Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids

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We have developed multicellular spheroids (MCS) established from LM05e and LM3 spontaneous Balb/c-murine mammary adenocarcinoma and B16 C57-murine melanoma derived cell lines as an in vitro model to study the efficacy of the herpes simplex virus thymidine kinase/ganciclovir (HSVtk/GCV) suicide syste...

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Autores principales:	Finocchiaro, L.M.E., Bumaschny, V.F., Karara, A.L., Fiszman, G.L., Casais, C.C., Glikin, G.C.
Formato:	JOUR
Materias:	Lipofection Murine adenocarcinoma Suicide gene therapy ganciclovir suicide substrate thymidine kinase animal cell animal tissue apoptosis article breast adenocarcinoma bystander effect cell growth cell proliferation cell specificity cell type controlled study drug cytotoxicity drug efficacy gene expression regulation gene transfer Herpes simplex virus in vitro study in vivo study intracellular transport melanoma cell mitochondrion monolayer culture mouse multicellular spheroid nonhuman phenotype population structure priority journal protein assembly sensitivity analysis statistical significance transgene virus resistance Adenocarcinoma Animals Antiviral Agents bcl-2-Associated X Protein beta-Galactosidase Bystander Effect Cell Division Cell Line, Tumor Drug Resistance Ganciclovir Gene Therapy Mammary Neoplasms, Animal Melanoma Mice Mice, Inbred BALB C Proto-Oncogene Proteins c-bcl-2 Simplexvirus Spheroids, Cellular Thymidine Kinase Transfection Viral Proteins Animalia Murinae
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_09291903_v11_n5_p333_Finocchiaro
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	todo:paper_09291903_v11_n5_p333_Finocchiaro2023-10-03T15:47:34Z Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids Finocchiaro, L.M.E. Bumaschny, V.F. Karara, A.L. Fiszman, G.L. Casais, C.C. Glikin, G.C. Lipofection Murine adenocarcinoma Suicide gene therapy ganciclovir suicide substrate thymidine kinase animal cell animal tissue apoptosis article breast adenocarcinoma bystander effect cell growth cell proliferation cell specificity cell type controlled study drug cytotoxicity drug efficacy gene expression regulation gene transfer Herpes simplex virus in vitro study in vivo study intracellular transport melanoma cell mitochondrion monolayer culture mouse multicellular spheroid nonhuman phenotype population structure priority journal protein assembly sensitivity analysis statistical significance transgene virus resistance Adenocarcinoma Animals Antiviral Agents bcl-2-Associated X Protein beta-Galactosidase Bystander Effect Cell Division Cell Line, Tumor Drug Resistance Ganciclovir Gene Therapy Mammary Neoplasms, Animal Melanoma Mice Mice, Inbred BALB C Proto-Oncogene Proteins c-bcl-2 Simplexvirus Spheroids, Cellular Thymidine Kinase Transfection Viral Proteins Animalia Murinae Simplexvirus We have developed multicellular spheroids (MCS) established from LM05e and LM3 spontaneous Balb/c-murine mammary adenocarcinoma and B16 C57-murine melanoma derived cell lines as an in vitro model to study the efficacy of the herpes simplex virus thymidine kinase/ganciclovir (HSVtk/GCV) suicide system. We demonstrated for the first time that HSVtk-expressing cells assembled as MCS manifested a GCV resistance phenotype compared to the same cells grown as sparse monolayers. HSVtk-expressing LM05e, LM3 and B16 spheroids were 16-, three- and nine-fold less sensitive to GCV than their respective monolayers, even though they could express transgenes 10-, eight- and five-fold more efficiently. Mixed populations of HSVtk- and their respective βgal-expressing cells displayed a cell-type specific bystander effect that was higher in monolayers than in MCS. However, HSVtk-expressing cells in two- or three-dimensional cultures were always significantly more sensitive to GCV than the βgal-expressing counterparts, supporting the feasibility of this suicide approach in vivo. We present evidence showing that HSVtk-expressing tumor cells, when transferred from monolayers to MCS, displayed: (i) lower GCV cytotoxic activity and bystander effect; (ii) higher and efficient expression of genes transferred as lipoplexes; (iii) lower cell proliferation rates; and (iv) changes in intracellular Bax/Bcl-xL rheostat of mitochondria-mediated apoptosis. Fil:Finocchiaro, L.M.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Casais, C.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Glikin, G.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_09291903_v11_n5_p333_Finocchiaro
