Differential Sensitivity of TRPC- and ORAI-Mediated Calcium Entries to 1‑[2-(4-Methoxyphenyl)-2-[3-(4- methoxyphenyl)propoxy]ethyl]imidazole Chloride (SKF-96365)

Autores
Susperreguy, Sebastián; Formoso, Karina; Mansilla Ricartti, Julieta; Freichel, Marc; Birnbaumer, Lutz
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Fil: Susperreguy, Sebastián. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Neurobiología Molecular; Argentina
Fil: Formoso, Karina. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Biología Celular y Molecular; Argentina
Fil: Mansilla Ricartti, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Freichel, Marc. Heidelberg University. Institute of Physiology and Pathophysiology; Alemania
Fil: Birnbaumer, Lutz. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas; Argentina
Background and Purpose: SKF-96365 is widely used as a broad-spectrum inhibitor of calcium entry. It was initially identified as a Receptor-Operated Ca2+ Entry (ROCE) blocker and was later shown to inhibit ORAI1−STIM1-mediated StoreOperated Ca2+ Entry (SOCE). However, its selectivity for TRPC versus the ORAI channels remains unclear. Experimental Approach: To examine the selectivity of SKF-96365, we evaluated its effects on SOCE and ROCE in wild-type and TRPC hepta-KO mouse embryonic fibroblasts (MEFs), as well as on TRPCmediated OAG-induced calcium entry in HEK293 cells overexpressing TRPC3, TRPC6, or TRPC7. Additional assays were conducted on HEK293 cells expressing the muscarinic M5 receptor (M5R). Half-maximal inhibitory concentration (IC50) values were determined under all conditions. Key Results: SKF-96365 suppressed thapsigargin (Tg)-induced SOCE similarly in wild-type and TRPC hepta-KO MEFs, with IC50 values around 4−5 μM. Comparable inhibition was observed for carbachol (CCh)-activated ROCE in TRPC-deficient cells. In contrast, OAG-activated Ca2+ entry by TRPC3/6/7 was only weakly inhibited, with IC50 values exceeding 100 μM. Notably, TRPC-mediated Ca2+ entry was unaffected by CRAC channel blockers or ORAI coexpression, confirming its independence of SOCE mechanisms Conclusions and Implications: Our findings demonstrate that ORAI-mediated SOCE is approximately 25-fold more sensitive to SKF-96365 than TRPC-mediated calcium entry. GSK-7975A further confirmed the ORAI selectivity by blocking SOCE without affecting TRPC channels. These results clarify the pharmacological profile of SKF96365, confirming its primary action on ORAI channels and highlight the need for concentration-aware interpretation in calcium signaling studies, particularly in the context of CRAC channel-related disorders.
Fuente
ACS Pharmacology & Translational Science, 8(12), 4237–4247
Materia
CALCIO
TRPC
BIOMEDICINA
SKF-96365
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
Repositorio Institucional (UCA)
Institución
Pontificia Universidad Católica Argentina
OAI Identificador
oai:ucacris:123456789/21905
Acceder
id RIUCA_a6709aa40c5fe496be1954513c5a1721
oai_identifier_str oai:ucacris:123456789/21905
network_acronym_str RIUCA
repository_id_str 2585
network_name_str Repositorio Institucional (UCA)
spelling Differential Sensitivity of TRPC- and ORAI-Mediated Calcium Entries to 1‑[2-(4-Methoxyphenyl)-2-[3-(4- methoxyphenyl)propoxy]ethyl]imidazole Chloride (SKF-96365)Susperreguy, SebastiánFormoso, KarinaMansilla Ricartti, JulietaFreichel, MarcBirnbaumer, LutzCALCIOTRPCBIOMEDICINASKF-96365Fil: Susperreguy, Sebastián. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Neurobiología Molecular; ArgentinaFil: Formoso, Karina. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Biología Celular y Molecular; ArgentinaFil: Mansilla Ricartti, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Freichel, Marc. Heidelberg University. Institute of Physiology and Pathophysiology; AlemaniaFil: Birnbaumer, Lutz. