Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cells

Autores
Sánchez, Diana Soledad; Fisher Sigel, L. K.; Balestracci, Alejandro; Ibarra, Cristina Adriana; Amaral, María Marta; Silberstein, Claudia Marcela
Año de publicación
2020
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Shigatoxin-producing Escherichia coli isresponsible for Hemolytic Uremic Syndrome (HUS), a cause of renal failure inchildren. We have previously shown that C-9 and Eliglustat (EG), inhibitors ofglucosylceramide  synthase andglobotriaosylceramide (Gb3), prevent the cytotoxic effects of Shiga toxin type2 (Stx2), in human cortical renal tubular epithelial cells (HRTEC) primarycultures and HK2 cell line. The aim of this work was to evaluate the efficacyof EG, elucidating EG treatments necessary to achieve total protection against Stx2in HRTEC and HK2. Cells were incubated with Stx2 (1 ng/ml, 24 and 72 h) andpre-incubated with or without EG (1-500 nM, 6 and 24 h), followed byco-incubation with same dilutions of EG and Stx2 (24 and 72 h). Total number ofcells stained with Hoechst was counted in microphotographs and compared withcell viability measured by neutral red uptake. Early and late apoptosis andnecrosis was evaluated by annexin V/propidium iodide staining. Tubulogenesis wasevaluated in HRTEC grown on matrigel. Treatment of cells with Stx2significantly decreased cell confluence and viability and the number of cellsattached (p<0.001). In HRTEC, Stx2 increased early and late apoptosis, andnecrosis compared to non-treated cells (p<0.01). Furthermore, Stx2 inhibitedcell aggregation and tubulogenesis on matrigel. HRTEC preincubated with EG (50nM, 24 h or 500 nM, 6 h) totally prevented Stx2 effects on HRTEC measured as cellcount, viability, apoptosis, necrosis and tubulogenesis (p<0.05). Preincubationof HK2 cells with EG (1 nM, 24 h or 10 nM, 6 h) totally prevented Stx2 effectson cell viability and confluence. EG alone did not produce cytotoxic effectsper se. In conclusion, EG protects human renal tubular epithelium against Stx2cytotoxicity being HRTEC more sensitive than HK2. Treatment with EG could be anovel substrate inhibition therapy to neutralize Stx2 action and prevent renal damagein patients with HUS.
Fil: Sánchez, Diana Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Fisher Sigel, L. K.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Balestracci, Alejandro. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños Pedro Elizalde (ex Casa Cuna); Argentina
Fil: Ibarra, Cristina Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Amaral, María Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Silberstein, Claudia Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
LXV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXVIII Reunión Anual de la Sociedad Argentina de Inmunología y Reunión Anual de la Asociación Argentina de Fisiología
Argentina
Sociedad argentina de Investigación Clínica
Sociedad Argentina de Inmunología
Asociación Argentina de Fisiología
Materia
Eliglustat
Shiga toxin
primary culture
renal cells
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/193278
Acceder
id CONICETDig_9045533072c32344f7e45b58b5697117
oai_identifier_str oai:ri.conicet.gov.ar:11336/193278
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cellsSánchez, Diana SoledadFisher Sigel, L. K.Balestracci, AlejandroIbarra, Cristina AdrianaAmaral, María MartaSilberstein, Claudia MarcelaEliglustatShiga toxinprimary culturerenal cellshttps://purl.org/becyt/ford/3.3https://purl.org/becyt/ford/3Shigatoxin-producing Escherichia coli isresponsible for Hemolytic Uremic Syndrome (HUS), a cause of renal failure inchildren. We have previously shown that C-9 and Eliglustat (EG), inhibitors ofglucosylceramide  synthase andglobotriaosylceramide (Gb3), prevent the cytotoxic effects of Shiga toxin type2 (Stx2), in human cortical renal tubular epithelial cells (HRTEC) primarycultures and HK2 cell line. The aim of this work was to evaluate the efficacyof EG, elucidating EG treatments necessary to achieve total protection against Stx2in HRTEC and HK2. Cells were incubated with Stx2 (1 ng/ml, 24 and 72 h) andpre-incubated with or without EG (1-500 nM, 6 and 24 h), followed byco-incubation with same dilutions of EG and Stx2 (24 and 72 h). Total number ofcells stained with Hoechst was counted in microphotographs and compared withcell viability measured by neutral red uptake. Early and late apoptosis andnecrosis was evaluated by annexin V/propidium iodide staining. Tubulogenesis wasevaluated in HRTEC grown on matrigel. Treatment of cells with Stx2significantly decreased cell confluence and viability and the number of cellsattached (p<0.001). In HRTEC, Stx2 increased early and late apoptosis, andnecrosis compared to non-treated cells (p<0.01). Furthermore, Stx2 inhibitedcell aggregation and tubulogenesis on matrigel. HRTEC preincubated with EG (50nM, 24 h or 500 nM, 6 h) totally prevented Stx2 effects on HRTEC measured as cellcount, viability, apoptosis, necrosis and tubulogenesis (p<0.05). Preincubationof HK2 cells with EG (1 nM, 24 h or 10 nM, 6 h) totally prevented Stx2 effectson cell viability and confluence. EG alone did not produce cytotoxic effectsper se. In conclusion, EG protects human renal tubular epithelium against Stx2cytotoxicity being HRTEC more sensitive than HK2. Treatment with EG could be anovel substrate inhibition therapy to neutralize Stx2 action and prevent renal damagein patients with HUS.Fil: Sánchez, Diana Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Fisher Sigel, L. K.