Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes

Autores
Scaife, Matthew; Pacienza, Natalia Alejandra; Au, B. C. Y.; Wang, J. C. M.; Devine, S.; Scheid, E.; Lee, C. J.; Lopez Perez, O.; Neschadim, A.; Fowler, D. H.; Foley, R.; Medin, J. A.
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Cell-fate control gene therapy (CFCGT)-based strategies can augment existing gene therapy and cell transplantation approaches by providing a safety element in the event of deleterious outcomes. Previously, we described a novel enzyme/prodrug combination for CFCGT. Here, we present results employing novel lentiviral constructs harboring sequences for truncated surface molecules (CD19 or low-affinity nerve growth factor receptor) directly fused to that CFCGT cDNA (TmpkF105Y). This confers an enforced one-to-one correlation between cell marking and eradication functions. In-vitro analysis demonstrated the full functionality of the fusion product. Next, low-dose 3'-azido-3'-deoxythymidine (AZT) administration to non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice injected with transduced clonal K562 cells suppressed tumor growth; furthermore, one integrated vector on average was sufficient to mediate cytotoxicity. Further, in a murine xenogeneic leukemia-lymphoma model we also demonstrated in-vivo control over transduced Raji cells. Finally, in a proof-of-principle study to examine the utility of this cassette in combination with a therapeutic cDNA, we integrated this novel CFCGT fusion construct into a lentivector designed for treatment of Fabry disease. Transduction with this vector restored enzyme activity in Fabry cells and retained AZT sensitivity. In addition, human Fabry patient CD34(+) cells showed high transduction efficiencies and retained normal colony-generating capacity when compared with the non-transduced controls. These collective results demonstrated that this novel and broadly applicable fusion system may enhance general safety in gene- and cell-based therapies.
Fil: Scaife, Matthew. University of Toronto; Canadá
Fil: Pacienza, Natalia Alejandra. University Health Network. Ontario Cancer Institute; Canadá. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Au, B. C. Y.. University Health Network. Ontario Cancer Institute; Canadá
Fil: Wang, J. C. M.. University Health Network. Ontario Cancer Institute; Canadá
Fil: Devine, S.. University of Toronto; Canadá
Fil: Scheid, E.. Mc Master University; Canadá
Fil: Lee, C. J.. University Health Network. Ontario Cancer Institute; Canadá
Fil: Lopez Perez, O.. University Health Network. Ontario Cancer Institute; Canadá
Fil: Neschadim, A.. University of Toronto; Canadá
Fil: Fowler, D. H.. National Institutes of Health; Estados Unidos
Fil: Foley, R.. Mc Master University; Canadá
Fil: Medin, J. A.. University of Toronto; Canadá. University Health Network. Ontario Cancer Institute; Canadá
Materia
Tmpk
Cell-Fate Control
Lentivirus
Gene Therapy
Azt
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:11336/22544
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/22544
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettesScaife, MatthewPacienza, Natalia AlejandraAu, B. C. Y.Wang, J. C. M.Devine, S.Scheid, E.Lee, C. J.Lopez Perez, O.Neschadim, A.Fowler, D. H.Foley, R.Medin, J. A.TmpkCell-Fate ControlLentivirusGene TherapyAzthttps://purl.org/becyt/ford/3.4https://purl.org/becyt/ford/3https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Cell-fate control gene therapy (CFCGT)-based strategies can augment existing gene therapy and cell transplantation approaches by providing a safety element in the event of deleterious outcomes. Previously, we described a novel enzyme/prodrug combination for CFCGT. Here, we present results employing novel lentiviral constructs harboring sequences for truncated surface molecules (CD19 or low-affinity nerve growth factor receptor) directly fused to that CFCGT cDNA (TmpkF105Y). This confers an enforced one-to-one correlation between cell marking and eradication functions. In-vitro analysis demonstrated the full functionality of the fusion product. Next, low-dose 3'-azido-3'-deoxythymidine (AZT) administration to non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice injected with transduced clonal K562 cells suppressed tumor growth; furthermore, one integrated vector on average was sufficient to mediate cytotoxicity. Further, in a murine xenogeneic leukemia-lymphoma model we also demonstrated in-vivo control over transduced Raji cells. Finally, in a proof-of-principle study to examine the utility of this cassette in combination with a therapeutic cDNA, we integrated this novel CFCGT fusion construct into a lentivector designed for treatment of Fabry disease. Transduction with this vector restored enzyme activity in Fabry cells and retained AZT sensitivity. In addition, human Fabry patient CD34(+) cells showed high transduction efficiencies and retained normal colony-generating capacity when compared with the non-transduced controls. These collective results demonstrated that this novel and broadly applicable fusion system may enhance general safety in gene- and cell-based therapies.Fil: Scaife, Matthew. University of Toronto; CanadáFil: Pacienza, Natalia Alejandra. University Health Network. Ontario Cancer Institute; Canadá. