Application across species of a one health approach to liquid sample handling for respiratory based -omics analysis

Abstract Airway inflammation is highly prevalent in horses, with the majority of non-infectious cases being defined as equine asthma. Currently, cytological analysis of airway derived samples is the principal method of assessing lower airway inflammation. Samples can be obtained by tracheal wash (TW...

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Autores principales: Anna E. Karagianni, Samantha L. Eaton, Dominic Kurian, Eugenio Cillán-Garcia, Jonathan Twynam-Perkins, Anna Raper, Thomas M. Wishart, R. Scott Pirie
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Publicado: Nature Portfolio 2021
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spelling oai:doaj.org-article:17eeacd68a0a491aa130c2281e4e09f02021-12-02T15:33:01ZApplication across species of a one health approach to liquid sample handling for respiratory based -omics analysis10.1038/s41598-021-93839-92045-2322https://doaj.org/article/17eeacd68a0a491aa130c2281e4e09f02021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-93839-9https://doaj.org/toc/2045-2322Abstract Airway inflammation is highly prevalent in horses, with the majority of non-infectious cases being defined as equine asthma. Currently, cytological analysis of airway derived samples is the principal method of assessing lower airway inflammation. Samples can be obtained by tracheal wash (TW) or by lavage of the lower respiratory tract (bronchoalveolar lavage (BAL) fluid; BALF). Although BALF cytology carries significant diagnostic advantages over TW cytology for the diagnosis of equine asthma, sample acquisition is invasive, making it prohibitive for routine and sequential screening of airway health. However, recent technological advances in sample collection and processing have made it possible to determine whether a wider range of analyses might be applied to TW samples. Considering that TW samples are relatively simple to collect, minimally invasive and readily available in the horse, it was considered appropriate to investigate whether, equine tracheal secretions represent a rich source of cells and both transcriptomic and proteomic data. Similar approaches have already been applied to a comparable sample set in humans; namely, induced sputum. Sputum represents a readily available source of airway biofluids enriched in proteins, changes in the expression of which may reveal novel mechanisms in the pathogenesis of respiratory diseases, such as asthma and chronic obstructive pulmonary disease. The aim of this study was to establish a robust protocol to isolate macrophages, protein and RNA for molecular characterization of TW samples and demonstrate the applicability of sample handling to rodent and human pediatric bronchoalveolar lavage fluid isolates. TW samples provided a good quality and yield of both RNA and protein for downstream transcriptomic/proteomic analyses. The sample handling methodologies were successfully applicable to BALF for rodent and human research. TW samples represent a rich source of airway cells, and molecular analysis to facilitate and study airway inflammation, based on both transcriptomic and proteomic analysis. This study provides a necessary methodological platform for future transcriptomic and/or proteomic studies on equine lower respiratory tract secretions and BALF samples from humans and mice.Anna E. KaragianniSamantha L. EatonDominic KurianEugenio Cillán-GarciaJonathan Twynam-PerkinsAnna RaperThomas M. WishartR. Scott PirieNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Anna E. Karagianni
Samantha L. Eaton
Dominic Kurian
Eugenio Cillán-Garcia
Jonathan Twynam-Perkins
Anna Raper
Thomas M. Wishart
R. Scott Pirie
Application across species of a one health approach to liquid sample handling for respiratory based -omics analysis
description Abstract Airway inflammation is highly prevalent in horses, with the majority of non-infectious cases being defined as equine asthma. Currently, cytological analysis of airway derived samples is the principal method of assessing lower airway inflammation. Samples can be obtained by tracheal wash (TW) or by lavage of the lower respiratory tract (bronchoalveolar lavage (BAL) fluid; BALF). Although BALF cytology carries significant diagnostic advantages over TW cytology for the diagnosis of equine asthma, sample acquisition is invasive, making it prohibitive for routine and sequential screening of airway health. However, recent technological advances in sample collection and processing have made it possible to determine whether a wider range of analyses might be applied to TW samples. Considering that TW samples are relatively simple to collect, minimally invasive and readily available in the horse, it was considered appropriate to investigate whether, equine tracheal secretions represent a rich source of cells and both transcriptomic and proteomic data. Similar approaches have already been applied to a comparable sample set in humans; namely, induced sputum. Sputum represents a readily available source of airway biofluids enriched in proteins, changes in the expression of which may reveal novel mechanisms in the pathogenesis of respiratory diseases, such as asthma and chronic obstructive pulmonary disease. The aim of this study was to establish a robust protocol to isolate macrophages, protein and RNA for molecular characterization of TW samples and demonstrate the applicability of sample handling to rodent and human pediatric bronchoalveolar lavage fluid isolates. TW samples provided a good quality and yield of both RNA and protein for downstream transcriptomic/proteomic analyses. The sample handling methodologies were successfully applicable to BALF for rodent and human research. TW samples represent a rich source of airway cells, and molecular analysis to facilitate and study airway inflammation, based on both transcriptomic and proteomic analysis. This study provides a necessary methodological platform for future transcriptomic and/or proteomic studies on equine lower respiratory tract secretions and BALF samples from humans and mice.
format article
author Anna E. Karagianni
Samantha L. Eaton
Dominic Kurian
Eugenio Cillán-Garcia
Jonathan Twynam-Perkins
Anna Raper
Thomas M. Wishart
R. Scott Pirie
author_facet Anna E. Karagianni
Samantha L. Eaton
Dominic Kurian
Eugenio Cillán-Garcia
Jonathan Twynam-Perkins
Anna Raper
Thomas M. Wishart
R. Scott Pirie
author_sort Anna E. Karagianni
title Application across species of a one health approach to liquid sample handling for respiratory based -omics analysis
title_short Application across species of a one health approach to liquid sample handling for respiratory based -omics analysis
title_full Application across species of a one health approach to liquid sample handling for respiratory based -omics analysis
title_fullStr Application across species of a one health approach to liquid sample handling for respiratory based -omics analysis
title_full_unstemmed Application across species of a one health approach to liquid sample handling for respiratory based -omics analysis
title_sort application across species of a one health approach to liquid sample handling for respiratory based -omics analysis
publisher Nature Portfolio
publishDate 2021
url https://doaj.org/article/17eeacd68a0a491aa130c2281e4e09f0
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