellular responses to PM have been performed in cell culture using a single cell type. There are few studies considering how multicellular networks communicate in a tissue microenvironment when responding to the presence of PM. Here, an in vitro three-dimensional (3D) respiratory mucosa-on-a-chip, composed of human nasal epithelial cells, fibroblasts, and endothelial cells, is used to recapitulate and better understand the effects of urban particulate matter (UPM) on human respiratory mucosa.ResultsWe hypothesized that the first cells to contact with UPM, the nasal epithelial cells, would respond similar to the tissue microenvironment, and the 3D respiratory mucosa model would be a suitable platform to capture these events. First, whole transcriptome analysis revealed that UPM induced gene expression alterations in inflammatory and adhesion-related genes in human nasal epithelial cells. Next, we developed an in vitro 3D respiratory mucosa model composed of human nasal epithelial cells, fibroblasts, and endothelial cells and demonstrated that the model is structurally and functionally compatible with the respiratory mucosa. Finally, we used our model to expose human nasal epithelial cells to UPM, which led to a disruption in the integrity of the respiratory mucosa by decreasing the expression of zonula occludens-1 in both the epithelium and endothelium, while also reducing vascular endothelial cadherin expression in the endothelium.ConclusionsWe demonstrate the potential of the 3D respiratory mucosa model as a valuable tool for the simultaneous evaluation of multicellular responses caused by external stimuli in the human respiratory mucosa. We believe that the evaluation strategy proposed in the study will move us toward a better understanding of the detailed molecular mechanisms associated with pathological changes in the human respiratory system.

Exosomal miRNA-19a and miRNA-614 Induced by Air Pollutants Promote Proinflammatory M1 Macrophage Polarization via Regulation of RORα Expression in Human Respiratory Mucosal Microenvironment

“…The primary human nasal epithelial cells (pHNEs) were harvested by scraping from the human inferior turbinate of patients during augmentation rhinoplasty and cultured in airway epithelial cell growth medium (Pro-moCell, Heidelberg, Germany) after being treated with dispase for 24 h at 4˚C (28). We collected a total of nine pairs of normal and CRS patient samples from Korea University Hospital.…”

“…Low-quality raw reads were eliminated, and data were aligned to Homo sapiens (hg19) using HISAT v2.0.5. The sorted readings were configured to estimate their abundance as fragments per kilobase of exon per million fragments mapped values of the transcript and gene expressed in each sample using StringTie v1.3.3b (28).…”

Human mini-brains for reconstituting central nervous system disorders

“…7d). 126 However, further effective models adding the brain component and reconstituting the OB–brain axis are needed to clarify the underlying mechanisms of OB-driven pathogenesis leading to CNS disorders in the future as well.…”

Models for barrier understanding in health and disease in lab-on-a-chips

“…Exposure to air pollution and particulate matter (PM) is associated with various respiratory diseases in humans. However, there are few studies exploring how multicellular networks communicate within a tissue microenvironment after PM exposure 34 . Byun et al developed a three-dimensional (3D) respiratory mucosa-on-a-chip model in vitro, composed of human nasal epithelial cells, fibroblasts, and endothelial cells, to investigate the effects of urban particulate matter (UPM) on the human respiratory mucosa ( Figure 5 ) 34 .…”