Controlling TCR and CAR activation by targeting LCK recruitment with a first-in-class small-molecule inhibitor

Autores
Woessner, Nadine M.; Chinestrad, Patricio Manuel; Rückert, Tamina; Weiß, Lena; Zintchenko, Marina; Schaffer, Anna Maria; Hartmann, Sara; Kiani, Azin; Köhn, Maja; Cabrera, Maia; Hartl, Frederike A.; Schamel, Wolfgang W.; Natalie Köhler; Lorenzano Menna, Pablo; Minguet, Susana
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
T-cell activation is driven by the recruitment of lymphocyte-specific protein tyrosine kinase (LCK) to the T-cell receptor (TCR), a critical step in initiating immune responses. Existing LCK inhibitors lack specificity because they target the conserved kinase domain shared by Src family kinases, resulting in off-target effects. Here, we introduce a novel strategy to selectively modulate T-cell activation by disrupting the interaction between the SH3 domain of LCK and the receptor kinase (RK) motif of CD3ε. Using computational modeling and high-throughput virtual screening, we identified candidate compounds targeting the SH3(LCK) domain. Functional validation revealed that one compound, C10, selectively disrupted the SH3-RK interaction, leading to reduced TCR-driven activation and proliferation, while sparing activation via alternative receptors and B-cell responses. Moreover, C10 modulated the activity of CD3ε-containing CAR and TRuC T cells, attenuating cytokine production and promoting a central-memory-like phenotype associated with enhanced persistence. These findings establish targeted disruption of LCK recruitment as a viable strategy for fine-tuning T-cell responses and propose SH3(LCK) as a druggable domain with therapeutic potential for autoimmune diseases, graft-versus-host disease, and optimizing CAR T-cell therapies.
Fil: Woessner, Nadine M.. Albert Ludwigs University of Freiburg; Alemania
Fil: Chinestrad, Patricio Manuel. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnologia. Laboratorio de Farmacologia Molecular.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Rückert, Tamina. Albert Ludwigs University of Freiburg; Alemania
Fil: Weiß, Lena. Albert Ludwigs University of Freiburg; Alemania
Fil: Zintchenko, Marina. Albert Ludwigs University of Freiburg; Alemania
Fil: Schaffer, Anna Maria. Albert Ludwigs University of Freiburg; Alemania
Fil: Hartmann, Sara. Albert Ludwigs University of Freiburg; Alemania
Fil: Kiani, Azin. Albert Ludwigs University of Freiburg; Alemania
Fil: Köhn, Maja. Albert Ludwigs University of Freiburg; Alemania
Fil: Cabrera, Maia. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnologia. Laboratorio de Farmacologia Molecular.; Argentina
Fil: Hartl, Frederike A.. Albert Ludwigs University of Freiburg; Alemania
Fil: Schamel, Wolfgang W.. Albert Ludwigs University of Freiburg; Alemania
Fil: Natalie Köhler. Albert Ludwigs University of Freiburg; Alemania
Fil: Lorenzano Menna, Pablo. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnologia. Laboratorio de Farmacologia Molecular.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Minguet, Susana. Albert Ludwigs University of Freiburg; Alemania
Materia
TCR Activation
Small molecule
Drug Design
LCK
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:11336/287784
Acceder
id CONICETDig_cbd402c7acc2f300810b406edef39a24
oai_identifier_str oai:ri.conicet.gov.ar:11336/287784
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Controlling TCR and CAR activation by targeting LCK recruitment with a first-in-class small-molecule inhibitorWoessner, Nadine M.Chinestrad, Patricio ManuelRückert, TaminaWeiß, LenaZintchenko, MarinaSchaffer, Anna MariaHartmann, SaraKiani, AzinKöhn, MajaCabrera, MaiaHartl, Frederike A.Schamel, Wolfgang W.Natalie KöhlerLorenzano Menna, PabloMinguet, SusanaTCR ActivationSmall moleculeDrug DesignLCKhttps://purl.org/becyt/ford/3.4https://purl.org/becyt/ford/3https://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3https://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3T-cell activation is driven by the recruitment of lymphocyte-specific protein tyrosine kinase (LCK) to the T-cell receptor (TCR), a critical step in initiating immune responses. Existing LCK inhibitors lack specificity because they target the conserved kinase domain shared by Src family kinases, resulting in off-target effects. Here, we introduce a novel strategy to selectively modulate T-cell activation by disrupting the interaction between the SH3 domain of LCK and the receptor kinase (RK) motif of CD3ε. Using computational modeling and high-throughput virtual screening, we identified candidate compounds targeting the SH3(LCK) domain. Functional validation revealed