Supplemental Tables for CFTR-Independent Prostaglandin E2-Stimulated Chloride Secretion in Human Airway
Year: 2025
Type: supplementary-materials
Abstract: Background: Prostaglandin E2 (PGE2) is a potent stimulator of airway epithelial Cl- secretion. PGE2 can stimulate CFTR-independent Cl- secretion from Calu-3 submucosal gland cells, whereas human bronchial epithelial (HBE) cells require CFTR. Aim: Determine the mechanism(s) driving CFTR-independent PGE2-stimulated Cl- secretion in Calu-3 cells. Methods: Short-circuit current (Isc) was measured in Calu-3, HBE, and duodenal enteroids in Ussing Chambers. mRNA expression and intracellular Ca2+ (Ca2+i) was determined by qPCR and Fura-Red imaging, respectively. Results: In Calu-3 and HBE cells, PGE2-stimulated Isc was reduced by bilateral and basolateral-only removal of extracellular Ca2+ (Ca2+e), but not by inhibition of protein kinase A (PKA), inositol 1,4,5-triphosphate (IP3), or Ca2+i stores. Duodenal enteroids utilized PKA, IP3, Ca2+i and Ca2+e. EP receptor mRNA expression and functional measurements indicated EP4 receptor dominance in Calu-3 cells. EP4 receptor agonist CAY-10598 (CFTRinh-172, glibenclamide) increased Ca2+i and Isc was driven by Ca2+-activated Cl- secretion. Isc was inhibited by dasatinib, wortmannin, and GSK650394, indicating involvement of Src, phosphoinositol phosphate (PI3K), serum glucocorticoid kinase 1 (SGK1). CFTR-independent CAY-10598-stimulated Isc was mediated by apical Ca2+ release-activated Ca2+ channels (CRAC), P2X receptors, and basolateral TRPV channels. Conclusions: Calu-3 and HBE cells predominantly utilize EP4 receptors and Ca2+e-mediated signaling for PGE2-stimulated Cl- secretion. However, Calu-3 cells leverage apical Ca2+ entry through CRAC and P2X receptors, together with basolateral TRPV activation, Src, PI3K, and SGK1 signaling, for CFTR-independent Cl-secretion. Gaining insights into means to increase CFTR-independent airway Cl- secretion may identify novel therapies to help ameliorate lung diseases with compromised CFTR function.
Source: Figshare
Author: Sellers, Zachary
Language: English
Subfield: Pulmonary and Respiratory Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: green
OpenAlex ID: W6902374720
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