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Balestracci, Alejandro. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños Pedro Elizalde (ex Casa Cuna); ArgentinaFil: Ibarra, Cristina Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Amaral, María Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Silberstein, Claudia Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaLXV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXVIII Reunión Anual de la Sociedad Argentina de Inmunología y Reunión Anual de la Asociación Argentina de FisiologíaArgentinaSociedad argentina de Investigación ClínicaSociedad Argentina de InmunologíaAsociación Argentina de FisiologíaFundación Revista Medicina2020info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/193278Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cells; LXV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXVIII Reunión Anual de la Sociedad Argentina de Inmunología y Reunión Anual de la Asociación Argentina de Fisiología; Argentina; 2020; 100-1000025-7680CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.saic.org.ar/reunion-anualNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:46:28Zoai:ri.conicet.gov.ar:11336/193278instacron: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:34982025-09-03 09:46:28.545CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cells
title Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cells
spellingShingle Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cells
Sánchez, Diana Soledad
Eliglustat
Shiga toxin
primary culture
renal cells
title_short Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cells
title_full Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cells
title_fullStr Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cells
title_full_unstemmed Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cells
title_sort Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cells
dc.creator.none.fl_str_mv Sánchez, Diana Soledad
Fisher Sigel, L. K.
Balestracci, Alejandro
Ibarra, Cristina Adriana
Amaral, María Marta
Silberstein, Claudia Marcela
author Sánchez, Diana Soledad
author_facet Sánchez, Diana Soledad
Fisher Sigel, L. K.
Balestracci, Alejandro
Ibarra, Cristina Adriana
Amaral, María Marta
Silberstein, Claudia Marcela
author_role author
author2 Fisher Sigel, L. K.
Balestracci, Alejandro
Ibarra, Cristina Adriana
Amaral, María Marta
Silberstein, Claudia Marcela
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Eliglustat
Shiga toxin
primary culture
renal cells
topic Eliglustat
Shiga toxin
primary culture
renal cells
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.3
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Shigatoxin-producing Escherichia coli isresponsible for Hemolytic Uremic Syndrome (HUS), a cause of renal failure inchildren. We have previously shown that C-9 and Eliglustat (EG), inhibitors ofglucosylceramide  synthase andglobotriaosylceramide (Gb3), prevent the cytotoxic effects of Shiga toxin type2 (Stx2), in human cortical renal tubular epithelial cells (HRTEC) primarycultures and HK2 cell line. The aim of this work was to evaluate the efficacyof EG, elucidating EG treatments necessary to achieve total protection against Stx2in HRTEC and HK2. Cells were incubated with Stx2 (1 ng/ml, 24 and 72 h) andpre-incubated with or without EG (1-500 nM, 6 and 24 h), followed byco-incubation with same dilutions of EG and Stx2 (24 and 72 h). Total number ofcells stained with Hoechst was counted in microphotographs and compared withcell viability measured by neutral red uptake. Early and late apoptosis andnecrosis was evaluated by annexin V/propidium iodide staining. Tubulogenesis wasevaluated in HRTEC grown on matrigel. Treatment of cells with Stx2significantly decreased cell confluence and viability and the number of cellsattached (p<0.001). In HRTEC, Stx2 increased early and late apoptosis, andnecrosis compared to non-treated cells (p<0.01). Furthermore, Stx2 inhibitedcell aggregation and tubulogenesis on matrigel. HRTEC preincubated with EG (50nM, 24 h or 500 nM, 6 h) totally prevented Stx2 effects on HRTEC measured as cellcount, viability, apoptosis, necrosis and tubulogenesis (p<0.05). Preincubationof HK2 cells with EG (1 nM, 24 h or 10 nM, 6 h) totally prevented Stx2 effectson cell viability and confluence. EG alone did not produce cytotoxic effectsper se. In conclusion, EG protects human renal tubular epithelium against Stx2cytotoxicity being HRTEC more sensitive than HK2. Treatment with EG could be anovel substrate inhibition therapy to neutralize Stx2 action and prevent renal damagein patients with HUS.
