Potent vasorelaxant activity of the TMEM16A inhibitor T16A inh-A01

Alison J. Davis; Jian Shi; Harry A. T. Pritchard; Preet S. Chadha; Normand Leblanc; Georgios Vasilikostas; Zhen Yao; A. S. Verkman; Anthony P. Albert; Iain A. Greenwood

doi:10.1111/j.1476-5381.2012.02199.x

Potent vasorelaxant activity of the TMEM16A inhibitor T16A _inh-A01

Alison J. Davis, Jian Shi, Harry A. T. Pritchard, Preet S. Chadha, Normand Leblanc, Georgios Vasilikostas, Zhen Yao, A. S. Verkman, Anthony P. Albert, Iain A. Greenwood

Norwich Medical School

Research output: Contribution to journal › Article › peer-review

83 Citations (Scopus)

Abstract

Background and Purpose: T16A_inh-A01 is a recently identified inhibitor of the calcium-activated chloride channel TMEM16A. The aim of this study was to test the efficacy of T16A_inh-A01 for inhibition of calcium-activated chloride channels in vascular smooth muscle and consequent effects on vascular tone. Experimental Approach: Single channel and whole cell patch clamp was performed on single smooth muscle cells from rabbit pulmonary artery and mouse thoracic aorta. Isometric tension studies were performed on mouse thoracic aorta and mesenteric artery as well as human abdominal visceral adipose artery. Key Results: In rabbit pulmonary artery myocytes T16A _inh-A01 (1-30 μM) inhibited single calcium (Ca²⁺)- activated chloride (Cl^-) channels and whole cell currents activated by 500 nM free Ca²⁺. Similar effects were observed for single Ca ²⁺-activated Cl^- channels in mouse thoracic aorta, and in both cell types, channel activity was abolished by two antisera raised against TMEM16A but not by a bestrophin antibody. The TMEM16A potentiator, F _act (10 μM), increased single channel and whole cell Ca ²⁺-activated Cl^- currents in rabbit pulmonary arteries. In isometric tension studies, T16A_inh-A01 relaxed mouse thoracic aorta pre-contracted with methoxamine with an IC₅₀ of 1.6 μM and suppressed the methoxamine concentration-effect curve. T16A_inh-A01 did not affect the maximal contraction produced by 60 mM KCl and the relaxant effect of 10 μM T16A_inh-A01 was not altered by incubation of mouse thoracic aorta in a cocktail of potassium (K⁺) channel blockers. T16A_inh-A01 (10 μM) also relaxed human visceral adipose arteries by 88 ± 3%. Conclusions and Implications T16A_inh-A01 blocks calcium-activated chloride channels in vascular smooth muscle cells and relaxes murine and human blood vessels. British Journal of Pharmacology

Original language	English
Pages (from-to)	773-784
Number of pages	12
Journal	British Journal of Pharmacology
Volume	168
Issue number	3
Early online date	5 Sept 2012
DOIs	https://doi.org/10.1111/j.1476-5381.2012.02199.x
Publication status	Published - Feb 2013

Keywords

Ano1
anoctamins
anti-hypertensive
calcium-activated chloride channels
patch clamp
TMEM16A
vascular smooth muscle

Access to Document

10.1111/j.1476-5381.2012.02199.x

Cite this

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title = "Potent vasorelaxant activity of the TMEM16A inhibitor T16A inh-A01",

abstract = "Background and Purpose: T16Ainh-A01 is a recently identified inhibitor of the calcium-activated chloride channel TMEM16A. The aim of this study was to test the efficacy of T16Ainh-A01 for inhibition of calcium-activated chloride channels in vascular smooth muscle and consequent effects on vascular tone. Experimental Approach: Single channel and whole cell patch clamp was performed on single smooth muscle cells from rabbit pulmonary artery and mouse thoracic aorta. Isometric tension studies were performed on mouse thoracic aorta and mesenteric artery as well as human abdominal visceral adipose artery. Key Results: In rabbit pulmonary artery myocytes T16A inh-A01 (1-30 μM) inhibited single calcium (Ca2+)- activated chloride (Cl-) channels and whole cell currents activated by 500 nM free Ca2+. Similar effects were observed for single Ca 2+-activated Cl- channels in mouse thoracic aorta, and in both cell types, channel activity was abolished by two antisera raised against TMEM16A but not by a bestrophin antibody. The TMEM16A potentiator, F act (10 μM), increased single channel and whole cell Ca 2+-activated Cl- currents in rabbit pulmonary arteries. In isometric tension studies, T16Ainh-A01 relaxed mouse thoracic aorta pre-contracted with methoxamine with an IC50 of 1.6 μM and suppressed the methoxamine concentration-effect curve. T16Ainh-A01 did not affect the maximal contraction produced by 60 mM KCl and the relaxant effect of 10 μM T16Ainh-A01 was not altered by incubation of mouse thoracic aorta in a cocktail of potassium (K+) channel blockers. T16Ainh-A01 (10 μM) also relaxed human visceral adipose arteries by 88 ± 3%. Conclusions and Implications T16Ainh-A01 blocks calcium-activated chloride channels in vascular smooth muscle cells and relaxes murine and human blood vessels. British Journal of Pharmacology",

keywords = "Ano1, anoctamins, anti-hypertensive, calcium-activated chloride channels, patch clamp, TMEM16A, vascular smooth muscle",

author = "Davis, {Alison J.} and Jian Shi and Pritchard, {Harry A. T.} and Chadha, {Preet S.} and Normand Leblanc and Georgios Vasilikostas and Zhen Yao and Verkman, {A. S.} and Albert, {Anthony P.} and Greenwood, {Iain A.}",

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AU - Pritchard, Harry A. T.

AU - Chadha, Preet S.

AU - Leblanc, Normand

AU - Vasilikostas, Georgios

AU - Yao, Zhen

AU - Verkman, A. S.

AU - Albert, Anthony P.

AU - Greenwood, Iain A.

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