Inflammatory macrophage memory in nonsteroidal anti-inflammatory drug–exacerbated respiratory disease

doi:10.1016/j.jaci.2020.04.064

Journal of Allergy and Clinical Immunology

Volume 147, Issue 2, February 2021, Pages 587-599

https://doi.org/10.1016/j.jaci.2020.04.064 Get rights and content

Background

Nonsteroidal anti-inflammatory drug–exacerbated respiratory disease (N-ERD) is a chronic inflammatory condition, which is driven by an aberrant arachidonic acid metabolism. Macrophages are major producers of arachidonic acid metabolites and subject to metabolic reprogramming, but they have been neglected in N-ERD.

Objective

This study sought to elucidate a potential metabolic and epigenetic macrophage reprogramming in N-ERD.

Methods

Transcriptional, metabolic, and lipid mediator profiles in macrophages from patients with N-ERD and healthy controls were assessed by RNA sequencing, Seahorse assays, and LC-MS/MS. Metabolites in nasal lining fluid, sputum, and plasma from patients with N-ERD (n = 15) and healthy individuals (n = 10) were quantified by targeted metabolomics analyses. Genome-wide methylomics were deployed to define epigenetic mechanisms of macrophage reprogramming in N-ERD.

Results

This study shows that N-ERD monocytes/macrophages exhibit an overall reduction in DNA methylation, aberrant metabolic profiles, and an increased expression of chemokines, indicative of a persistent proinflammatory activation. Differentially methylated regions in N-ERD macrophages included genes involved in chemokine signaling and acylcarnitine metabolism. Acylcarnitines were increased in macrophages, sputum, nasal lining fluid, and plasma of patients with N-ERD. On inflammatory challenge, N-ERD macrophages produced increased levels of acylcarnitines, proinflammatory arachidonic acid metabolites, cytokines, and chemokines as compared to healthy macrophages.

Conclusions

Together, these findings decipher a proinflammatory metabolic and epigenetic reprogramming of macrophages in N-ERD.

Graphical abstract

Section snippets

Materials

A detailed list of all materials used in this study can be found in this article’s Online Repository available at www.jacionline.org.

Patient characterization

Patients with N-ERD and healthy controls were recruited and classified according to their clinical characteristics at the ear, nose, and throat clinic of the Klinikum rechts der Isar (Munich, Germany) (see Table E1 in this article’s Online Repository at www.jacionline.org for an overview). Malm score²⁵ was determined by nasal endoscopy. Healthy controls had no

Patients with N-ERD exhibit a persistent proinflammatory macrophage activation

MDMs populate the airways, plastically respond to inflammatory stimuli, and govern immune responses during type 2 inflammation.⁴⁰^,⁴¹ To identify a potential macrophage reprogramming in N-ERD, we performed an RNAseq analysis of aMDMs, differentiated from CD14⁺ blood monocytes from patients with N-ERD and healthy individuals in a lung-adapted cytokine milieu (TGF-β1 and GM-CSF).¹⁹ Despite 1 week of in vitro differentiation, 86 downregulated and 19 upregulated genes were identified in N-ERD as

Discussion

Chronic type 2 inflammatory airway diseases including N-ERD remain major unmet clinical needs.²^,⁵⁵ Macrophages are key players in type 2 inflammation,⁴⁰^,⁵⁶ but the metabolic and epigenetic programs that determine macrophage phenotypes and functions in human patients remain largely unknown. The present study uncovers an unprecedented proinflammatory “macrophage memory” in patients suffering from N-ERD, a severe and therapy-resistant form of chronic airway inflammation. A combination of

References (74)

J.P. Rajan et al.
Prevalence of aspirin-exacerbated respiratory disease among asthmatic patients: a meta-analysis of the literature
J Allergy Clin Immunol
(2015)
W.W. Stevens et al.
Clinical characteristics of patients with chronic rhinosinusitis with nasal polyps, asthma, and aspirin-exacerbated respiratory disease
J Allergy Clin Immunol Pract
(2017)
L. Fernández-Bertolín et al.
Deficient glucocorticoid induction of anti-inflammatory genes in nasal polyp fibroblasts of asthmatic patients with and without aspirin intolerance
J Allergy Clin Immunol
(2013)
T.M. Laidlaw et al.
Prostaglandin E2 resistance in granulocytes from patients with aspirin-exacerbated respiratory disease
J Allergy Clin Immunol
(2014)
S.C.-C. Huang et al.
Metabolic reprogramming mediated by the mTORC2-IRF4 signaling axis is essential for macrophage alternative activation
Immunity
(2016)
D.L. Clarke et al.
Dectin-2 sensing of house dust mite is critical for the initiation of airway inflammation
Mucosal Immunol
(2014)
T. Takabayashi et al.
Increased expression of factor XIII-A in patients with chronic rhinosinusitis with nasal polyps
J Allergy Clin Immunol
(2013)
T. Koga et al.
Characterisation of patients with frequent exacerbation of asthma
Respir Med
(2006)
S.A. Gharib et al.
Transcriptional and functional diversity of human macrophage repolarization
J Allergy Clin Immunol
(2019)
B.L.K. Chawes et al.
A novel method for assessing unchallenged levels of mediators in nasal epithelial lining fluid
J Allergy Clin Immunol
(2010)

