R534C mutation in hERG causes a trafficking defect in iPSC-derived cardiomyocytes from patients with type 2 long QT syndrome

Autores
Mesquita, Fernanda C. P.; Arantes, Paulo C.; Kasai Brunswick, Tais H.; Araujo, Dayana S.; Gubert, Fernanda; Monnerat, Gustavo; Silva dos Santos, Danúbia; Neiman, Gabriel; Leitão, Isabela C.; Barbosa, Raiana A. Q.; Coutinho, Jorge L.; Vaz, Isadora M.; dos Santos, Marcus N.; Borgonovo, Tamara; Cruz, Fernando E. S.; Miriuka, Santiago Gabriel; Medei, Emiliano H.; Campos de Carvalho, Antonio C.; Carvalho, Adriana B.
Año de publicación
2019
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Patient-specific cardiomyocytes obtained from induced pluripotent stem cells (CM-iPSC) offer unprecedented mechanistic insights in the study of inherited cardiac diseases. The objective of this work was to study a type 2 long QT syndrome (LQTS2)-associated mutation (c.1600C > T in KCNH2, p.R534C in hERG) in CM-iPSC. Peripheral blood mononuclear cells were isolated from two patients with the R534C mutation and iPSCs were generated. In addition, the same mutation was inserted in a control iPSC line by genome editing using CRISPR/Cas9. Cells expressed pluripotency markers and showed spontaneous differentiation into the three embryonic germ layers. Electrophysiology demonstrated that action potential duration (APD) of LQTS2 CM-iPSC was significantly longer than that of the control line, as well as the triangulation of the action potentials (AP), implying a longer duration of phase 3. Treatment with the IKr inhibitor E4031 only caused APD prolongation in the control line. Patch clamp showed a reduction of IKr on LQTS2 CM-iPSC compared to control, but channel activation was not significantly affected. Immunofluorescence for hERG demonstrated perinuclear staining in LQTS2 CM-iPSC. In conclusion, CM-iPSC recapitulated the LQTS2 phenotype and our findings suggest that the R534C mutation in KCNH2 leads to a channel trafficking defect to the plasma membrane.
Fil: Mesquita, Fernanda C. P.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Arantes, Paulo C.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Kasai Brunswick, Tais H.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Araujo, Dayana S.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Gubert, Fernanda. Universidade Federal do Rio de Janeiro; Brasil
Fil: Monnerat, Gustavo. Universidade Federal do Rio de Janeiro; Brasil
Fil: Silva dos Santos, Danúbia. Universidade Federal do Rio de Janeiro; Brasil
Fil: Neiman, Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina
Fil: Leitão, Isabela C.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Barbosa, Raiana A. Q.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Coutinho, Jorge L.. National Institute Of Cardiology; Brasil
Fil: Vaz, Isadora M.. Pontificia Universidad Catolica de Parana; Brasil
Fil: dos Santos, Marcus N.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Borgonovo, Tamara. Pontificia Universidad Catolica de Parana; Brasil
Fil: Cruz, Fernando E. S.. National Institute of Cardiology; Brasil
Fil: Miriuka, Santiago Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina
Fil: Medei, Emiliano H.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Campos de Carvalho, Antonio C.. Universidade Federal do Rio de Janeiro; Brasil. National Institute of Cardiology; Brasil. National Institute for Science and Technology in Regenerative Medicine; Brasil
Fil: Carvalho, Adriana B.. Universidade Federal do Rio de Janeiro; Brasil. National Institute for Science and Technology in Regenerative Medicine; Brasil
Materia
LONG QT SYNDROME
IPS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/175631
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/175631
