Strong correlation between air-liquid interface cultures and in vivo transcriptomics of nasal brush biopsy

Ghosh, Baishakhi; Park, Bong Soo; Bhowmik, Debaleena; Nishida, Kristine; Lauver, Molly; Putcha, Nirupama; Gao, Peisong; Ramanathan, Murugappan; Hansel, Nadia N.; Biswal, Shyam; Sidhaye, Venkataramana K.

doi:10.1152/ajplung.00050.2020

Cited by 27 publications

(22 citation statements)

References 11 publications

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“…Exposure to air stimulates the epithelial progenitor cells to differentiate into heterogeneous pseudostratified ciliated, goblet and basal cells, alongside other specialised cell types such as the ionocyte (Montoro et al 2018). After 28 days, this differentiated epithelium exhibits mucus production and motile cilia, and transcriptionally has been shown to strongly correlate (>96% similarity) with the expression profile of epithelial cells obtained from the original nasal brushings from the same patients (Ghosh et al 2020).…”

Section: Models Of the Airwaymentioning

confidence: 84%

Human models for COVID‐19 research

Woodall

Masonou

Case

et al. 2021

The Journal of Physiology

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Coronavirus disease 2019 (COVID-19) is an infectious disease caused by a newly emerged severe acute respiratory syndrome (SARS)-coronavirus-2 (SARS-CoV-2). COVID-19 presents a diverse clinical spectrum, ranging from an asymptomatic carrier state to patients with life-threatening multi-organ failure and death (Huang et al., 2020a). The greatest risk factor for severe disease is age, with higher morbidity and mortality rates in the elderly population, despite younger people shedding similar levels of virus (Yanez et al., 2020). The overall case fatality rate of COVID-19 is 2.3%, rising to 14.8% in patients over the age of 80, and 49% among the critically ill (Kang & Jung, 2020).Currently, therapeutics for COVID-19 are limited. To overcome this, it is important that we use physiologically relevant models to reproduce the pathology of infection and evaluate the efficacy of antiviral drugs. Models of airway infection, including the use of a human infection challenge model or well-defined, disease relevant in vitro systems can help determine the key components that perpetuate the severity of the disease. Here, we briefly review the human models that are currently being used in COVID-19 research and drug development.

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Section: Models Of the Airwaymentioning

confidence: 84%

Human models for COVID‐19 research

Woodall

Masonou

Case

et al. 2021

The Journal of Physiology

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“…However, culturing cells at the ALI is a well-established technique [53,54] in cell culture, which stimulates cilia regeneration after the native cilia have been lost [50]. In a recent study, nasal epithelial cells grown in ALI conditions were shown to well-represent their in vivo counterparts in terms of transcriptome and gene expression profile [55]. In another study, culturing nasal epithelial cells on a porous support using the ALI cell culture model stimulated the ciliary differentiation of nasal epithelial cells.…”

Section: Discussionmentioning

confidence: 99%

Preliminary Study on the Development of In Vitro Human Respiratory Epithelium Using Collagen Type I Scaffold as a Potential Model for Future Tracheal Tissue Engineering

et al. 2021

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Pathological conditions of the tracheal epithelium, such as postoperative injuries and chronic conditions, often compromise the functionality of the respiratory epithelium. Although replacement of the respiratory epithelium using various types of tracheal transplantation has been attempted, there is no predictable and dependable replacement method that holds for safe and practicable long-term use. Therefore, we used a tissue engineering approach for ex vivo regeneration of the respiratory epithelium (RE) construct. Collagen type I was isolated from sheep tendon and it was fabricated in a three-dimensional (3D) scaffold format. Isolated human respiratory epithelial cells (RECs) and fibroblasts from nasal turbinate were co-cultured on the 3D scaffold for 48 h, and epithelium maturation was allowed for another 14 days in an air–liquid interface culture system. The scanning electron microscope results revealed a fabricated porous-structure 3D collagen scaffold. The scaffold was found to be biocompatible with RECs and fibroblasts and allows cells attachment, proliferation, and migration. Immunohistochemical analysis showed that the seeded RECs and fibroblasts were positive for expression of cytokeratin 14 and collagen type I markers, respectively, indicating that the scaffold supports the native phenotype of seeded cells over a period of 14 days. Although a longer maturation period is needed for ciliogenesis to occur in RECs, the findings suggest that the tissue-engineered RE construct is a potential candidate for direct use in tracheal epithelium replacement or tracheal tube reengineering.

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“…The cryopreserved cells were expanded and maintained at an air–liquid interface (ALI) as described previously ( Nishida et al, 2017 ; Ghosh et al, 2020 ). The cryopreserved cells were amplified on rat tail collagen I (Corning, NY, USA)-coated flasks with PneumaCult TM -Ex Plus medium (StemCell Technologies Inc., Vancouver, Canada).…”

Section: Methodsmentioning

confidence: 99%

Epithelial plasticity in COPD results in cellular unjamming due to an increase in polymerized actin

Ghosh

Nishida

Chandrala

et al. 2022

Journal of Cell Science

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The airway epithelium is subjected to insults such as cigarette smoke (CS), a primary cause of Chronic Obstructive Pulmonary Disease (COPD) and serves as an excellent model to study cell plasticity. Both CS-exposed and COPD-patient derived epithelia (CHBE) display quantitative evidence of cellular plasticity, with loss of specialized apical features and a transcriptional profile suggestive of partial epithelial to mesenchymal transition, albeit with distinct cell motion indicative of cellular unjamming. These injured/diseased cells have an increased fraction of polymerized actin, due to loss of the actin-severing protein, cofilin-1. Decreasing polymerized actin restores the jammed state in both CHBE and CS exposed epithelia, indicating that the fraction of polymerized actin is critical in unjamming the epithelia. Kinetic energy spectral analysis suggests that loss of cofilin-1 results in unjamming, similar to that seen with both CS exposure and in CHBE cells. Our data suggest that in response to chronic injury, although epithelial cells display evidence of pEMT, their movement is more consistent with cellular unjamming. Inhibitors of actin polymerization rectify the unjamming features of the monolayer.

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Strong correlation between air-liquid interface cultures and in vivo transcriptomics of nasal brush biopsy

Cited by 27 publications

References 11 publications

Human models for COVID‐19 research

Human models for COVID‐19 research

Preliminary Study on the Development of In Vitro Human Respiratory Epithelium Using Collagen Type I Scaffold as a Potential Model for Future Tracheal Tissue Engineering

Epithelial plasticity in COPD results in cellular unjamming due to an increase in polymerized actin

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