TY - JOUR
T1 - Non-invasive bioluminescence imaging of myoblast-mediated hypoxia-inducible factor-1 alpha gene transfer
AU - Gheysens, Olivier
AU - Chen, Ian Y.
AU - Rodriguez-Porcel, Martin
AU - Chan, Carmel
AU - Rasooly, Julia
AU - Vaerenberg, Caroline
AU - Paulmurugan, Ramasamy
AU - Willmann, Juergen K.
AU - Deroose, Christophe
AU - Wu, Joseph
AU - Gambhir, Sanjiv S.
N1 - Funding Information:
Acknowledgments. This work was supported in part by grants from the NCI ICMIC P50CA114747 (S.S.G.) and Fund for Scientific Research Belgium-Flanders (O.G.).
PY - 2011/12
Y1 - 2011/12
N2 - Purpose: We tested a novel imaging strategy, in which both the survival of transplanted myoblasts and their therapeutic transgene expression, a recombinant hypoxia-inducible factor- 1α (HIF-1α-VP2), can be monitored using firefly luciferase (fluc) and Renilla luciferase (hrl) bioluminescence reporter genes, respectively. Procedures: The plasmid pUbi-hrl-pUbi-HIF-1α-VP2, which expresses both hrl and HIF-1α-VP2 using two ubiquitin promoters, was characterized in vitro. C2c12 myoblasts stably expressing fluc and transiently transfected with pUbi-hrl-pUbi-HIF-1α-VP2 were injected into the mouse hindlimb. Both hrl and fluc expression were monitored using bioluminescence imaging (BLI). Results: Strong correlations existed between the expression of hRL and each of HIF-1α-VP2, VEGF, and PlGF (r 2>0.83, r 2>0.82, and r 2>0.97, respectively). In vivo, both transplanted cells and HIF-1α-VP2 transgene expression were successfully imaged using BLI. Conclusions: An objective evaluation of myoblast-mediated gene transfer in living mice can be performed by monitoring both the survival and the transgene expression of transplanted myoblasts using the techniques developed herein.
AB - Purpose: We tested a novel imaging strategy, in which both the survival of transplanted myoblasts and their therapeutic transgene expression, a recombinant hypoxia-inducible factor- 1α (HIF-1α-VP2), can be monitored using firefly luciferase (fluc) and Renilla luciferase (hrl) bioluminescence reporter genes, respectively. Procedures: The plasmid pUbi-hrl-pUbi-HIF-1α-VP2, which expresses both hrl and HIF-1α-VP2 using two ubiquitin promoters, was characterized in vitro. C2c12 myoblasts stably expressing fluc and transiently transfected with pUbi-hrl-pUbi-HIF-1α-VP2 were injected into the mouse hindlimb. Both hrl and fluc expression were monitored using bioluminescence imaging (BLI). Results: Strong correlations existed between the expression of hRL and each of HIF-1α-VP2, VEGF, and PlGF (r 2>0.83, r 2>0.82, and r 2>0.97, respectively). In vivo, both transplanted cells and HIF-1α-VP2 transgene expression were successfully imaged using BLI. Conclusions: An objective evaluation of myoblast-mediated gene transfer in living mice can be performed by monitoring both the survival and the transgene expression of transplanted myoblasts using the techniques developed herein.
KW - Bioluminescence imaging
KW - Cell-mediated gene transfer
KW - Hypoxia-inducible factor-1α
KW - Molecular imaging
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U2 - 10.1007/s11307-011-0471-9
DO - 10.1007/s11307-011-0471-9
M3 - Article
C2 - 21267661
AN - SCOPUS:84855699336
SN - 1536-1632
VL - 13
SP - 1124
EP - 1132
JO - Molecular Imaging and Biology
JF - Molecular Imaging and Biology
IS - 6
ER -