Clinical lecturer University of Pennsylvania, New Bolton Center, United States
Abstract:
Background: Metabolomics studies have been performed in limited studies of horses with asthma with varrying results. In this study, mass spectrometry-based metabolomics was used to profile BALF from horses to identify metabolites related to the pathogenesis of this disease.
Hypothesis/
Objectives: We hypothesized that horses with asthma would have significant differences in metabolic profiles of BALF supernatant versus healthy. Our objective was to utilize untargeted LC/MS metabolomics to identify biomarkers in suspect asthmatic horses.
Animals: Institution-owned, and client-owned horses were enrolled. Five healthy and 5 suspect asthma horses were included.
Methods: BALF was collected from horses with suspect asthma during active disease and supernatant stored at -80 ºC. Metabolites were extracted from BALF in ice cold methanol and separated using reversed-phase C18 and HILIC chromatography coupled to a Thermo Vanquish UHPLC/Orbitrap ID-X mass spectrometer. Compound Discoverer (Thermo Scientific) was used to process raw mass spectrometry data and perform a metabolite database search. Metabolites having a Welch’s t-test p-value < 0.05 with Benjamini-Hochberg correction were considered. Post-processing involved generation of principal component analysis (PCA) plot, volcano plot, box-and-whisker plots, and heat maps.
Results: Several metabolites were identified in BALF including some that were important for group clustering and separation. Two metabolites identified that are potentially interesting in relation to airway disease are N-acetylserotonin and N-tetradecanoylsphinganine.
Conclusions and clinical importance: Identification of potential significant metabolites in horse BALF was possible utilizing LS/MS metabolomics, which may improve our understanding of equine asthma pathophysiology in follow-up studies.