that one compound, C10, selectively disrupted the SH3-RK interaction, leading to reduced TCR-driven activation and proliferation, while sparing activation via alternative receptors and B-cell responses. Moreover, C10 modulated the activity of CD3ε-containing CAR and TRuC T cells, attenuating cytokine production and promoting a central-memory-like phenotype associated with enhanced persistence. These findings establish targeted disruption of LCK recruitment as a viable strategy for fine-tuning T-cell responses and propose SH3(LCK) as a druggable domain with therapeutic potential for autoimmune diseases, graft-versus-host disease, and optimizing CAR T-cell therapies.Fil: Woessner, Nadine M.. Albert Ludwigs University of Freiburg; AlemaniaFil: Chinestrad, Patricio Manuel. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnologia. Laboratorio de Farmacologia Molecular.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rückert, Tamina. Albert Ludwigs University of Freiburg; AlemaniaFil: Weiß, Lena. Albert Ludwigs University of Freiburg; AlemaniaFil: Zintchenko, Marina. Albert Ludwigs University of Freiburg; AlemaniaFil: Schaffer, Anna Maria. Albert Ludwigs University of Freiburg; AlemaniaFil: Hartmann, Sara. Albert Ludwigs University of Freiburg; AlemaniaFil: Kiani, Azin. Albert Ludwigs University of Freiburg; AlemaniaFil: Köhn, Maja. Albert Ludwigs University of Freiburg; AlemaniaFil: Cabrera, Maia. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnologia. Laboratorio de Farmacologia Molecular.; ArgentinaFil: Hartl, Frederike A.. Albert Ludwigs University of Freiburg; AlemaniaFil: Schamel, Wolfgang W.. Albert Ludwigs University of Freiburg; AlemaniaFil: Natalie Köhler. Albert Ludwigs University of Freiburg; AlemaniaFil: Lorenzano Menna, Pablo. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnologia. Laboratorio de Farmacologia Molecular.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Minguet, Susana. Albert Ludwigs University of Freiburg; AlemaniaCold Spring Harbor Laboratory Press2025-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/287784Woessner, Nadine M.; Chinestrad, Patricio Manuel; Rückert, Tamina; Weiß, Lena; Zintchenko, Marina; et al.; Controlling TCR and CAR activation by targeting LCK recruitment with a first-in-class small-molecule inhibitor; Cold Spring Harbor Laboratory Press; BioRxiv; 12-2025; 1-152692-8205CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://doi.org/10.64898/2025.12.10.693420info:eu-repo/semantics/altIdentifier/url/https://www.biorxiv.org/content/10.64898/2025.12.10.693420v1info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-06-17T09:40:22Zoai:ri.conicet.gov.ar:11336/287784instacron: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:34982026-06-17 09:40:22.38CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Controlling TCR and CAR activation by targeting LCK recruitment with a first-in-class small-molecule inhibitor
title Controlling TCR and CAR activation by targeting LCK recruitment with a first-in-class small-molecule inhibitor
spellingShingle Controlling TCR and CAR activation by targeting LCK recruitment with a first-in-class small-molecule inhibitor
Woessner, Nadine M.
TCR Activation
Small molecule
Drug Design
LCK
title_short Controlling TCR and CAR activation by targeting LCK recruitment with a first-in-class small-molecule inhibitor
title_full Controlling TCR and CAR activation by targeting LCK recruitment with a first-in-class small-molecule inhibitor
title_fullStr Controlling TCR and CAR activation by targeting LCK recruitment with a first-in-class small-molecule inhibitor
title_full_unstemmed Controlling TCR and CAR activation by targeting LCK recruitment with a first-in-class small-molecule inhibitor
title_sort Controlling TCR and CAR activation by targeting LCK recruitment with a first-in-class small-molecule inhibitor
dc.creator.none.fl_str_mv Woessner, Nadine M.
Chinestrad, Patricio Manuel
Rückert, Tamina
Weiß, Lena
Zintchenko, Marina
Schaffer, Anna Maria
Hartmann, Sara
Kiani, Azin
Köhn, Maja
Cabrera, Maia
Hartl, Frederike A.
Schamel, Wolfgang W.
Natalie Köhler
Lorenzano Menna, Pablo
Minguet, Susana
author Woessner, Nadine M.
author_facet Woessner, Nadine M.
Chinestrad, Patricio Manuel
Rückert, Tamina
Weiß, Lena
Zintchenko, Marina
Schaffer, Anna Maria
Hartmann, Sara
Kiani, Azin
Köhn, Maja
Cabrera, Maia
Hartl, Frederike A.
Schamel, Wolfgang W.
Natalie Köhler
Lorenzano Menna, Pablo
Minguet, Susana
author_role author
author2 Chinestrad, Patricio Manuel
Rückert, Tamina
Weiß, Lena
Zintchenko, Marina
Schaffer, Anna Maria
Hartmann, Sara
Kiani, Azin
Köhn, Maja
Cabrera, Maia
Hartl, Frederike A.
Schamel, Wolfgang W.