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas; ArgentinaBackground and Purpose: SKF-96365 is widely used as a broad-spectrum inhibitor of calcium entry. It was initially identified as a Receptor-Operated Ca2+ Entry (ROCE) blocker and was later shown to inhibit ORAI1−STIM1-mediated StoreOperated Ca2+ Entry (SOCE). However, its selectivity for TRPC versus the ORAI channels remains unclear. Experimental Approach: To examine the selectivity of SKF-96365, we evaluated its effects on SOCE and ROCE in wild-type and TRPC hepta-KO mouse embryonic fibroblasts (MEFs), as well as on TRPCmediated OAG-induced calcium entry in HEK293 cells overexpressing TRPC3, TRPC6, or TRPC7. Additional assays were conducted on HEK293 cells expressing the muscarinic M5 receptor (M5R). Half-maximal inhibitory concentration (IC50) values were determined under all conditions. Key Results: SKF-96365 suppressed thapsigargin (Tg)-induced SOCE similarly in wild-type and TRPC hepta-KO MEFs, with IC50 values around 4−5 μM. Comparable inhibition was observed for carbachol (CCh)-activated ROCE in TRPC-deficient cells. In contrast, OAG-activated Ca2+ entry by TRPC3/6/7 was only weakly inhibited, with IC50 values exceeding 100 μM. Notably, TRPC-mediated Ca2+ entry was unaffected by CRAC channel blockers or ORAI coexpression, confirming its independence of SOCE mechanisms Conclusions and Implications: Our findings demonstrate that ORAI-mediated SOCE is approximately 25-fold more sensitive to SKF-96365 than TRPC-mediated calcium entry. GSK-7975A further confirmed the ORAI selectivity by blocking SOCE without affecting TRPC channels. These results clarify the pharmacological profile of SKF96365, confirming its primary action on ORAI channels and highlight the need for concentration-aware interpretation in calcium signaling studies, particularly in the context of CRAC channel-related disorders.American Chemical Society2025info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttps://repositorio.uca.edu.ar/handle/123456789/21905https://doi.org/10.1021/acsptsci.5c00172ACS Pharmacology & Translational Science, 8(12), 4237–4247reponame:Repositorio Institucional (UCA)instname:Pontificia Universidad Católica Argentinaenginfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/4.0/2026-10-01T12:03:35Zoai:ucacris:123456789/21905instacron:UCAInstitucionalhttps://repositorio.uca.edu.ar/Universidad privadaNo correspondehttps://repositorio.uca.edu.ar/oaiclaudia_fernandez@uca.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:25852026-10-01 12:03:35.947Repositorio Institucional (UCA) - Pontificia Universidad Católica Argentinafalse
dc.title.none.fl_str_mv Differential Sensitivity of TRPC- and ORAI-Mediated Calcium Entries to 1‑[2-(4-Methoxyphenyl)-2-[3-(4- methoxyphenyl)propoxy]ethyl]imidazole Chloride (SKF-96365)
title Differential Sensitivity of TRPC- and ORAI-Mediated Calcium Entries to 1‑[2-(4-Methoxyphenyl)-2-[3-(4- methoxyphenyl)propoxy]ethyl]imidazole Chloride (SKF-96365)
spellingShingle Differential Sensitivity of TRPC- and ORAI-Mediated Calcium Entries to 1‑[2-(4-Methoxyphenyl)-2-[3-(4- methoxyphenyl)propoxy]ethyl]imidazole Chloride (SKF-96365)
Susperreguy, Sebastián
CALCIO
TRPC
BIOMEDICINA
SKF-96365
title_short Differential Sensitivity of TRPC- and ORAI-Mediated Calcium Entries to 1‑[2-(4-Methoxyphenyl)-2-[3-(4- methoxyphenyl)propoxy]ethyl]imidazole Chloride (SKF-96365)
title_full Differential Sensitivity of TRPC- and ORAI-Mediated Calcium Entries to 1‑[2-(4-Methoxyphenyl)-2-[3-(4- methoxyphenyl)propoxy]ethyl]imidazole Chloride (SKF-96365)
title_fullStr Differential Sensitivity of TRPC- and ORAI-Mediated Calcium Entries to 1‑[2-(4-Methoxyphenyl)-2-[3-(4- methoxyphenyl)propoxy]ethyl]imidazole Chloride (SKF-96365)
title_full_unstemmed Differential Sensitivity of TRPC- and ORAI-Mediated Calcium Entries to 1‑[2-(4-Methoxyphenyl)-2-[3-(4- methoxyphenyl)propoxy]ethyl]imidazole Chloride (SKF-96365)
title_sort Differential Sensitivity of TRPC- and ORAI-Mediated Calcium Entries to 1‑[2-(4-Methoxyphenyl)-2-[3-(4- methoxyphenyl)propoxy]ethyl]imidazole Chloride (SKF-96365)
dc.creator.none.fl_str_mv Susperreguy, Sebastián
