Development of phenotypic assays for identifying novel blockers of L-type calcium channels in neurons

Abstract L-type calcium channels (LTCCs) are highly expressed in the heart and brain and are critical for cardiac and neuronal functions. LTCC-blocking drugs have a long and successful record in the clinic for treating cardiovascular disorders. In contrast, establishment of their efficacy for indica...

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Autores principales:	Rebecca Hagan, Elizabeth Rex, David Woody, Monika Milewski, Thomas Glaza, Michael P. Maher, Yi Liu
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d47221df885344dc960f53333798321c
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spelling	oai:doaj.org-article:d47221df885344dc960f53333798321c2021-12-02T14:01:20ZDevelopment of phenotypic assays for identifying novel blockers of L-type calcium channels in neurons10.1038/s41598-020-80692-52045-2322https://doaj.org/article/d47221df885344dc960f53333798321c2021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-80692-5https://doaj.org/toc/2045-2322Abstract L-type calcium channels (LTCCs) are highly expressed in the heart and brain and are critical for cardiac and neuronal functions. LTCC-blocking drugs have a long and successful record in the clinic for treating cardiovascular disorders. In contrast, establishment of their efficacy for indications of the central nervous system remains challenging given the tendency of existing LTCC drugs being functionally and mechanistically more selective for peripheral tissues. LTCCs in vivo are large macromolecular complexes consisting of a pore-forming subunit and other modulatory proteins, some of which may be neuro-specific and potentially harbor mechanisms for neuronal selectivity. To exploit the possibility of identifying mechanistically novel and/or neuro-selective blockers, we developed two phenotypic assays—a calcium flux-based primary screening assay and a patch clamp secondary assay, using rat primary cortical cultures. We screened a library comprised of 1278 known bioactive agents and successfully identified a majority of the potent LTCC-blocking drugs in the library. Significantly, we identified a previously unrecognized LTCC blocker with a novel mechanism, which was corroborated by patch clamp and binding studies. As such, these phenotypic assays are robust and represent an important step towards identifying mechanistically novel and neuro-selective LTCC blockers.Rebecca HaganElizabeth RexDavid WoodyMonika MilewskiThomas GlazaMichael P. MaherYi LiuNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rebecca Hagan Elizabeth Rex David Woody Monika Milewski Thomas Glaza Michael P. Maher Yi Liu Development of phenotypic assays for identifying novel blockers of L-type calcium channels in neurons
description	Abstract L-type calcium channels (LTCCs) are highly expressed in the heart and brain and are critical for cardiac and neuronal functions. LTCC-blocking drugs have a long and successful record in the clinic for treating cardiovascular disorders. In contrast, establishment of their efficacy for indications of the central nervous system remains challenging given the tendency of existing LTCC drugs being functionally and mechanistically more selective for peripheral tissues. LTCCs in vivo are large macromolecular complexes consisting of a pore-forming subunit and other modulatory proteins, some of which may be neuro-specific and potentially harbor mechanisms for neuronal selectivity. To exploit the possibility of identifying mechanistically novel and/or neuro-selective blockers, we developed two phenotypic assays—a calcium flux-based primary screening assay and a patch clamp secondary assay, using rat primary cortical cultures. We screened a library comprised of 1278 known bioactive agents and successfully identified a majority of the potent LTCC-blocking drugs in the library. Significantly, we identified a previously unrecognized LTCC blocker with a novel mechanism, which was corroborated by patch clamp and binding studies. As such, these phenotypic assays are robust and represent an important step towards identifying mechanistically novel and neuro-selective LTCC blockers.
format	article
author	Rebecca Hagan Elizabeth Rex David Woody Monika Milewski Thomas Glaza Michael P. Maher Yi Liu
author_facet	Rebecca Hagan Elizabeth Rex David Woody Monika Milewski Thomas Glaza Michael P. Maher Yi Liu
author_sort	Rebecca Hagan
title	Development of phenotypic assays for identifying novel blockers of L-type calcium channels in neurons
title_short	Development of phenotypic assays for identifying novel blockers of L-type calcium channels in neurons
title_full	Development of phenotypic assays for identifying novel blockers of L-type calcium channels in neurons
title_fullStr	Development of phenotypic assays for identifying novel blockers of L-type calcium channels in neurons
title_full_unstemmed	Development of phenotypic assays for identifying novel blockers of L-type calcium channels in neurons
title_sort	development of phenotypic assays for identifying novel blockers of l-type calcium channels in neurons
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/d47221df885344dc960f53333798321c
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Development of phenotypic assays for identifying novel blockers of L-type calcium channels in neurons

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