U.M. Zissler et al.

Biomatrix for upper and lower airway biomarkers in patients with allergic asthma

J Allergy Clin Immunol

(2018)

L. Rossi et al.

Gpr171, a putative P2Y-like receptor, negatively regulates myeloid differentiation in murine hematopoietic progenitors

Exp Hematol

(2013)

K. Dietz et al.

Age dictates a steroid-resistant cascade of Wnt5a, transglutaminase 2, and leukotrienes in inflamed airways

J Allergy Clin Immunol

(2017)

T. Erdogan et al.

Comorbid diseases in aspirin-exacerbated respiratory disease, and asthma

Allergol Immunopathol (Madr)

(2015)

S.A. Shore

Obesity and asthma: possible mechanisms

J Allergy Clin Immunol

(2008)

S.A. Shore et al.

Adiponectin attenuates allergen-induced airway inflammation and hyperresponsiveness in mice

J Allergy Clin Immunol

(2006)

X. Cheng et al.

Adiponectin induces pro-inflammatory programs in human macrophages and CD4+ T cells

J Biol Chem

(2012)

M.C. Peters et al.

Refractory airway type 2 inflammation in a large subgroup of asthmatic patients treated with inhaled corticosteroids

J Allergy Clin Immunol

(2019)

X. Yu et al.

The cytokine TGF-β promotes the development and homeostasis of alveolar macrophages

Immunity

(2017)

I. Mitroulis et al.

Modulation of myelopoiesis progenitors is an integral component of trained immunity

Cell

(2018)

L.E. Donnelly et al.

Defective phagocytosis in airways disease

Chest

(2012)

A. Saradna et al.

Macrophage polarization and allergic asthma

Transl Res

(2018)

K.J. Staples et al.

Phenotypic characterisation of lung macrophages in asthma: over-expression of CCL17

J Allergy Clin Immunol

(2012)

W.J. Fokkens et al.

EPOS 2012: European position paper on rhinosinusitis and nasal polyps 2012: a summary for otorhinolaryngologists

Rhinol J

(2012)

M.L. Kowalski et al.

Diagnosis and management of NSAID -Exacerbated Respiratory Disease (N-ERD)—a EAACI position paper

Allergy

(2019)

M. Samter et al.

Intolerance to aspirin: clinical studies and consideration of its pathogenesis

Ann Intern Med

(1968)

T.M. Laidlaw et al.

Aspirin-exacerbated respiratory disease—new prime suspects

N Engl J Med

(2016)

K.E. Hulse et al.

Pathogenesis of nasal polyposis

Clin Exp Allergy

(2015)

S.-E. Dahlén et al.

Improvement of aspirin-intolerant asthma by montelukast, a leukotriene antagonist: a randomized, double-blind, placebo-controlled trial

Am J Respir Crit Care Med

(2002)

F. Gaber et al.

Increased levels of cysteinyl-leukotrienes in saliva, induced sputum, urine and blood from patients with aspirin-intolerant asthma

Thorax

(2008)

L. Mastalerz et al.

Induced sputum eicosanoids during aspirin bronchial challenge of asthmatic patients with aspirin hypersensitivity

Allergy

(2014)

J. Eriksson et al.

Aspirin-intolerant asthma in the population: prevalence and important determinants

Clin Exp Allergy

(2015)

H.K.T. Trinh et al.

Exploration of the sphingolipid metabolite, sphingosine-1-phosphate and sphingosine, as novel biomarkers for aspirin-exacerbated respiratory disease

Sci Rep

(2016)

A.A. Al-Khami et al.

Fueling the mechanisms of asthma: increased fatty acid oxidation in inflammatory immune cells may represent a novel therapeutic target

Clin Exp Allergy

(2017)

C. Draijer et al.

PGE2-treated macrophages inhibit development of allergic lung inflammation in mice

J Leukoc Biol

(2016)

F.D.R. Henkel et al.