Natalie Köhler
Lorenzano Menna, Pablo
Minguet, Susana
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv TCR Activation
Small molecule
Drug Design
LCK
topic TCR Activation
Small molecule
Drug Design
LCK
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.4
https://purl.org/becyt/ford/3
https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv T-cell activation is driven by the recruitment of lymphocyte-specific protein tyrosine kinase (LCK) to the T-cell receptor (TCR), a critical step in initiating immune responses. Existing LCK inhibitors lack specificity because they target the conserved kinase domain shared by Src family kinases, resulting in off-target effects. Here, we introduce a novel strategy to selectively modulate T-cell activation by disrupting the interaction between the SH3 domain of LCK and the receptor kinase (RK) motif of CD3ε. Using computational modeling and high-throughput virtual screening, we identified candidate compounds targeting the SH3(LCK) domain. Functional validation revealed that one compound, C10, selectively disrupted the SH3-RK interaction, leading to reduced TCR-driven activation and proliferation, while sparing activation via alternative receptors and B-cell responses. Moreover, C10 modulated the activity of CD3ε-containing CAR and TRuC T cells, attenuating cytokine production and promoting a central-memory-like phenotype associated with enhanced persistence. These findings establish targeted disruption of LCK recruitment as a viable strategy for fine-tuning T-cell responses and propose SH3(LCK) as a druggable domain with therapeutic potential for autoimmune diseases, graft-versus-host disease, and optimizing CAR T-cell therapies.
Fil: Woessner, Nadine M.. Albert Ludwigs University of Freiburg; Alemania
Fil: Chinestrad, Patricio Manuel. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnologia. Laboratorio de Farmacologia Molecular.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Rückert, Tamina. Albert Ludwigs University of Freiburg; Alemania
Fil: Weiß, Lena. Albert Ludwigs University of Freiburg; Alemania
Fil: Zintchenko, Marina. Albert Ludwigs University of Freiburg; Alemania
Fil: Schaffer, Anna Maria. Albert Ludwigs University of Freiburg; Alemania
Fil: Hartmann, Sara. Albert Ludwigs University of Freiburg; Alemania
Fil: Kiani, Azin. Albert Ludwigs University of Freiburg; Alemania
Fil: Köhn, Maja. Albert Ludwigs University of Freiburg; Alemania
Fil: Cabrera, Maia. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnologia. Laboratorio de Farmacologia Molecular.; Argentina
Fil: Hartl, Frederike A.. Albert Ludwigs University of Freiburg; Alemania
Fil: Schamel, Wolfgang W.. Albert Ludwigs University of Freiburg; Alemania
Fil: Natalie Köhler. Albert Ludwigs University of Freiburg; Alemania
Fil: Lorenzano Menna, Pablo. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnologia. Laboratorio de Farmacologia Molecular.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Minguet, Susana. Albert Ludwigs University of Freiburg; Alemania
description T-cell activation is driven by the recruitment of lymphocyte-specific protein tyrosine kinase (LCK) to the T-cell receptor (TCR), a critical step in initiating immune responses. Existing LCK inhibitors lack specificity because they target the conserved kinase domain shared by Src family kinases, resulting in off-target effects. Here, we introduce a novel strategy to selectively modulate T-cell activation by disrupting the interaction between the SH3 domain of LCK and the receptor kinase (RK) motif of CD3ε. Using computational modeling and high-throughput virtual screening, we identified candidate compounds targeting the SH3(LCK) domain. Functional validation revealed that one compound, C10, selectively disrupted the SH3-RK interaction, leading to reduced TCR-driven activation and proliferation, while sparing activation via alternative receptors and B-cell responses. Moreover, C10 modulated the activity of CD3ε-containing CAR and TRuC T cells, attenuating cytokine production and promoting a central-memory-like phenotype associated with enhanced persistence. These findings establish targeted disruption of LCK recruitment as a viable strategy for fine-tuning T-cell responses and propose SH3(LCK) as a druggable domain with therapeutic potential for autoimmune diseases, graft-versus-host disease, and optimizing CAR T-cell therapies.
publishDate 2025
dc.date.none.fl_str_mv 2025-12
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/11336/287784
Woessner, Nadine M.; Chinestrad, Patricio Manuel; Rückert, Tamina; Weiß, Lena; Zintchenko, Marina; et al.; Controlling TCR and CAR activation by targeting LCK recruitment with a first-in-class small-molecule inhibitor; Cold Spring Harbor Laboratory Press; BioRxiv; 12-2025; 1-15
2692-8205
CONICET Digital
CONICET
url http://hdl.handle.net/11336/287784
identifier_str_mv Woessner, Nadine M.; Chinestrad, Patricio Manuel; Rückert, Tamina; Weiß, Lena; Zintchenko, Marina; et al.; Controlling TCR and CAR activation by targeting LCK recruitment with a first-in-class small-molecule inhibitor; Cold Spring Harbor Laboratory Press; BioRxiv; 12-2025; 1-15
2692-8205
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://doi.org/10.64898/2025.12.10.693420
info:eu-repo/semantics/altIdentifier/url/https://www.biorxiv.org/content/10.64898/2025.12.10.693420v1
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
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 13.040872

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