Thiazolidinone CFTR inhibitor identified by high-throughput screening blocks cholera toxin–induced intestinal fluid secretion

Ma, Tao; Thiagarajah, Jay R.; Ye, Hong; Sonawane, N.D.; Folli, Chiara; Galietta, Luis J. V.; Verkman, A. S.

doi:10.1172/jci200216112

Cited by 181 publications

(250 citation statements)

References 17 publications

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“…Except for the efficacy of NPPB, the pharmacological profile for pCFTR is comparable to that for CFTR from other species. DIDS is known not to inhibit CFTR from the external surface (Schultz et al, 1999) and, using the same method used in the present study, similar IC 50 values for human CFTR were obtained for GlyH101 (Muanprasat et al, 2004) and glibenclamide (Ma et al, 2002). The IC 50 values reported for the inhibition of CFTR by NPPB are quite variable, perhaps reflecting the voltage dependence of this block (Hwang and Sheppard, 1999).…”

Section: Discussionsupporting

confidence: 79%

The distribution and expression of CFTR restricts electrogenic anion secretion to the ileum of the brushtail possum, Trichosurus vulpecula

Gill

Bartolo

Demmers

et al. 2011

Journal of Experimental Biology

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SUMMARYIn eutherian mammals, fluid secretion is essential for intestinal function. This is driven by electrogenic Cl -secretion, which involves a NaK2Cl cotransporter (NKCC1) in the enterocyte basolateral membrane and the cystic fibrosis transmembrane conductance regulator (CFTR) in the apical membrane. However, in the possum ileum, NKCC1 expression is low and secretagogues stimulate electrogenic HCO 3 -secretion driven by a basolateral NaHCO 3 cotransporter (pNBCe1). Here we investigated whether electrogenic anion secretion occurs in possum duodenum and jejunum and determined the role of CFTR in possum intestinal anion secretion. Prostaglandin E 2 (PGE 2 ) and forskolin stimulated a large increase in ileal short-circuit current (I sc ), consistent with electrogenic HCO 3 -secretion, but had little effect on the duodenal and jejunal I sc . Furthermore, 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) and N-(2-naphthalenyl)-[(3,5-dibromo-2,4-dihydroxyphenyl)methylene]glycine hydrazide (GlyH101) inhibited cloned possum CFTR in cultured cells and the PGE 2 -stimulated ileal I sc , implicating CFTR in ileal HCO 3 -secretion. Consistent with this, CFTR is expressed in the apical membrane of ileal crypt and lower villous cells, which also express pNBCe1 in the basolateral membrane. In contrast, duodenal and jejunal CFTR expression is low relative to the ileum. Jejunal pNBCe1 expression is also low, whereas duodenal and ileal pNBCe1 expression are comparable. All regions have low NKCC1 expression. These results indicate that cAMP-dependent electrogenic Cl -secretion does not occur in the possum small intestine because of the absence of CFTR and NKCC1. Furthermore, CFTR functions as the apical anion conductance associated with HCO 3 -secretion and its distribution limits electrogenic HCO 3 -secretion to the ileum.

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Section: Discussionsupporting

confidence: 79%

The distribution and expression of CFTR restricts electrogenic anion secretion to the ileum of the brushtail possum, Trichosurus vulpecula

Gill

Bartolo

Demmers

et al. 2011

Journal of Experimental Biology

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“…To elucidate whether the CFTR channel, a plasma membrane Cl Ϫ channel, is involved in fluid accumulation caused by hemolysin, the effects of H-89, a cAMP-dependent protein kinase A (PKA) inhibitor (7,27), and glibenclamide, a selective CFTR inhibitor (34,35), were examined. For the control experiment, CT, which causes diarrhea through the activation of CFTR (18,23), was used. H-89 and glibenclamide dose-dependently inhibited the fluid accumulation induced by hemolysin and CT (Fig.…”

Section: Resultsmentioning

confidence: 99%

Fluid Secretion Caused by Aerolysin-Like Hemolysin of Aeromonas sobria in the Intestines Is Due to Stimulation of Production of Prostaglandin E ₂ via Cyclooxygenase 2 by Intestinal Cells

et al. 2008

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To clarify the mechanisms of diarrheal disease induced by Aeromonas sobria, we examined whether prostaglandin E 2 (PGE 2 ) was involved in the intestinal secretory action of A. sobria hemolysin by use of a mouse intestinal loop model. The amount of PGE 2 in jejunal fluid and the fluid accumulation ratio were directly related to the dose of hemolysin. The increase over time in the level of PGE 2 was similar to that of the accumulated fluid. In addition, hemolysin-induced fluid secretion and PGE 2 synthesis were inhibited by the selective cyclooxygenase 2 (COX-2) inhibitor NS-398 but not the COX-1 inhibitor SC-560. Western blot analysis revealed that hemolysin increased the COX-2 protein levels but reduced the COX-1 protein levels in mouse intestinal mucosa in vivo. These results suggest that PGE 2 functions as an important mediator of diarrhea caused by hemolysin and that PGE 2 is produced primarily through a COX-2-dependent mechanism. Subsequently, we examined the relationship between PGE 2 , cyclic AMP (cAMP), and cystic fibrosis transmembrane conductance regulator (CFTR) Cl ؊ channels in mouse intestinal mucosa exposed to hemolysin. Hemolysin increased the levels of cAMP in the intestinal mucosa. NS-398 inhibited the increase in cAMP production, but SC-560 did not. In addition, H-89, a cAMP-dependent protein kinase A (PKA) inhibitor, and glibenclamide, a CFTR inhibitor, inhibited fluid accumulation. Taken together, these results indicate that hemolysin activates PGE 2 production via COX-2 and that PGE 2 stimulates cAMP production. cAMP then activates PKA, which in turn stimulates CFTR Cl ؊ channels and finally leads to fluid accumulation in the intestines.

