Analysis of Lung Gene Expression Reveals a Role for Cl- Channels in Diisocyanate-induced Airway Eosinophilia in a Mouse Model of Asthma Pathology

Am J Respir Cell Mol Biol. 2020 Jul;63(1):25-35. doi: 10.1165/rcmb.2019-0400OC.

Abstract

Diisocyanates are well-recognized causes of asthma. However, sensitized workers frequently lack diisocyanate-specific IgE, which complicates diagnosis and suggests the disease involves IgE-independent mechanisms. We used a mouse model of methylene diphenyl diisocyanate (MDI) asthma to identify biological pathways that may contribute to asthma pathogenesis. MDI sensitization and respiratory tract exposure were performed in Balb/c, transgenic B-cell (e.g., IgE)-deficient mice and a genetic background (C57BL/6)-matched strain. Eosinophils in airway fluid were quantitated by flow cytometry. Lung tissue gene expression was assessed using whole-genome mRNA microarrays. Informatic software was used to identify biological pathways affected by respiratory tract exposure and potential targets for disease intervention. Airway eosinophilia and changes (>1.5-fold; P value < 0.05) in expression of 192 genes occurred in all three mouse strains tested, with enrichment in chemokines and a pattern associated with alternatively activated monocytes/macrophages. CLCA1 (calcium-activated chloride channel regulator 1) was the most upregulated gene transcript (>100-fold) in all exposed mouse lungs versus controls, followed closely by SLC26A4, another transcript involved in Cl- conductance. Crofelemer, a U.S. Food and Drug Administration-approved Cl- channel inhibitor, reduced MDI exposure induction of airway eosinophilia, mucus, CLCA1, and other asthma-associated gene transcripts. Expression changes in a core set of genes occurs independent of IgE in a mouse model of chemical-induced airway eosinophilia. In addition to chemokines and alternatively activated monocytes/macrophages, the data suggest a crucial role for Cl- channels in diisocyanate asthma pathology and as a possible target for intervention.

Keywords: asthma; channel; chloride; crofelemer; diisocyanate.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Asthma / metabolism*
  • Asthma / pathology*
  • Bronchoalveolar Lavage Fluid
  • Chloride Channels / metabolism*
  • Disease Models, Animal
  • Eosinophilia / chemically induced
  • Eosinophilia / metabolism*
  • Eosinophils / metabolism*
  • Eosinophils / pathology
  • Gene Expression / drug effects
  • Gene Expression / physiology*
  • Immunoglobulin E / metabolism
  • Isocyanates / pharmacology
  • Lung / drug effects
  • Lung / metabolism*
  • Lung / pathology
  • Macrophages / drug effects
  • Macrophages / metabolism
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Monocytes / drug effects
  • Monocytes / metabolism

Substances

  • Chloride Channels
  • Isocyanates
  • Immunoglobulin E
  • 4,4'-diphenylmethane diisocyanate