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling R534C mutation in hERG causes a trafficking defect in iPSC-derived cardiomyocytes from patients with type 2 long QT syndromeMesquita, Fernanda C. P.Arantes, Paulo C.Kasai Brunswick, Tais H.Araujo, Dayana S.Gubert, FernandaMonnerat, GustavoSilva dos Santos, DanúbiaNeiman, GabrielLeitão, Isabela C.Barbosa, Raiana A. Q.Coutinho, Jorge L.Vaz, Isadora M.dos Santos, Marcus N.Borgonovo, TamaraCruz, Fernando E. S.Miriuka, Santiago GabrielMedei, Emiliano H.Campos de Carvalho, Antonio C.Carvalho, Adriana B.LONG QT SYNDROMEIPShttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Patient-specific cardiomyocytes obtained from induced pluripotent stem cells (CM-iPSC) offer unprecedented mechanistic insights in the study of inherited cardiac diseases. The objective of this work was to study a type 2 long QT syndrome (LQTS2)-associated mutation (c.1600C > T in KCNH2, p.R534C in hERG) in CM-iPSC. Peripheral blood mononuclear cells were isolated from two patients with the R534C mutation and iPSCs were generated. In addition, the same mutation was inserted in a control iPSC line by genome editing using CRISPR/Cas9. Cells expressed pluripotency markers and showed spontaneous differentiation into the three embryonic germ layers. Electrophysiology demonstrated that action potential duration (APD) of LQTS2 CM-iPSC was significantly longer than that of the control line, as well as the triangulation of the action potentials (AP), implying a longer duration of phase 3. Treatment with the IKr inhibitor E4031 only caused APD prolongation in the control line. Patch clamp showed a reduction of IKr on LQTS2 CM-iPSC compared to control, but channel activation was not significantly affected. Immunofluorescence for hERG demonstrated perinuclear staining in LQTS2 CM-iPSC. In conclusion, CM-iPSC recapitulated the LQTS2 phenotype and our findings suggest that the R534C mutation in KCNH2 leads to a channel trafficking defect to the plasma membrane.Fil: Mesquita, Fernanda C. P.. Universidade Federal do Rio de Janeiro; BrasilFil: Arantes, Paulo C.. Universidade Federal do Rio de Janeiro; BrasilFil: Kasai Brunswick, Tais H.. Universidade Federal do Rio de Janeiro; BrasilFil: Araujo, Dayana S.. Universidade Federal do Rio de Janeiro; BrasilFil: Gubert, Fernanda. Universidade Federal do Rio de Janeiro; BrasilFil: Monnerat, Gustavo. Universidade Federal do Rio de Janeiro; BrasilFil: Silva dos Santos, Danúbia. Universidade Federal do Rio de Janeiro; BrasilFil: Neiman, Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; ArgentinaFil: Leitão, Isabela C.. Universidade Federal do Rio de Janeiro; BrasilFil: Barbosa, Raiana A. Q.. Universidade Federal do Rio de Janeiro; BrasilFil: Coutinho, Jorge L.. National Institute Of Cardiology; BrasilFil: Vaz, Isadora M.. Pontificia Universidad Catolica de Parana; BrasilFil: dos Santos, Marcus N.. Universidade Federal do Rio de Janeiro; BrasilFil: Borgonovo, Tamara. Pontificia Universidad Catolica de Parana; BrasilFil: Cruz, Fernando E. S.. National Institute of Cardiology; BrasilFil: Miriuka, Santiago Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; ArgentinaFil: Medei, Emiliano H.. Universidade Federal do Rio de Janeiro; BrasilFil: Campos de Carvalho, Antonio C.. Universidade Federal do Rio de Janeiro; Brasil. National Institute of Cardiology; Brasil. National Institute for Science and Technology in Regenerative Medicine; BrasilFil: Carvalho, Adriana B.. Universidade Federal do Rio de Janeiro; Brasil. National Institute for Science and Technology in Regenerative Medicine; BrasilNature Publishing Group2019-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/175631Mesquita, Fernanda C. P.; Arantes, Paulo C.; Kasai Brunswick, Tais H.; Araujo, Dayana S.; Gubert, Fernanda; et al.; R534C mutation in hERG causes a trafficking defect in iPSC-derived cardiomyocytes from patients with type 2 long QT syndrome; Nature Publishing Group; Scientific Reports; 9; 1; 12-2019; 1-92045-2322CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.nature.com/articles/s41598-019-55837-winfo:eu-repo/semantics/altIdentifier/doi/10.1038/s41598-019-55837-winfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:36:02Zoai:ri.conicet.gov.ar:11336/175631instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-22 11:36:02.425CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv R534C mutation in hERG causes a trafficking defect in iPSC-derived cardiomyocytes from patients with type 2 long QT syndrome
title R534C mutation in hERG causes a trafficking defect in iPSC-derived cardiomyocytes from patients with type 2 long QT syndrome
spellingShingle R534C mutation in hERG causes a trafficking defect in iPSC-derived cardiomyocytes from patients with type 2 long QT syndrome
Mesquita, Fernanda C. P.
