title: Interruption of transmembrane signaling as a novel antisecretory strategy to treat enterotoxigenic diarrhea.
abstract: Bacteria that produce heat-stable enterotoxins (STs), a leading cause of secretory diarrhea, are a major cause of morbidity and mortality worldwide. ST stimulates guanylyl cyclase C (GCC) and accumulation of intracellular cyclic GMP ([cGMP]i), which opens the cystic fibrosis transmembrane conductance regulator (CFTR)-related chloride channel, triggering intestinal secretion. Although the signaling cascade mediating ST-induced diarrhea is well characterized, antisecretory therapy targeting this pathway has not been developed. 2-ChloroATP (2ClATP) and its cell-permeant precursor, 2-chloroadenosine (2ClAdo), disrupt ST-dependent signaling in intestinal cells. However, whether the ability to disrupt guanylyl cyclase signaling translates into effective antisecretory therapy remains untested. In this study, the efficacy of 2ClAdo to prevent ST-induced water secretion by human intestinal cells was examined. In Caco-2 human intestinal cells, ST increased [cGMP]i, induced a chloride current, and stimulated net basolateral-to-apical water secretion. This effect on chloride current and water secretion was mimicked by the cell-permeant analog of cGMP, 8-bromo-cGMP. Treatment of Caco-2 cells with 2ClAdo prevented ST-induced increases in [cGMP]i, chloride current and water secretion. Inhibition of the downstream consequences of ST-GCC interaction reflects proximal disruption of cGMP production because 8-bromo-cGMP stimulated chloride current and water secretion in 2ClAdo-treated cells. Thus, this study demonstrates that disruption of guanylyl cyclase signaling is an effective strategy for antisecretory therapy and provides the basis for developing mechanism-based treatments for enterotoxigenic diarrhea.
authors.affiliation: Division of Clinical Pharmacology, Departments of Medicine and Biochemistry and Molecular Pharmacology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
authors.forename: W
authors.initials: W
authors.lastname: Zhang
authors.forename: I
authors.initials: I
authors.lastname: Mannan
authors.forename: S
authors.initials: S
authors.lastname: Schulz
authors.forename: S J
authors.initials: SJ
authors.lastname: Parkinson
authors.forename: A E
authors.initials: AE
authors.lastname: Alekseev
authors.forename: L A
authors.initials: LA
authors.lastname: Gomez
authors.forename: A
authors.initials: A
authors.lastname: Terzic
authors.forename: S A
authors.initials: SA
authors.lastname: Waldman
mesh_terms: D015762:2-Chloroadenosine / Q000494:pharmacology; D001427:Bacterial Toxins / Q000633:toxicity*; D001693:Biological Transport, Active / Q000187:drug effects; D018938:Caco-2 Cells; D002454:Cell Differentiation; D002712:Chlorides / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000187:drug effects / Q000378:metabolism; D003967:Diarrhea / Q000188:drug therapy* / Q000209:etiology* / Q000503:physiopathology; D004768:Enterotoxins / Q000633:toxicity*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D029968:Escherichia coli Proteins; D006162:Guanylate Cyclase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D017136:Ion Transport / Q000187:drug effects; D010766:Phosphorylation; D011355:Prodrugs / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology; D014867:Water / Q000378:metabolism
chemical_list: D001427:Bacterial Toxins; C505032:CFTR protein, human; D002712:Chlorides; D004768:Enterotoxins; D004791:Enzyme Inhibitors; D029968:Escherichia coli Proteins; D011355:Prodrugs; C034151:heat stable toxin (E coli); D014867:Water; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D015762:2-Chloroadenosine; D006162:Guanylate Cyclase