Search

Introduction: Asthma is a heterogeneous disease with poorly defined phenotypes. Patients with severe asthma often receive multiple treatments including oral corticosteroids (OCS). Treatment may modify the observed metabotype, rendering it challenging to investigate underlying disease mechanisms. Here, we aimed to identify dysregulated metabolic processes in relation to asthma severity and medication., Methods: Baseline urine was collected prospectively from healthy participants (n=100), patients with mild-to-moderate asthma (n=87) and patients with severe asthma (n=418) in the cross-sectional U-BIOPRED cohort; 12-18-month longitudinal samples were collected from patients with severe asthma (n=305). Metabolomics data were acquired using high-resolution mass spectrometry and analysed using univariate and multivariate methods., Results: A total of 90 metabolites were identified, with 40 significantly altered (p<0.05, false discovery rate <0.05) in severe asthma and 23 by OCS use. Multivariate modelling showed that observed metabotypes in healthy participants and patients with mild-to-moderate asthma differed significantly from those in patients with severe asthma (p=2.6×10 -20 ), OCS-treated asthmatic patients differed significantly from non-treated patients (p=9.5×10 -4 ), and longitudinal metabotypes demonstrated temporal stability. Carnitine levels evidenced the strongest OCS-independent decrease in severe asthma. Reduced carnitine levels were associated with mitochondrial dysfunction via decreases in pathway enrichment scores of fatty acid metabolism and reduced expression of the carnitine transporter SLC22A5 in sputum and bronchial brushings., Conclusions: This is the first large-scale study to delineate disease- and OCS-associated metabolic differences in asthma. The widespread associations with different therapies upon the observed metabotypes demonstrate the need to evaluate potential modulating effects on a treatment- and metabolite-specific basis. Altered carnitine metabolism is a potentially actionable therapeutic target that is independent of OCS treatment, highlighting the role of mitochondrial dysfunction in severe asthma., Competing Interests: Conflict of interest: S.N. Reinke reports grants from Canadian Institutes of Health Research, during the conduct of the study. Conflict of interest: S. Naz has nothing to disclose. Conflict of interest: R. Chaleckis has nothing to disclose. Conflict of interest: H. Gallart-Ayala has nothing to disclose. Conflict of interest: J. Kolmert reports personal fees for consultancy from Gesynta Pharma AB, outside the submitted work. Conflict of interest: N.Z. Kermani has nothing to disclose. Conflict of interest: A. Tiotiu has nothing to disclose. Conflict of interest: D.I. Broadhurst has nothing to disclose. Conflict of interest: A. Lundqvist has nothing to disclose. Conflict of interest: H. Olsson is an employee and shareholder of AstraZeneca. Conflict of interest: M. Ström has nothing to disclose. Conflict of interest: Å.M. Wheelock has nothing to disclose. Conflict of interest: C. Gómez has nothing to disclose. Conflict of interest: M. Ericsson has nothing to disclose. Conflict of interest: A.R. Sousa has nothing to disclose. Conflict of interest: J.H. Riley works for and own shares in GlaxoSmithKline. Conflict of interest: S. Bates is an employee of Johnson & Johnson and has previously worked for and holds stock in GlaxoSmithKline. Conflict of interest: J. Scholfield reports grants from Innovative Medicines Initiative, during the conduct of the study; and is director and employee of TopMD Precision Medicine Ltd. Conflict of interest: M. Loza is an employee of and owns stock in Johnson & Johnson. Conflict of interest: F. Baribaud is a shareholder of Johnson & Johnson and a current employee of Bristol Myers Squibb. Conflict of interest: P.S. Bakke reports personal fees for advisory board work and lectures from AstraZeneca, and personal fees for lectures from Novartis and Boehringer Ingelheim, outside the submitted