The Anion Channel TMEM16a/Ano1 Modulates CFTR Activity, but Does Not Function as an Apical Anion Channel in Colonic Epithelium from Cystic Fibrosis Patients and Healthy Individuals.

Abstract

Studies in human colonic cell lines and murine intestine suggest the presence of a Ca2+-activated anion channel, presumably TMEM16a. Is there a potential for fluid secretion in patients with severe cystic fibrosis transmembrane conductance regulator (CFTR) mutations by activating this alternative pathway? Two-dimensional nondifferentiated colonoid-myofibroblast cocultures resembling transit amplifying/progenitor (TA/PE) cells, as well as differentiated monolayer (DM) cultures resembling near-surface cells, were established from both healthy controls (HLs) and patients with severe functional defects in the CFTR gene (PwCF). F508del mutant and CFTR knockout (null) mice ileal and colonic mucosa was also studied. HL TA/PE monolayers displayed a robust short-circuit current response (ΔIeq) to UTP (100 µM), forskolin (Fsk, 10 µM) and carbachol (CCH, 100 µM), while ΔIeq was much smaller in differentiated monolayers. The selective TMEM16a inhibitor Ani9 (up to 30 µM) did not alter the response to luminal UTP, significantly decreased Fsk-induced ΔIeq, and significantly increased CCH-induced ΔIeq in HL TA/PE colonoid monolayers. The PwCF TA/PE and the PwCF differentiated monolayers displayed negligible agonist-induced ΔIeq, without a significant effect of Ani9. When TMEM16a was localized in intracellular structures, a staining in the apical membrane was not detected. TMEM16a is highly expressed in human colonoid monolayers resembling transit amplifying cells of the colonic cryptal neck zone, from both HL and PwCF. While it may play a role in modulating agonist-induced CFTR-mediated anion currents, it is not localized in the apical membrane, and it has no function as an apical anion channel in cystic fibrosis (CF) and healthy human colonic epithelium.

• Authors: Azam Salari, Renjie Xiu, Mahdi Amiri, Sophia Theres Pallenberg, Rainer Schreiber, Anna-Maria Dittrich, Burkhard Tümmler, Karl Kunzelmann, Ursula Seidler
• Journal: International journal of molecular sciences (Int J Mol Sci) Vol. 24 Issue 18 (Sep 18 2023) ISSN: 1422-0067 [Electronic] Switzerland
• PMID: 37762516 (Publication Type: Journal Article)

Chemical References

• Anions
• CFTR protein, human
• Cystic Fibrosis Transmembrane Conductance Regulator
• Uridine Triphosphate
• ANO1 protein, human

Topics

• Animals
• Humans
• Mice
• Anions
• Cystic Fibrosis (genetics)
• Cystic Fibrosis Transmembrane Conductance Regulator (genetics)
• Epithelium
• Uridine Triphosphate