LONG QT SYNDROME
IPS
title_short R534C mutation in hERG causes a trafficking defect in iPSC-derived cardiomyocytes from patients with type 2 long QT syndrome
title_full R534C mutation in hERG causes a trafficking defect in iPSC-derived cardiomyocytes from patients with type 2 long QT syndrome
title_fullStr R534C mutation in hERG causes a trafficking defect in iPSC-derived cardiomyocytes from patients with type 2 long QT syndrome
title_full_unstemmed R534C mutation in hERG causes a trafficking defect in iPSC-derived cardiomyocytes from patients with type 2 long QT syndrome
title_sort R534C mutation in hERG causes a trafficking defect in iPSC-derived cardiomyocytes from patients with type 2 long QT syndrome
dc.creator.none.fl_str_mv Mesquita, Fernanda C. P.
Arantes, Paulo C.
Kasai Brunswick, Tais H.
Araujo, Dayana S.
Gubert, Fernanda
Monnerat, Gustavo
Silva dos Santos, Danúbia
Neiman, Gabriel
Leitão, Isabela C.
Barbosa, Raiana A. Q.
Coutinho, Jorge L.
Vaz, Isadora M.
dos Santos, Marcus N.
Borgonovo, Tamara
Cruz, Fernando E. S.
Miriuka, Santiago Gabriel
Medei, Emiliano H.
Campos de Carvalho, Antonio C.
Carvalho, Adriana B.
author Mesquita, Fernanda C. P.
author_facet Mesquita, Fernanda C. P.
Arantes, Paulo C.
Kasai Brunswick, Tais H.
Araujo, Dayana S.
Gubert, Fernanda
Monnerat, Gustavo
Silva dos Santos, Danúbia
Neiman, Gabriel
Leitão, Isabela C.
Barbosa, Raiana A. Q.
Coutinho, Jorge L.
Vaz, Isadora M.
dos Santos, Marcus N.
Borgonovo, Tamara
Cruz, Fernando E. S.
Miriuka, Santiago Gabriel
Medei, Emiliano H.
Campos de Carvalho, Antonio C.
Carvalho, Adriana B.
author_role author
author2 Arantes, Paulo C.
Kasai Brunswick, Tais H.
Araujo, Dayana S.
Gubert, Fernanda
Monnerat, Gustavo
Silva dos Santos, Danúbia
Neiman, Gabriel
Leitão, Isabela C.
Barbosa, Raiana A. Q.
Coutinho, Jorge L.
Vaz, Isadora M.
dos Santos, Marcus N.
Borgonovo, Tamara
Cruz, Fernando E. S.
Miriuka, Santiago Gabriel
Medei, Emiliano H.
Campos de Carvalho, Antonio C.
Carvalho, Adriana B.
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv LONG QT SYNDROME
IPS
topic LONG QT SYNDROME
IPS
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Patient-specific cardiomyocytes obtained from induced pluripotent stem cells (CM-iPSC) offer unprecedented mechanistic insights in the study of inherited cardiac diseases. The objective of this work was to study a type 2 long QT syndrome (LQTS2)-associated mutation (c.1600C > T in KCNH2, p.R534C in hERG) in CM-iPSC. Peripheral blood mononuclear cells were isolated from two patients with the R534C mutation and iPSCs were generated. In addition, the same mutation was inserted in a control iPSC line by genome editing using CRISPR/Cas9. Cells expressed pluripotency markers and showed spontaneous differentiation into the three embryonic germ layers. Electrophysiology demonstrated that action potential duration (APD) of LQTS2 CM-iPSC was significantly longer than that of the control line, as well as the triangulation of the action potentials (AP), implying a longer duration of phase 3. Treatment with the IKr inhibitor E4031 only caused APD prolongation in the control line. Patch clamp showed a reduction of IKr on LQTS2 CM-iPSC compared to control, but channel activation was not significantly affected. Immunofluorescence for hERG demonstrated perinuclear staining in LQTS2 CM-iPSC. In conclusion, CM-iPSC recapitulated the LQTS2 phenotype and our findings suggest that the R534C mutation in KCNH2 leads to a channel trafficking defect to the plasma membrane.
