Luciferase-based reporting of suicide gene activity in murine mesenchymal stem cells

Due to their ease of isolation, gene modification and tumor-homing properties, mesenchymal stem cells (MSCs) are an attractive cellular vehicle for the delivery of toxic suicide genes to a variety of cancers in pre-clinical models. In addition, the incorporation of suicide genes in stem cell-derived...

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Bibliographic Details
Published in:PloS one 2019-07, Vol.14 (7), p.e0220013
Main Authors: Gerace, Dario, Martiniello-Wilks, Rosetta, Habib, Rosaline, Simpson, Ann Margaret
Format: Article
Language:English
Subjects:
Adenoviruses
Animals
Apoptosis
Biology and Life Sciences
Bioluminescence
Biomarkers
Bone marrow
Breast cancer
Cancer therapies
Cell Line
Cell migration
Chemotherapy
Cytosine
Cytosine deaminase
Cytotoxicity
E coli
Fibroblasts
Fireflies
Gene Expression
Gene therapy
Genes
Genes, Reporter
Genes, Transgenic, Suicide
Genetic aspects
Genetic modification
Genetically modified organisms
Homing
Immunophenotyping
Kinases
Life sciences
Luciferase
Luciferases - genetics
Luciferases - metabolism
Medical research
Medicine and Health Sciences
Mesenchymal stem cells
Mesenchymal Stem Cells - cytology
Mesenchymal Stem Cells - metabolism
Mesenchyme
Methylene blue
Mice
Phosphoribosyltransferase
Physical Sciences
Physiological aspects
Plasmids
Pyrimidines
Research and Analysis Methods
Stem cell transplantation
Stem cells
Suicidal behavior
Suicide
Suicide genes
Toxicity
Tumors
Uracil
Uracil phosphoribosyltransferase
Yeast
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Summary:Due to their ease of isolation, gene modification and tumor-homing properties, mesenchymal stem cells (MSCs) are an attractive cellular vehicle for the delivery of toxic suicide genes to a variety of cancers in pre-clinical models. In addition, the incorporation of suicide genes in stem cell-derived cell replacement therapies improves their safety profile by permitting graft destruction in the event of unexpected tumorigeneses or unwanted differentiation. Due to the functional requirement of ATP for the Firefly luciferase gene Luc2 to produce light, luciferase-based reporting of cytotoxicity can be engineered into potential cell therapies. Consequently, we nucleofected mammalian expression plasmids containing both the Luc2 and the yeast fusion cytosine deaminase uracil phosphoribosyltransferase (CDUPRT) genes for expression in murine MSCs to assess luciferase as a reporter of suicide gene cytotoxicity, and MSC as vehicles of suicide gene therapy. In vitro bioluminescence imaging (BLI) showed that following the addition of the non-toxic prodrug fluorocytosine (5-FC), CDUPRT-expressing MSCs displayed enhanced cytotoxicity in comparison to Luc2 reporter MSC controls. This study demonstrates the utility of luciferase as a reporter of CDUPRT-mediated cytotoxicity in murine MSC using BLI.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0220013