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Lipofection Murine adenocarcinoma Suicide gene therapy ganciclovir suicide substrate thymidine kinase animal cell animal tissue apoptosis article breast adenocarcinoma bystander effect cell growth cell proliferation cell specificity cell type controlled study drug cytotoxicity drug efficacy gene expression regulation gene transfer Herpes simplex virus in vitro study in vivo study intracellular transport melanoma cell mitochondrion monolayer culture mouse multicellular spheroid nonhuman phenotype population structure priority journal protein assembly sensitivity analysis statistical significance transgene virus resistance Adenocarcinoma Animals Antiviral Agents bcl-2-Associated X Protein beta-Galactosidase Bystander Effect Cell Division Cell Line, Tumor Drug Resistance Ganciclovir Gene Therapy Mammary Neoplasms, Animal Melanoma Mice Mice, Inbred BALB C Proto-Oncogene Proteins c-bcl-2 Simplexvirus Spheroids, Cellular Thymidine Kinase Transfection Viral Proteins Animalia Murinae Simplexvirus
spellingShingle	Lipofection Murine adenocarcinoma Suicide gene therapy ganciclovir suicide substrate thymidine kinase animal cell animal tissue apoptosis article breast adenocarcinoma bystander effect cell growth cell proliferation cell specificity cell type controlled study drug cytotoxicity drug efficacy gene expression regulation gene transfer Herpes simplex virus in vitro study in vivo study intracellular transport melanoma cell mitochondrion monolayer culture mouse multicellular spheroid nonhuman phenotype population structure priority journal protein assembly sensitivity analysis statistical significance transgene virus resistance Adenocarcinoma Animals Antiviral Agents bcl-2-Associated X Protein beta-Galactosidase Bystander Effect Cell Division Cell Line, Tumor Drug Resistance Ganciclovir Gene Therapy Mammary Neoplasms, Animal Melanoma Mice Mice, Inbred BALB C Proto-Oncogene Proteins c-bcl-2 Simplexvirus Spheroids, Cellular Thymidine Kinase Transfection Viral Proteins Animalia Murinae Simplexvirus Finocchiaro, L.M.E. Bumaschny, V.F. Karara, A.L. Fiszman, G.L. Casais, C.C. Glikin, G.C. Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
topic_facet	Lipofection Murine adenocarcinoma Suicide gene therapy ganciclovir suicide substrate thymidine kinase animal cell animal tissue apoptosis article breast adenocarcinoma bystander effect cell growth cell proliferation cell specificity cell type controlled study drug cytotoxicity drug efficacy gene expression regulation gene transfer Herpes simplex virus in vitro study in vivo study intracellular transport melanoma cell mitochondrion monolayer culture mouse multicellular spheroid nonhuman phenotype population structure priority journal protein assembly sensitivity analysis statistical significance transgene virus resistance Adenocarcinoma Animals Antiviral Agents bcl-2-Associated X Protein beta-Galactosidase Bystander Effect Cell Division Cell Line, Tumor Drug Resistance Ganciclovir Gene Therapy Mammary Neoplasms, Animal Melanoma Mice Mice, Inbred BALB C Proto-Oncogene Proteins c-bcl-2 Simplexvirus Spheroids, Cellular Thymidine Kinase Transfection Viral Proteins Animalia Murinae Simplexvirus
description	We have developed multicellular spheroids (MCS) established from LM05e and LM3 spontaneous Balb/c-murine mammary adenocarcinoma and B16 C57-murine melanoma derived cell lines as an in vitro model to study the efficacy of the herpes simplex virus thymidine kinase/ganciclovir (HSVtk/GCV) suicide system. We demonstrated for the first time that HSVtk-expressing cells assembled as MCS manifested a GCV resistance phenotype compared to the same cells grown as sparse monolayers. HSVtk-expressing LM05e, LM3 and B16 spheroids were 16-, three- and nine-fold less sensitive to GCV than their respective monolayers, even though they could express transgenes 10-, eight- and five-fold more efficiently. Mixed populations of HSVtk- and their respective βgal-expressing cells displayed a cell-type specific bystander effect that was higher in monolayers than in MCS. However, HSVtk-expressing cells in two- or three-dimensional cultures were always significantly more sensitive to GCV than the βgal-expressing counterparts, supporting the feasibility of this suicide approach in vivo. We present evidence showing that HSVtk-expressing tumor cells, when transferred from monolayers to MCS, displayed: (i) lower GCV cytotoxic activity and bystander effect; (ii) higher and efficient expression of genes transferred as lipoplexes; (iii) lower cell proliferation rates; and (iv) changes in intracellular Bax/Bcl-xL rheostat of mitochondria-mediated apoptosis.
format	JOUR
author	Finocchiaro, L.M.E. Bumaschny, V.F. Karara, A.L. Fiszman, G.L. Casais, C.C. Glikin, G.C.
author_facet	Finocchiaro, L.M.E. Bumaschny, V.F. Karara, A.L. Fiszman, G.L. Casais, C.C. Glikin, G.C.
author_sort	Finocchiaro, L.M.E.
title	Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
title_short	Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
title_full	Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
title_fullStr	Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
title_full_unstemmed	Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
title_sort	herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids
url	http://hdl.handle.net/20.500.12110/paper_09291903_v11_n5_p333_Finocchiaro
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_version_	1807320200338472960

Herpes simplex virus thymidine kinase/ganciclovir system in multicellular tumor spheroids

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