Sex-dependent regulation of mucin gene transcription and airway secretion and mechanics following intra-airway IL-13 in mice with conditional loss of club cell Creb1

Mariana Sponchiado; Amy Fagan; Luz Mata; Angelina L Bonilla; Pedro Trevizan-Baú; Sreekala Prabhakaran; Leah R Reznikov

doi:10.3389/fphys.2024.1392443

Sex-dependent regulation of mucin gene transcription and airway secretion and mechanics following intra-airway IL-13 in mice with conditional loss of club cell Creb1

Front Physiol. 2024 Apr 22:15:1392443. doi: 10.3389/fphys.2024.1392443. eCollection 2024.

Authors

Mariana Sponchiado^#¹, Amy Fagan^#¹, Luz Mata¹, Angelina L Bonilla¹, Pedro Trevizan-Baú¹, Sreekala Prabhakaran², Leah R Reznikov¹

Affiliations

¹ Department of Physiological Sciences, University of Florida, Gainesville, FL, United States.
² Department of Pediatrics Pediatric Pulmonary Division, University of Florida, Gainesville, FL, United States.

^# Contributed equally.

Abstract

Introduction: Interleukin 13 (IL-13) is an important effector molecule in allergic asthma. IL-13-mediated mucin hypersecretion requires conversion of secretoglobin-positive club cells into goblet cells through suppression of forkhead box A2 (FOXA2) and induction of SAM pointed domain containing ETS transcription factor (SPDEF). IL-13-mediated mucin hypersecretion may also include modulation of purinergic and muscarinic receptors that control basal and stimulated mucin secretion. We recently found that the transcription factor cAMP response element-binding protein (Creb1) inhibits FOXA2 and modulates mucus secretion in mice. Methods: We tested the hypothesis that loss of club cell Creb1 mitigates the pro-mucin effects of IL-13. We challenged male and female mice with conditional loss of club cell Creb1 and wild type littermates with intra-airway IL-13 or vehicle. We also studied human "club cell-like" NCI-H322 cells. Results: Loss of club cell Creb1 augmented IL-13-mediated increases in mRNA for the gel-forming mucins Muc5ac and Muc5b and prevented IL-13-mediated decreases in muscarinic 3 receptor (M3R) mRNA in male airways. In female airways, loss of club cell Creb1 reduced M3R mRNA and significantly blunted IL-13-mediated increases in purinergic receptor P2Y2 (P2ry2) mRNA but did not impact Muc5ac and Muc5b mRNA. Despite changes in mucins and secretion machinery, goblet cell density following cholinergic stimulation was not impacted by loss of club cell Creb1 in either sex. IL-13 treatment decreased basal airway resistance across sexes in mice with loss of club cell Creb1, whereas loss of club cell Creb1 augmented IL-13-mediated increases in airway elastance in response to methacholine. NCI-H322 cells displayed IL-13 signaling components, including IL-13Rα1 and IL-4Rα. Pharmacologic inhibition of CREB reduced IL-13Rα1 mRNA, whereas recombinant CREB decreased IL-4Rα mRNA. Application of IL-13 to NCI-H322 cells increased concentrations of cAMP in a delayed manner, thus linking IL-13 signaling to CREB signaling. Conclusion: These data highlight sex-specific regulation of club cell Creb1 on IL-13-mediated mucin hypersecretion and airway mechanics.

Keywords: IL-13; airway mechanics; club cell; mucin; sex differences.

Grants and funding

The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. This work was supported in part by the National Institutes of Health (OD026582, HL152101), the Cystic Fibrosis Foundation (REZNIKO20I0, REZNIKO19I0), and the University of Florida McKnight Career Accelerator Award. Angelina Bonilla is funded by the National institutes of Health (R25GM115298).