Fil: Sánchez, Diana Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Fisher Sigel, L. K.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Balestracci, Alejandro. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños Pedro Elizalde (ex Casa Cuna); Argentina
Fil: Ibarra, Cristina Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Amaral, María Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Silberstein, Claudia Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
LXV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXVIII Reunión Anual de la Sociedad Argentina de Inmunología y Reunión Anual de la Asociación Argentina de Fisiología
Argentina
Sociedad argentina de Investigación Clínica
Sociedad Argentina de Inmunología
Asociación Argentina de Fisiología
description Shigatoxin-producing Escherichia coli isresponsible for Hemolytic Uremic Syndrome (HUS), a cause of renal failure inchildren. We have previously shown that C-9 and Eliglustat (EG), inhibitors ofglucosylceramide  synthase andglobotriaosylceramide (Gb3), prevent the cytotoxic effects of Shiga toxin type2 (Stx2), in human cortical renal tubular epithelial cells (HRTEC) primarycultures and HK2 cell line. The aim of this work was to evaluate the efficacyof EG, elucidating EG treatments necessary to achieve total protection against Stx2in HRTEC and HK2. Cells were incubated with Stx2 (1 ng/ml, 24 and 72 h) andpre-incubated with or without EG (1-500 nM, 6 and 24 h), followed byco-incubation with same dilutions of EG and Stx2 (24 and 72 h). Total number ofcells stained with Hoechst was counted in microphotographs and compared withcell viability measured by neutral red uptake. Early and late apoptosis andnecrosis was evaluated by annexin V/propidium iodide staining. Tubulogenesis wasevaluated in HRTEC grown on matrigel. Treatment of cells with Stx2significantly decreased cell confluence and viability and the number of cellsattached (p<0.001). In HRTEC, Stx2 increased early and late apoptosis, andnecrosis compared to non-treated cells (p<0.01). Furthermore, Stx2 inhibitedcell aggregation and tubulogenesis on matrigel. HRTEC preincubated with EG (50nM, 24 h or 500 nM, 6 h) totally prevented Stx2 effects on HRTEC measured as cellcount, viability, apoptosis, necrosis and tubulogenesis (p<0.05). Preincubationof HK2 cells with EG (1 nM, 24 h or 10 nM, 6 h) totally prevented Stx2 effectson cell viability and confluence. EG alone did not produce cytotoxic effectsper se. In conclusion, EG protects human renal tubular epithelium against Stx2cytotoxicity being HRTEC more sensitive than HK2. Treatment with EG could be anovel substrate inhibition therapy to neutralize Stx2 action and prevent renal damagein patients with HUS.
publishDate 2020
dc.date.none.fl_str_mv 2020
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/193278
Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cells; LXV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXVIII Reunión Anual de la Sociedad Argentina de Inmunología y Reunión Anual de la Asociación Argentina de Fisiología; Argentina; 2020; 100-100
0025-7680
CONICET Digital
CONICET
url http://hdl.handle.net/11336/193278
identifier_str_mv Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cells; LXV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXVIII Reunión Anual de la Sociedad Argentina de Inmunología y Reunión Anual de la Asociación Argentina de Fisiología; Argentina; 2020; 100-100
0025-7680
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
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dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
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dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Fundación Revista Medicina
publisher.none.fl_str_mv Fundación Revista Medicina
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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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