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Au, B. C. Y.. University Health Network. Ontario Cancer Institute; CanadáFil: Wang, J. C. M.. University Health Network. Ontario Cancer Institute; CanadáFil: Devine, S.. University of Toronto; CanadáFil: Scheid, E.. Mc Master University; CanadáFil: Lee, C. J.. University Health Network. Ontario Cancer Institute; CanadáFil: Lopez Perez, O.. University Health Network. Ontario Cancer Institute; CanadáFil: Neschadim, A.. University of Toronto; CanadáFil: Fowler, D. H.. National Institutes of Health; Estados UnidosFil: Foley, R.. Mc Master University; CanadáFil: Medin, J. A.. University of Toronto; Canadá. University Health Network. Ontario Cancer Institute; CanadáNature Publishing Group2013-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/22544Scaife, Matthew; Pacienza, Natalia Alejandra; Au, B. C. Y.; Wang, J. C. M.; Devine, S.; et al.; Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes; Nature Publishing Group; Gene Therapy; 20; 1-2013; 24-340969-71281476-5462CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.nature.com/gt/journal/v20/n1/full/gt2011210a.html?foxtrotcallback=trueinfo:eu-repo/semantics/altIdentifier/doi/10.1038/gt.2011.210info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:55:18Zoai:ri.conicet.gov.ar:11336/22544instacron: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:34982025-10-15 14:55:18.856CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes
title Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes
spellingShingle Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes
Scaife, Matthew
Tmpk
Cell-Fate Control
Lentivirus
Gene Therapy
Azt
title_short Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes
title_full Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes
title_fullStr Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes
title_full_unstemmed Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes
title_sort Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes
dc.creator.none.fl_str_mv Scaife, Matthew
Pacienza, Natalia Alejandra
Au, B. C. Y.
Wang, J. C. M.
Devine, S.
Scheid, E.
Lee, C. J.
Lopez Perez, O.
Neschadim, A.
Fowler, D. H.
Foley, R.
Medin, J. A.
author Scaife, Matthew
author_facet Scaife, Matthew
Pacienza, Natalia Alejandra
Au, B. C. Y.
Wang, J. C. M.
Devine, S.
Scheid, E.
Lee, C. J.
Lopez Perez, O.
Neschadim, A.
Fowler, D. H.
Foley, R.
Medin, J. A.
author_role author
author2 Pacienza, Natalia Alejandra
Au, B. C. Y.
Wang, J. C. M.
Devine, S.
Scheid, E.
Lee, C. J.
Lopez Perez, O.
Neschadim, A.
Fowler, D. H.
Foley, R.
Medin, J. A.
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Tmpk
Cell-Fate Control
Lentivirus
Gene Therapy
Azt
topic Tmpk
Cell-Fate Control
Lentivirus
Gene Therapy
Azt
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.4
https://purl.org/becyt/ford/3
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Cell-fate control gene therapy (CFCGT)-based strategies can augment existing gene therapy and cell transplantation approaches by providing a safety element in the event of deleterious outcomes. Previously, we described a novel enzyme/prodrug combination for CFCGT. Here, we present results employing novel lentiviral constructs harboring sequences for truncated surface molecules (CD19 or low-affinity nerve growth factor receptor) directly fused to that CFCGT cDNA (TmpkF105Y). This confers an enforced one-to-one correlation between cell marking and eradication functions. In-vitro analysis demonstrated the full functionality of the fusion product. Next, low-dose 3'-azido-3'-deoxythymidine (AZT) administration to non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice injected with transduced clonal K562 cells suppressed tumor growth; furthermore, one integrated vector on average was sufficient to mediate cytotoxicity. Further, in a murine xenogeneic leukemia-lymphoma model we also demonstrated in-vivo control over transduced Raji cells. Finally, in a proof-of-principle study to examine the utility of this cassette in combination with a therapeutic cDNA, we integrated this novel CFCGT fusion construct into a lentivector designed for treatment of Fabry disease. Transduction with this vector restored enzyme activity in Fabry cells and retained AZT sensitivity. In addition, human Fabry patient CD34(+) cells showed high transduction efficiencies and retained normal colony-generating capacity when compared with the non-transduced controls. These collective results demonstrated that this novel and broadly applicable fusion system may enhance general safety in gene- and cell-based therapies.