work. Conflict of interest: M. Caruso has nothing to disclose. Conflict of interest: P. Chanez reports grants and personal fees from AstraZeneca, ALK, Boehringer Ingelheim, Chiesi, Sanofi-Aventis, Novartis and GlaxoSmithKline, outside the submitted work. Conflict of interest: S.J. Fowler reports personal fees from AstraZeneca, Novartis, TEVA and Chiesi, outside the submitted work. Conflict of interest: T. Geiser has nothing to disclose. Conflict of interest: P. Howarth has nothing to disclose. Conflict of interest: I. Horvath has nothing to disclose. Conflict of interest: N. Krug has nothing to disclose. Conflict of interest: P. Montuschi has nothing to disclose. Conflict of interest: A. Behndig has nothing to disclose. Conflict of interest: F. Singer reports personal fees from Vertex Pharmaceuticals (CH) and Novartis, outside the submitted work. Conflict of interest: J. Musial has nothing to disclose. Conflict of interest: D.E. Shaw has nothing to disclose. Conflict of interest: B. Dahlén reports personal fees for advisory board work and lectures from AstraZeneca, TEVA and Sanofi, and grants from Novartis and GlaxoSmithKline, outside the submitted work. Conflict of interest: S. Hu has nothing to disclose. Conflict of interest: J. Lasky-Su has nothing to disclose. Conflict of interest: P.J. Sterk reports a public private grant from the Innovative Medicines Initiative (IMI) covered by the EU and EFPIA, during the conduct of the study. Conflict of interest: K.F. Chung has received honoraria for participating in advisory board meetings of GlaxoSmithKline, AstraZeneca, Roche, Novartis, Merck, Nocion and Shionogi regarding treatments for asthma, COPD and chronic cough and has also been remunerated for speaking engagements. Conflict of interest: R. Djukanovic reports receiving fees for lectures at symposia organised by Novartis, AstraZeneca and TEVA, consultation for TEVA and Novartis as member of advisory boards, and participation in a scientific discussion about asthma organised by GlaxoSmithKline; and is a co-founder and current consultant, and has shares in Synairgen, a University of Southampton spin out company. Conflict of interest: S-E. Dahlén reports personal fees for consultancy from AstraZeneca, Cayman Chemical, GlaxoSmithKline, Novartis, Merck, Regeneron, Sanofi and TEVA, outside the submitted work. Conflict of interest: I.M. Adcock has nothing to disclose. Conflict of interest: C.E. Wheelock has nothing to disclose., (Copyright ©The authors 2022.)

Rationale: Asthma phenotyping requires novel biomarker discovery., Objectives: To identify plasma biomarkers associated with asthma phenotypes by application of a new proteomic panel to samples from two well-characterised cohorts of severe (SA) and mild-to-moderate (MMA) asthmatics, COPD subjects and healthy controls (HCs)., Methods: An antibody-based array targeting 177 proteins predominantly involved in pathways relevant to inflammation, lipid metabolism, signal transduction and extracellular matrix was applied to plasma from 525 asthmatics and HCs in the U-BIOPRED cohort, and 142 subjects with asthma and COPD from the validation cohort BIOAIR. Effects of oral corticosteroids (OCS) were determined by a 2-week, placebo-controlled OCS trial in BIOAIR, and confirmed by relation to objective OCS measures in U-BIOPRED., Results: In U-BIOPRED, 110 proteins were significantly different, mostly elevated, in SA compared to MMA and HCs. 10 proteins were elevated in SA versus MMA in both U-BIOPRED and BIOAIR (alpha-1-antichymotrypsin, apolipoprotein-E, complement component 9, complement factor I, macrophage inflammatory protein-3, interleukin-6, sphingomyelin phosphodiesterase 3, TNF receptor superfamily member 11a, transforming growth factor-β and glutathione S-transferase). OCS treatment decreased most proteins, yet differences between SA and MMA remained following correction for OCS use. Consensus clustering of U-BIOPRED protein data yielded six clusters associated with asthma control, quality of life, blood