Fil: Mesquita, Fernanda C. P.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Arantes, Paulo C.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Kasai Brunswick, Tais H.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Araujo, Dayana S.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Gubert, Fernanda. Universidade Federal do Rio de Janeiro; Brasil
Fil: Monnerat, Gustavo. Universidade Federal do Rio de Janeiro; Brasil
Fil: Silva dos Santos, Danúbia. Universidade Federal do Rio de Janeiro; Brasil
Fil: Neiman, Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina
Fil: Leitão, Isabela C.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Barbosa, Raiana A. Q.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Coutinho, Jorge L.. National Institute Of Cardiology; Brasil
Fil: Vaz, Isadora M.. Pontificia Universidad Catolica de Parana; Brasil
Fil: dos Santos, Marcus N.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Borgonovo, Tamara. Pontificia Universidad Catolica de Parana; Brasil
Fil: Cruz, Fernando E. S.. National Institute of Cardiology; Brasil
Fil: Miriuka, Santiago Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina
Fil: Medei, Emiliano H.. Universidade Federal do Rio de Janeiro; Brasil
Fil: Campos de Carvalho, Antonio C.. Universidade Federal do Rio de Janeiro; Brasil. National Institute of Cardiology; Brasil. National Institute for Science and Technology in Regenerative Medicine; Brasil
Fil: Carvalho, Adriana B.. Universidade Federal do Rio de Janeiro; Brasil. National Institute for Science and Technology in Regenerative Medicine; Brasil
description Patient-specific cardiomyocytes obtained from induced pluripotent stem cells (CM-iPSC) offer unprecedented mechanistic insights in the study of inherited cardiac diseases. The objective of this work was to study a type 2 long QT syndrome (LQTS2)-associated mutation (c.1600C > T in KCNH2, p.R534C in hERG) in CM-iPSC. Peripheral blood mononuclear cells were isolated from two patients with the R534C mutation and iPSCs were generated. In addition, the same mutation was inserted in a control iPSC line by genome editing using CRISPR/Cas9. Cells expressed pluripotency markers and showed spontaneous differentiation into the three embryonic germ layers. Electrophysiology demonstrated that action potential duration (APD) of LQTS2 CM-iPSC was significantly longer than that of the control line, as well as the triangulation of the action potentials (AP), implying a longer duration of phase 3. Treatment with the IKr inhibitor E4031 only caused APD prolongation in the control line. Patch clamp showed a reduction of IKr on LQTS2 CM-iPSC compared to control, but channel activation was not significantly affected. Immunofluorescence for hERG demonstrated perinuclear staining in LQTS2 CM-iPSC. In conclusion, CM-iPSC recapitulated the LQTS2 phenotype and our findings suggest that the R534C mutation in KCNH2 leads to a channel trafficking defect to the plasma membrane.
publishDate 2019
dc.date.none.fl_str_mv 2019-12
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 http://hdl.handle.net/11336/175631
Mesquita, Fernanda C. P.; Arantes, Paulo C.; Kasai Brunswick, Tais H.; Araujo, Dayana S.; Gubert, Fernanda; et al.; R534C mutation in hERG causes a trafficking defect in iPSC-derived cardiomyocytes from patients with type 2 long QT syndrome; Nature Publishing Group; Scientific Reports; 9; 1; 12-2019; 1-9
2045-2322
CONICET Digital
CONICET
url http://hdl.handle.net/11336/175631
identifier_str_mv Mesquita, Fernanda C. P.; Arantes, Paulo C.; Kasai Brunswick, Tais H.; Araujo, Dayana S.; Gubert, Fernanda; et al.; R534C mutation in hERG causes a trafficking defect in iPSC-derived cardiomyocytes from patients with type 2 long QT syndrome; Nature Publishing Group; Scientific Reports; 9; 1; 12-2019; 1-9
2045-2322
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.nature.com/articles/s41598-019-55837-w
info:eu-repo/semantics/altIdentifier/doi/10.1038/s41598-019-55837-w
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 12.982451

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