Fil: Scaife, Matthew. University of Toronto; Canadá
Fil: Pacienza, Natalia Alejandra. University Health Network. Ontario Cancer Institute; Canadá. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Au, B. C. Y.. University Health Network. Ontario Cancer Institute; Canadá
Fil: Wang, J. C. M.. University Health Network. Ontario Cancer Institute; Canadá
Fil: Devine, S.. University of Toronto; Canadá
Fil: Scheid, E.. Mc Master University; Canadá
Fil: Lee, C. J.. University Health Network. Ontario Cancer Institute; Canadá
Fil: Lopez Perez, O.. University Health Network. Ontario Cancer Institute; Canadá
Fil: Neschadim, A.. University of Toronto; Canadá
Fil: Fowler, D. H.. National Institutes of Health; Estados Unidos
Fil: Foley, R.. Mc Master University; Canadá
Fil: Medin, J. A.. University of Toronto; Canadá. University Health Network. Ontario Cancer Institute; Canadá
description Cell-fate control gene therapy (CFCGT)-based strategies can augment existing gene therapy and cell transplantation approaches by providing a safety element in the event of deleterious outcomes. Previously, we described a novel enzyme/prodrug combination for CFCGT. Here, we present results employing novel lentiviral constructs harboring sequences for truncated surface molecules (CD19 or low-affinity nerve growth factor receptor) directly fused to that CFCGT cDNA (TmpkF105Y). This confers an enforced one-to-one correlation between cell marking and eradication functions. In-vitro analysis demonstrated the full functionality of the fusion product. Next, low-dose 3'-azido-3'-deoxythymidine (AZT) administration to non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice injected with transduced clonal K562 cells suppressed tumor growth; furthermore, one integrated vector on average was sufficient to mediate cytotoxicity. Further, in a murine xenogeneic leukemia-lymphoma model we also demonstrated in-vivo control over transduced Raji cells. Finally, in a proof-of-principle study to examine the utility of this cassette in combination with a therapeutic cDNA, we integrated this novel CFCGT fusion construct into a lentivector designed for treatment of Fabry disease. Transduction with this vector restored enzyme activity in Fabry cells and retained AZT sensitivity. In addition, human Fabry patient CD34(+) cells showed high transduction efficiencies and retained normal colony-generating capacity when compared with the non-transduced controls. These collective results demonstrated that this novel and broadly applicable fusion system may enhance general safety in gene- and cell-based therapies.
publishDate 2013
dc.date.none.fl_str_mv 2013-01
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/11336/22544
Scaife, Matthew; Pacienza, Natalia Alejandra; Au, B. C. Y.; Wang, J. C. M.; Devine, S.; et al.; Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes; Nature Publishing Group; Gene Therapy; 20; 1-2013; 24-34
0969-7128
1476-5462
CONICET Digital
CONICET
url http://hdl.handle.net/11336/22544
identifier_str_mv Scaife, Matthew; Pacienza, Natalia Alejandra; Au, B. C. Y.; Wang, J. C. M.; Devine, S.; et al.; Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes; Nature Publishing Group; Gene Therapy; 20; 1-2013; 24-34
0969-7128
1476-5462
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.nature.com/gt/journal/v20/n1/full/gt2011210a.html?foxtrotcallback=true
info:eu-repo/semantics/altIdentifier/doi/10.1038/gt.2011.210
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 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 13.22299

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