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“…High-throughput screening was performed using a semiautomated screening platform (Beckman Coulter, Fullerton, CA, USA) configured as described elsewhere (37). FRT cells expressing human pendrin and EYFP-HIF were plated in 96-well black-walled, clear-bottom tissue culture plates (Corning, Corning, NY, USA) at a density of 20,000 cells/well, and cultured to confluence over 48 h. For screening, plates were washed twice with PBS (in mM: 137 NaCl, 2.7 KCl, 1.1 KH 2 PO 4 , 8.1 Na 2 HPO 4 , 0.9 CaCl 2 , 0.5 MgCl 2 ) before addition of 100 ml PBS containing test compounds.…”

Section: Screening Proceduresmentioning

confidence: 99%

Inhibitors of pendrin anion exchange identified in a small molecule screen increase airway surface liquid volume in cystic fibrosis

Haggie

Phuan²,

Tan³

et al. 2016

FASEB j.

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Pendrin (SLC26A4) is a Cl 2 /anion exchanger expressed in the epithelium of inflamed airways where it is thought to facilitate Cl 2 absorption and HCO 3 2 secretion. Studies using pendrin knockout mice and airway epithelial cells from hearing-impaired subjects with pendrin loss of function suggest involvement of pendrin in inflammatory lung diseases, including cystic fibrosis (CF), perhaps by regulation of airway surface liquid (ASL) volume. Here we identified small-molecule pendrin inhibitors and demonstrated their efficacy in increasing ASL volume. A cell-based, functional high-throughput screen of ∼36,000 synthetic small molecules produced 3 chemical classes of inhibitors of human pendrin. After structure-activity studies, tetrahydropyrazolopyridine and pyrazolothiophenesulfonamide compounds reversibly inhibited pendrin-facilitated Cl 2 exchange with SCN 2 , I 2 , NO 3 2 , and HCO 3 2 with drug concentration causing 50% inhibition down to ∼2.5 mM. In well-differentiated primary cultures of human airway epithelial cells from non-CF and CF subjects, treatment with IL-13, which causes inflammation with strong pendrin up-regulation, strongly increased Cl 2 /HCO 3 2 exchange and the increase was blocked by pendrin inhibition. Pendrin inhibition significantly increased ASL depth (by ∼8 mm) in IL-13-treated non-CF and CF cells but not in untreated cells. These studies implicate the involvement of pendrin-facilitated Cl 2 / HCO 3 2 in the regulation of ASL volume and suggest the utility of pendrin inhibitors in inflammatory lung diseases, including CF

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Thiazolidinone CFTR inhibitor identified by high-throughput screening blocks cholera toxin–induced intestinal fluid secretion

Cited by 181 publications

References 17 publications

The distribution and expression of CFTR restricts electrogenic anion secretion to the ileum of the brushtail possum, Trichosurus vulpecula

The distribution and expression of CFTR restricts electrogenic anion secretion to the ileum of the brushtail possum, Trichosurus vulpecula

Fluid Secretion Caused by Aerolysin-Like Hemolysin of Aeromonas sobria in the Intestines Is Due to Stimulation of Production of Prostaglandin E ₂ via Cyclooxygenase 2 by Intestinal Cells

Inhibitors of pendrin anion exchange identified in a small molecule screen increase airway surface liquid volume in cystic fibrosis

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Thiazolidinone CFTR inhibitor identified by high-throughput screening blocks cholera toxin–induced intestinal fluid secretion

Cited by 181 publications

References 17 publications

The distribution and expression of CFTR restricts electrogenic anion secretion to the ileum of the brushtail possum, Trichosurus vulpecula

The distribution and expression of CFTR restricts electrogenic anion secretion to the ileum of the brushtail possum, Trichosurus vulpecula

Fluid Secretion Caused by Aerolysin-Like Hemolysin of Aeromonas sobria in the Intestines Is Due to Stimulation of Production of Prostaglandin E 2 via Cyclooxygenase 2 by Intestinal Cells

Inhibitors of pendrin anion exchange identified in a small molecule screen increase airway surface liquid volume in cystic fibrosis

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Fluid Secretion Caused by Aerolysin-Like Hemolysin of Aeromonas sobria in the Intestines Is Due to Stimulation of Production of Prostaglandin E ₂ via Cyclooxygenase 2 by Intestinal Cells