neutrophils, high-sensitivity C-reactive protein and body mass index, but not Type-2 inflammatory biomarkers. The mast cell specific enzyme carboxypeptidase A3 was one major contributor to cluster differentiation., Conclusions: The plasma proteomic panel revealed previously unexplored yet potentially useful Type-2-independent biomarkers and validated several proteins with established involvement in the pathophysiology of SA., Competing Interests: Conflict of interest: M. Sparreman Mikus has nothing to disclose. Conflict of interest: J. Kolmert reports personal fees from Gesynta Pharma AB. Conflict of interest: L.I. Andersson has nothing to disclose. Conflict of interest: J. Östling has nothing to disclose. Conflict of interest: R.G. Knowles has nothing to disclose. Conflict of interest: C. Gómez has nothing to disclose. Conflict of interest: M. Ericsson has nothing to disclose. Conflict of interest: J-O. Thörngren has nothing to disclose. Conflict of interest: P. Emami Khoonsari has nothing to disclose. Conflict of interest: B. Dahlén reports personal fees from AstraZeneca, Teva, Sanofi and grants from Novartis and GlaxoSmithKline outside the submitted work. Conflict of interest: M. Kupczyk has nothing to disclose. Conflict of interest: B. De Meulder report grants from the Innovative Medicines Initiative during the conduct of the study. Conflict of interest: C. Auffray report grants from the Innovative Medicines Initiative during the conduct of the study. Conflict of interest: P.S. Bakke reports personal fees from GlaxoSmithKline, AstraZeneca, Boehringer Ingelheim and Chiesi outside the submitted work. Conflict of interest: B. Beghe reports personal fees from AstraZeneca, Boehringer Ingelheim, Menarini and GlaxoSmithKline outside the submitted work. Conflict of interest: E.H. Bel reports grants and personal fees from GlaxoSmithKline, AstraZeneca, Novartis, TEVA, Sanofi/Regeneron, Chiesi, and Sterna outside the submitted work. Conflict of interest: M. Caruso has nothing to disclose. Conflict of interest: P. Chanez has nothing to disclose. Conflict of interest: B. Chawes has nothing to disclose. Conflict of interest: S.J. Fowler has nothing to disclose. Conflict of interest: M. Gaga reports grants and personal fees from Novartis, Menarini, Merck Sharp & Dohme, BMS, Galapagos, and AstraZeneca outside the submitted work. Conflict of interest: T. Geiser has nothing to disclose. Conflict of interest: M. Gjomarkaj has nothing to disclose. Conflict of interest: I. Horváth reports grants from EFPIA during the conduct of the study, and personal fees from AstraZeneca, GlaxoSmithKline, Novartis, Boehringer-Ingelheim, Sandoz, Teva and Chiesi outside the submitted work. Conflict of interest: P.H. Howarth has nothing to disclose. Conflict of interest: S.L. Johnston reports personal fees from Virtus Respiratory Research, Myelo Therapeutics GmbH, Concert Pharmaceuticals, Bayer, Synairgen, Novartis, Boehringer Ingelheim, Chiesi, Gerson Lehrman Group, resTORbio, Bioforce, Materia Medical Holdings, PrepBio Pharma, Pulmotect, Virion Health, Lallemand Pharma and AstraZeneca outside the submitted work. In addition, Sebastian L. Johnston also has three patents (Anti-virus therapy for respiratory diseases, UK patent application No. GB 0405634.7, Interferon-Beta for Anti-Virus Therapy for Respiratory Diseases, International Patent Application No. PCT/ GB05/50031 and Interferon Lambda therapy for the treatment of respiratory disease, UK patent application No.6779645.9). Conflict of interest: G. Joos reports grants and personal fees from AstraZeneca, Bayer, Chiesi, Eureca vzw, GlaxoSmithKline and Teva outside the submitted work. Conflict of interest: N. Krug has nothing to disclose. Conflict of interest: P. Montuschi has nothing to disclose. Conflict of interest: J. Musial has nothing to disclose. Conflict of interest: E. Niżankowska-Mogilnicka has nothing to disclose. Conflict of interest: H.K. Olsson reports other support from AstraZeneca outside the submitted work. Conflict of interest: A. Papi reports