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Cited by 53Ion Transport and Channel RegulationCystic Fibrosis Research AdvancesDrug Transport and Resistance Mechanisms

cAMP Stimulates SLC26A3 Activity in Human Colon by a CFTR-Dependent Mechanism That Does Not Require CFTR Activity

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Tse, Chung-Ming, Yin, Jianyi, Singh, Varsha, Sarker, Rafiquel, Lin, Ruxian, Verkman, Alan S., Turner, Jerrold R., Donowitz, Mark·2019·Cellular and Molecular Gastroenterology and Hepatology
DOI 10.1016/j.jcmgh.2019.01.002PMC PMC6438990

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Abstract

Abstract Background & Aims DRA (SLC26A3) is an electroneutral Cl - /HCO 3 - exchanger that is present in the apical domain of multiple intestinal segments. An area that has continued to be poorly understood is related to DRA regulation in acute cAMP-related diarrheas, in which DRA appears to be both inhibited as part of NaCl absorption and stimulated to contribute to increased HCO 3 - secretion. Different cell models expressing DRA have shown that cAMP inhibits, stimulates or does not affect its activity. Methods This study reevaluated cAMP regulation of DRA using new “tools” including a successful knockout cell model, a specific DRA inhibitor (DRA inh -A250), specific antibodies, and a transport assay that did not rely on non-specific inhibitors. The studies compared DRA regulation in colonoids made from normal human colon with regulation in the colon cancer cell line, Caco-2. Results DRA is an apical protein in human proximal colon, differentiated colonoid monolayers and Caco-2 cells. It is glycosylated and appears as two bands. cAMp(forskolin) acutely stimulated DRA activity in human colonoids and Caco-2 cells. In these cells, DRA is the predominant apical Cl - /HCO 3 - exchanger and is inhibited by DRA inh -A250 with IC 50 of 0.5 μmol/L and 0.2 µ mol/L, respectively. However, there was no effect of cAMP in HEK293/DRA cells that lacked CFTR. When CFTR was expressed in HEK293/DRA cells, cAMP also stimulated DRA activity. In all cases, cAMP stimulation of DRA was not inhibited by CFTR inh -172. Conclusions DRA is acutely stimulated by cAMP by a process that is CFTR-dependent but appears to be one of multiple regulatory effects of CFTR that does not require CFTR activity.

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  • Human intestinal organoid-derived monolayers have been successfully used to study host-pathogen interactions, epithelial barrier function, and various transport processes including secretion of hormones, growth factors, and ions, and absorption of glucose.

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    From Regulation of Enterocyte Brush Border Membrane Primary Na-Absorptive Transporters in Human Intestinal Organoid-Derived Monolayers (2024) · cites this paper as [15]

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Authors
Tse, C.-M., Yin, J., Singh, V., Sarker, R., Lin, R., Verkman, A. S., Turner, J. R., & Donowitz, M.
Affiliations
  • Johns Hopkins University (US)
  • University of California, San Francisco (US)
  • Brigham and Women's Hospital (US)
Corresponding author
Donowitz, Mark
ORCID
Venue
Cellular and Molecular Gastroenterology and Hepatology
Publisher
Elsevier BV
Published
2019
Volume
7
Issue
3
Pages
641-653
ISSN
2352-345X (ISSN-L)
DOI
10.1016/j.jcmgh.2019.01.002
PMC
PMC6438990
OpenAlex
W2885038155
Cited by
53
References
1
Citations by year
2026: 2 · 2025: 8 · 2024: 8 · 2023: 6 · 2022: 10 · 2021: 6 · 2020: 8 · 2019: 4 · 2018: 1
FWCI
2.32
Type
journal article
Language
en
Open access
Licence
Locations
  • Cellular and Molecular Gastroenterology and Hepatology (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • eScholarship (California Digital Library) (submittedVersion)
  • eScholarship (California Digital Library) (submittedVersion, cc-by-nc-nd)
Funders
National Institutes of Health (R01-DK-26523, R01-DK-61765, P01-DK-072084, R24-DK-64388, U18-TR000552, UH3-TR00003, UO1-DK-10316, P30-DK-89502)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion Transport and Channel Regulation
  • Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Cystic Fibrosis Research Advances
  • Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
Keywords
SLC26A3, CFTR, cyclic AMP, DRA, colon, colonoids, Caco-2 cells, epithelial ion transport, colon cancer
MeSH terms
Sulfate Transporters, Cyclic AMP, Cell Differentiation, Colon, Colforsin, Humans, Organoids, Reproducibility of Results, Ion Transport, Caco-2 Cells, Cystic Fibrosis Transmembrane Conductance Regulator, Chloride-Bicarbonate Antiporters, HEK293 Cells

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