House dust mite drives proinflammatory eicosanoid reprogramming and macrophage effector functions

Allergy

(2019)

E.M. Varga et al.

Inflammatory cell populations and cytokine mRNA expression in the nasal mucosa in aspirin-sensitive rhinitis

Eur Respir J

(1999)

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Macrophages represent key regulators of lung homeostasis and immunity, and they govern airway inflammation by producing eicosanoids and chemokines.15,27 We recently described stable differences in gene expression and metabolite profiles in macrophages from patients with nonsteroidal anti-inflammatory drug–exacerbated respiratory disease,26 a nonallergic chronic T2 inflammatory condition. To study a potential macrophage memory in allergic asthma, we generated macrophages (aMDM) from monocytes of HDM-allergic or healthy donors (Table I) (Fig E1, A).
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: This study was supported by the Else Kröner-Fresenius-Stiftung (grant 2015_A195), the German Research Foundation (FOR2599, ES 471/3-1), the Fritz Thyssen Stiftung (grant Az. 10.17.2.017MN), and a Helmholtz Young Investigator grant (VH-NG-1331) to JEvB. CSW receives grant support by the German Center for Lung Research (82DZL00302). This study was supported in part by a grant from the German Federal Ministry of Education and Research to the German Center Diabetes Research.

: Disclosure of potential conflict of interest: C. B. Schmidt-Weber received grant support from Allergopharma, PLS Design, as well as Zeller AG; and received speaker honoraria from Allergopharma. The rest of the authors declare that they have no relevant conflicts of interest.

^∗: Member of the German Center of Lung Research.

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Asthma and lower airway diseaseInflammatory macrophage memory in nonsteroidal anti-inflammatory drug–exacerbated respiratory disease

Background

Objective

Methods

Results

Conclusions

Graphical abstract

Section snippets

Materials

Patient characterization

Patients with N-ERD exhibit a persistent proinflammatory macrophage activation

Discussion

J Allergy Clin Immunol

J Allergy Clin Immunol Pract

J Allergy Clin Immunol

J Allergy Clin Immunol

Immunity

Mucosal Immunol

J Allergy Clin Immunol

Respir Med

J Allergy Clin Immunol

J Allergy Clin Immunol

J Allergy Clin Immunol

Exp Hematol

J Allergy Clin Immunol

Allergol Immunopathol (Madr)

J Allergy Clin Immunol

J Allergy Clin Immunol

J Biol Chem

J Allergy Clin Immunol

Immunity

Cell

Chest

Transl Res

J Allergy Clin Immunol

EPOS 2012: European position paper on rhinosinusitis and nasal polyps 2012: a summary for otorhinolaryngologists

Rhinol J

Diagnosis and management of NSAID -Exacerbated Respiratory Disease (N-ERD)—a EAACI position paper

Allergy

Intolerance to aspirin: clinical studies and consideration of its pathogenesis

Ann Intern Med

Aspirin-exacerbated respiratory disease—new prime suspects

N Engl J Med

Pathogenesis of nasal polyposis

Clin Exp Allergy

Improvement of aspirin-intolerant asthma by montelukast, a leukotriene antagonist: a randomized, double-blind, placebo-controlled trial

Am J Respir Crit Care Med

Increased levels of cysteinyl-leukotrienes in saliva, induced sputum, urine and blood from patients with aspirin-intolerant asthma

Thorax

Induced sputum eicosanoids during aspirin bronchial challenge of asthmatic patients with aspirin hypersensitivity

Allergy

Aspirin-intolerant asthma in the population: prevalence and important determinants

Clin Exp Allergy

Exploration of the sphingolipid metabolite, sphingosine-1-phosphate and sphingosine, as novel biomarkers for aspirin-exacerbated respiratory disease

Sci Rep

Fueling the mechanisms of asthma: increased fatty acid oxidation in inflammatory immune cells may represent a novel therapeutic target

Clin Exp Allergy

PGE2-treated macrophages inhibit development of allergic lung inflammation in mice

J Leukoc Biol

House dust mite drives proinflammatory eicosanoid reprogramming and macrophage effector functions

Allergy

Inflammatory cell populations and cytokine mRNA expression in the nasal mucosa in aspirin-sensitive rhinitis

Eur Respir J

Asthma and lower airway disease
Inflammatory macrophage memory in nonsteroidal anti-inflammatory drug–exacerbated respiratory disease