Formoso, Karina
Mansilla Ricartti, Julieta
Freichel, Marc
Birnbaumer, Lutz
author Susperreguy, Sebastián
author_facet Susperreguy, Sebastián
Formoso, Karina
Mansilla Ricartti, Julieta
Freichel, Marc
Birnbaumer, Lutz
author_role author
author2 Formoso, Karina
Mansilla Ricartti, Julieta
Freichel, Marc
Birnbaumer, Lutz
author2_role author
author
author
author
dc.subject.none.fl_str_mv CALCIO
TRPC
BIOMEDICINA
SKF-96365
topic CALCIO
TRPC
BIOMEDICINA
SKF-96365
dc.description.none.fl_txt_mv Fil: Susperreguy, Sebastián. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Neurobiología Molecular; Argentina
Fil: Formoso, Karina. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Biología Celular y Molecular; Argentina
Fil: Mansilla Ricartti, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Freichel, Marc. Heidelberg University. Institute of Physiology and Pathophysiology; Alemania
Fil: Birnbaumer, Lutz. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas; Argentina
Background and Purpose: SKF-96365 is widely used as a broad-spectrum inhibitor of calcium entry. It was initially identified as a Receptor-Operated Ca2+ Entry (ROCE) blocker and was later shown to inhibit ORAI1−STIM1-mediated StoreOperated Ca2+ Entry (SOCE). However, its selectivity for TRPC versus the ORAI channels remains unclear. Experimental Approach: To examine the selectivity of SKF-96365, we evaluated its effects on SOCE and ROCE in wild-type and TRPC hepta-KO mouse embryonic fibroblasts (MEFs), as well as on TRPCmediated OAG-induced calcium entry in HEK293 cells overexpressing TRPC3, TRPC6, or TRPC7. Additional assays were conducted on HEK293 cells expressing the muscarinic M5 receptor (M5R). Half-maximal inhibitory concentration (IC50) values were determined under all conditions. Key Results: SKF-96365 suppressed thapsigargin (Tg)-induced SOCE similarly in wild-type and TRPC hepta-KO MEFs, with IC50 values around 4−5 μM. Comparable inhibition was observed for carbachol (CCh)-activated ROCE in TRPC-deficient cells. In contrast, OAG-activated Ca2+ entry by TRPC3/6/7 was only weakly inhibited, with IC50 values exceeding 100 μM. Notably, TRPC-mediated Ca2+ entry was unaffected by CRAC channel blockers or ORAI coexpression, confirming its independence of SOCE mechanisms Conclusions and Implications: Our findings demonstrate that ORAI-mediated SOCE is approximately 25-fold more sensitive to SKF-96365 than TRPC-mediated calcium entry. GSK-7975A further confirmed the ORAI selectivity by blocking SOCE without affecting TRPC channels. These results clarify the pharmacological profile of SKF96365, confirming its primary action on ORAI channels and highlight the need for concentration-aware interpretation in calcium signaling studies, particularly in the context of CRAC channel-related disorders.
description Fil: Susperreguy, Sebastián. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Neurobiología Molecular; Argentina
publishDate 2025
dc.date.none.fl_str_mv 2025
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://repositorio.uca.edu.ar/handle/123456789/21905
https://doi.org/10.1021/acsptsci.5c00172
url https://repositorio.uca.edu.ar/handle/123456789/21905
https://doi.org/10.1021/acsptsci.5c00172
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/4.0/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/4.0/
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
dc.source.none.fl_str_mv ACS Pharmacology & Translational Science, 8(12), 4237–4247
reponame:Repositorio Institucional (UCA)
instname:Pontificia Universidad Católica Argentina
reponame_str Repositorio Institucional (UCA)
collection Repositorio Institucional (UCA)
instname_str Pontificia Universidad Católica Argentina
repository.name.fl_str_mv Repositorio Institucional (UCA) - Pontificia Universidad Católica Argentina
repository.mail.fl_str_mv claudia_fernandez@uca.edu.ar
_version_ 1877862978986967040
score 12.854657

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