TY - JOUR
T1 - Interruption of transmembrane signaling as a novel antisecretory strategy to treat enterotoxigenic diarrhea
AU - Zhang, Wei
AU - Mannan, Ishrat
AU - Schulz, Stephanie
AU - Parkinson, Scott J.
AU - Alekseev, Alexey E.
AU - Gomez, Luis A.
AU - Terzic, Andre
AU - Waldman, Scott A.
PY - 1999
Y1 - 1999
N2 - 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 (2C1ATP) and its cell-permeant precursor, 2- Chloroadenosine (2CIAdo), 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 2CIAdo 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-tozapical 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.
AB - 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 (2C1ATP) and its cell-permeant precursor, 2- Chloroadenosine (2CIAdo), 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 2CIAdo 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-tozapical 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.
KW - 2-substituted adenine nucleotides
KW - CFTR-mediated chloride current
KW - Cyclic GMP
KW - E. coli heat-stable enterotoxin
KW - Intestinal cell water secretion
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U2 - 10.1096/fasebj.13.8.913
DO - 10.1096/fasebj.13.8.913
M3 - Article
C2 - 10224234
AN - SCOPUS:0032896023
SN - 0892-6638
VL - 13
SP - 913
EP - 922
JO - FASEB Journal
JF - FASEB Journal
IS - 8
ER -