grants, personal fees, non-financial support and others from GlaxoSmithKline, AstraZeneca, Boehringer Ingelheim, Chiesi, Teva, Mundipharma, Zambon, Novartis, Menarini, Sanofi/Regeneron, Roche, Fondazione Salvatore Maugeri, Chiesi and Edmond pharma outside the submitted work. Conflict of interest: K.F. Rabe reports grants and personal fees from AstraZeneca, Boehringer Ingelheim, Sanofi Aventis, MERCK SHARP & DOHME, Novartis, Orion Cooperation, Berlin Chemie, Roche, Chiesi and grants for research from the Ministry of Education and Science, Germany. Conflict of interest: T. Sandström has nothing to disclose. Conflict of interest: D.E. Shaw reports personal fees and non-financial support from GlaxoSmithKline, Novartis and AstraZeneca outside the submitted work. Conflict of interest: N.M. Siafakas has nothing to disclose. Conflict of interest: M. Uhlen has nothing to disclose. Conflict of interest: J.H. Riley is an employee and shareholder in GlaxoSmithKline. Conflict of interest: S. Bates is an employee and shareholder in GlaxoSmithKline. Conflict of interest: R.J.M. Middelveld reports grants from the Swedish Strategic Research Foundation, AstraZeneca, the Swedish Heart Lung Foundation and the Swedish Asthma and Allergy Association outside the submitted work. Conflict of interest: C.E. Wheelock has nothing to disclose. Conflict of interest: K.F. Chung reports grants and personal fees from GlaxoSmithKline, AstraZeneca, Novartis, Merck, Boehringer Ingelheim, Roche and Shionogi outside the submitted work. Conflict of interest: I.M. Adcock has nothing to disclose. Conflict of interest: P.J. Sterk reports grants to Amsterdam UMC from the public private Innovative Medicines Initiative (IMI) covered by the European Union (EU) and the European Federation of Pharmaceutical Industries and Associations (EFPIA) during the conduct of the study. Conflict of interest: R. Djukanovic reports receiving fees for lectures at symposia organised by Novartis, GlaxoSmithKline, AstraZeneca and Teva, consultation fees from Teva and Novartis; he is a co-founder and current consultant and has shares in Synairgen. Conflict of interest: P. Nilsson has nothing to disclose. Conflict of interest: S-E. Dahlén reports personal fees from AstraZeneca, Cayman Chemicals, GlaxoSmithKline, Novartis, Regeneron, Sanofi and Teva outside the submitted work. Conflict of interest: A. James has nothing to disclose., (Copyright ©The authors 2022.)

There is a paucity of biomarkers for chronic obstructive pulmonary disease (COPD). Metabolomics were applied to a defined COPD patient cohort from the ECLIPSE study (Evaluation of COPD Longitudinally to Identify Predictive Surrogate End-points). Results were correlated with accepted biomarkers for the disease. Baseline control serum (n=66) and Global Initiative for Chronic Obstructive Lung Disease (GOLD) stage II (n=70), III (n=64) and IV (n=44) COPD patients were analysed by proton nuclear magnetic resonance ((1)H NMR). Liquid chromatography with tandem mass spectrometry (LC-MS/MS) was used to confirm amino acid changes detected by (1)H NMR. Data were correlated with body composition, emphysema and systemic inflammation. (1)H NMR identified decreased lipoproteins, N,N-dimethylglycine, and increased glutamine, phenylalanine, 3-methylhistidine and ketone bodies in COPD patients with decreased branched-chain amino acids (BCAAs) observed in GOLD stage IV patients. BCAAs, their degradation products, 3-methylhistidine, ketone bodies, and triglycerides were correlated negatively with cachexia and positively with systemic inflammation. Emphysema patients also displayed decreased serum creatine, glycine and N,N-dimethylglycine. LC-MS/MS confirmed (1)H NMR findings relating to BCAAs, glutamine and 3-methylhistidine in GOLD stage IV patients. NMR-based metabolomics characterised COPD patients based on systemic effects and lung function parameters. Increased protein turnover occurred in all COPD patients with increased protein degradation in individuals with emphysema and cachexia.