Validation of Messenger Ribonucleic Acid Markers Differentiating Among Human Acute Respiratory Distress Syndrome Subgroups in an Ovine Model of Acute Respiratory Distress Syndrome Phenotypes

Karin Wildi; Kieran Hyslop; Jonathan Millar; Samantha Livingstone; Margaret R Passmore; Mahé Bouquet; Emily Wilson; Gianluigi LiBassi; John F Fraser; Jacky Y Suen

doi:10.3389/fmed.2022.961336

Validation of Messenger Ribonucleic Acid Markers Differentiating Among Human Acute Respiratory Distress Syndrome Subgroups in an Ovine Model of Acute Respiratory Distress Syndrome Phenotypes

Front Med (Lausanne). 2022 Jul 5:9:961336. doi: 10.3389/fmed.2022.961336. eCollection 2022.

Authors

Karin Wildi^{1

2

3}, Kieran Hyslop¹, Jonathan Millar^{1

2

4}, Samantha Livingstone^{1

2}, Margaret R Passmore^{1

2}, Mahé Bouquet^{1

2}, Emily Wilson^{1

2}, Gianluigi LiBassi^{1

2}, John F Fraser^{1

2}, Jacky Y Suen^{1

2}

Affiliations

¹ Critical Care Research Group, Brisbane, QLD, Australia.
² Faculty of Medicine, The University of Queensland, Brisbane, QLD, Australia.
³ Cardiovascular Research Group, Basel, Switzerland.
⁴ Roslin Institute, University of Edinburgh, Edinburgh, United Kingdom.

Abstract

Background: The discovery of biological subphenotypes in acute respiratory distress syndrome (ARDS) might offer a new approach to ARDS in general and possibly targeted treatment, but little is known about the underlying biology yet. To validate our recently described ovine ARDS phenotypes model, we compared a subset of messenger ribonucleic acid (mRNA) markers in leukocytes as reported before to display differential expression between human ARDS subphenotypes to the expression in lung tissue in our ovine ARDS phenotypes model (phenotype 1 (Ph1): hypoinflammatory; phenotype 2 (Ph2): hyperinflammatory).

Methods: We studied 23 anesthetized sheep on mechanical ventilation with observation times between 6 and 24 h. They were randomly allocated to the two phenotypes (n = 14 to Ph1 and n = 9 to Ph2). At study end, lung tissue was harvested and preserved in RNAlater. After tissue homogenization in TRIzol, total RNA was extracted and custom capture and reporter probes designed by NanoString Technologies were used to measure the expression of 14 genes of interest and the 6 housekeeping genes on a nCounter SPRINT profiler.

Results: Among the 14 mRNA markers, in all animals over all time points, 13 markers showed the same trend in ovine Ph2/Ph1 as previously reported in the MARS cohort: matrix metalloproteinase 8, olfactomedin 4, resistin, G protein-coupled receptor 84, lipocalin 2, ankyrin repeat domain 22, CD177 molecule, and transcobalamin 1 expression was higher in Ph2 and membrane metalloendopeptidase, adhesion G protein-coupled receptor E3, transforming growth factor beta induced, histidine ammonia-lyase, and sulfatase 2 expression was higher in Ph1. These expression patterns could be found when different sources of mRNA - such as blood leukocytes and lung tissue - were compared.

Conclusion: In human and ovine ARDS subgroups, similar activated pathways might be involved (e.g., oxidative phosphorylation, NF-κB pathway) that result in specific phenotypes.

Keywords: acute respiratory distress syndrome; biomarker; mRNA expression; phenotypes; precision medicine; predictive and prognostic enrichment; up-and downregulation.