Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes. Gene Ther 2013 Jan;20(1):24-34
Date
01/14/2012Pubmed ID
22241175DOI
10.1038/gt.2011.210Scopus ID
2-s2.0-84871978590 (requires institutional sign-in at Scopus site) 17 CitationsAbstract
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.
Author List
Scaife M, Pacienza N, Au BC, Wang JC, Devine S, Scheid E, Lee CJ, Lopez-Perez O, Neschadim A, Fowler DH, Foley R, Medin JAAuthor
Jeffrey A. Medin PhD Professor in the Pediatrics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsAntigens, CD19
Cell Death
Cell Line, Tumor
Fabry Disease
Genetic Vectors
HEK293 Cells
Humans
Lentivirus
Leukemia-Lymphoma, Adult T-Cell
Mice
Mice, Inbred NOD
Mice, SCID
Nucleoside-Phosphate Kinase
Protein Engineering
Receptor, Nerve Growth Factor
Recombinant Fusion Proteins
Transformation, Genetic
Zidovudine
