Adenovirus vector-mediated delivery of the prodrug-converting enzyme carboxypeptidase G2 in a secreted or GPI-anchored form: High-level expression of this active conditional cytotoxic enzyme at the plasma membrane.

2.50
Hdl Handle:
http://hdl.handle.net/2436/29772
Title:
Adenovirus vector-mediated delivery of the prodrug-converting enzyme carboxypeptidase G2 in a secreted or GPI-anchored form: High-level expression of this active conditional cytotoxic enzyme at the plasma membrane.
Authors:
Cowen, Rachel L.; Williams, Judith C.; Emery, Steve; Blakey, David; Darling, John L.; Lowenstein, Pedro R.; Castro, Maria G.
Abstract:
Carboxypeptidase G2 (CPG2) is a powerful prodrug-converting enzyme. Without a requirement for endogenous enzymes or cofactors, it can directly activate mustard alkylating prodrugs to cytotoxic species, killing both quiescent and dividing cells. This paper provides the first report of its use in the context of a clinically relevant delivery vehicle using adenovirus vectors. To strengthen the efficacy of the prodrug-activating system, the enzyme has been engineered to be secreted or glycosylphosphatidylinositol (GPI) anchored to the extracellular membrane of tumor cells, resulting in an enhanced bystander effect by facilitating diffusion of the active drug through extracellular, rather than intracellular, activation. Using the vectors, we have achieved expression of functional secreted or GPI-anchored CPG2 in a panel of tumor cell lines demonstrating no loss in efficacy as a result of GPI anchor retention. Despite variable transduction efficiencies inherent to these vectors, greater than 50% cell kill was achievable in all of the cell lines tested following only a single exposure to the prodrug ZD2767P. Even in cell lines refractive to infection with the vectors, substantial cell death was recorded, indicative of the enhanced bystander effect generated following extracellular prodrug activation. A direct evaluation of the efficacy of our system has been made against adenoviral delivery of herpes simples virus thymidine kinase plus ganciclovir (GCV), a suicide gene therapy approach already in the clinic. In a short-term human glioma culture (IN1760) resistant to the clinical chemotherapeutic drug CCNU (1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea), thymidine kinase/GCV effected no cell killing compared to 70% cell killing with our system.
Citation:
Cancer Gene Therapy, 9 (11): 897-907
Publisher:
nature.com
Journal:
Cancer Gene Therapy
Issue Date:
2002
URI:
http://hdl.handle.net/2436/29772
DOI:
10.1038/sj.cgt.7700514
PubMed ID:
12386828
Additional Links:
http://www.nature.com/cgt/journal/v9/n11/abs/7700514a.html; http://direct.bl.uk/bld/PlaceOrder.do?UIN=122740146&ETOC=RN&from=searchengine
Type:
Article
Language:
en
ISSN:
0929-1903
Appears in Collections:
Cancer Research Group

Full metadata record

DC FieldValue Language
dc.contributor.authorCowen, Rachel L.-
dc.contributor.authorWilliams, Judith C.-
dc.contributor.authorEmery, Steve-
dc.contributor.authorBlakey, David-
dc.contributor.authorDarling, John L.-
dc.contributor.authorLowenstein, Pedro R.-
dc.contributor.authorCastro, Maria G.-
dc.date.accessioned2008-06-10T13:15:09Z-
dc.date.available2008-06-10T13:15:09Z-
dc.date.issued2002-
dc.identifier.citationCancer Gene Therapy, 9 (11): 897-907en
dc.identifier.issn0929-1903-
dc.identifier.pmid12386828-
dc.identifier.doi10.1038/sj.cgt.7700514-
dc.identifier.urihttp://hdl.handle.net/2436/29772-
dc.description.abstractCarboxypeptidase G2 (CPG2) is a powerful prodrug-converting enzyme. Without a requirement for endogenous enzymes or cofactors, it can directly activate mustard alkylating prodrugs to cytotoxic species, killing both quiescent and dividing cells. This paper provides the first report of its use in the context of a clinically relevant delivery vehicle using adenovirus vectors. To strengthen the efficacy of the prodrug-activating system, the enzyme has been engineered to be secreted or glycosylphosphatidylinositol (GPI) anchored to the extracellular membrane of tumor cells, resulting in an enhanced bystander effect by facilitating diffusion of the active drug through extracellular, rather than intracellular, activation. Using the vectors, we have achieved expression of functional secreted or GPI-anchored CPG2 in a panel of tumor cell lines demonstrating no loss in efficacy as a result of GPI anchor retention. Despite variable transduction efficiencies inherent to these vectors, greater than 50% cell kill was achievable in all of the cell lines tested following only a single exposure to the prodrug ZD2767P. Even in cell lines refractive to infection with the vectors, substantial cell death was recorded, indicative of the enhanced bystander effect generated following extracellular prodrug activation. A direct evaluation of the efficacy of our system has been made against adenoviral delivery of herpes simples virus thymidine kinase plus ganciclovir (GCV), a suicide gene therapy approach already in the clinic. In a short-term human glioma culture (IN1760) resistant to the clinical chemotherapeutic drug CCNU (1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea), thymidine kinase/GCV effected no cell killing compared to 70% cell killing with our system.en
dc.language.isoenen
dc.publishernature.comen
dc.relation.urlhttp://www.nature.com/cgt/journal/v9/n11/abs/7700514a.htmlen
dc.relation.urlhttp://direct.bl.uk/bld/PlaceOrder.do?UIN=122740146&ETOC=RN&from=searchengineen
dc.subjectSuicide gene therapyen
dc.subjectCarboxypeptidase G2en
dc.subjectChemosensitivityen
dc.subjectGPI anchoren
dc.subjectOncologyen
dc.subjectCancer treatmenten
dc.subject.meshAdenoviridaeen
dc.subject.meshAmino Acid Sequenceen
dc.subject.meshAnimalsen
dc.subject.meshAntigens, Thy-1en
dc.subject.meshBase Sequenceen
dc.subject.meshCell Membraneen
dc.subject.meshCell Survivalen
dc.subject.meshDNA Primersen
dc.subject.meshExonsen
dc.subject.meshGenetic Vectorsen
dc.subject.meshGlycosylphosphatidylinositolsen
dc.subject.meshHumansen
dc.subject.meshMolecular Sequence Dataen
dc.subject.meshMutagenesis, Site-Directeden
dc.subject.meshProdrugsen
dc.subject.meshRatsen
dc.subject.meshRecombinant Fusion Proteinsen
dc.subject.meshTransfectionen
dc.subject.meshTumor Cells, Cultureden
dc.subject.meshgamma-Glutamyl Hydrolaseen
dc.titleAdenovirus vector-mediated delivery of the prodrug-converting enzyme carboxypeptidase G2 in a secreted or GPI-anchored form: High-level expression of this active conditional cytotoxic enzyme at the plasma membrane.en
dc.typeArticleen
dc.identifier.journalCancer Gene Therapyen

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