Non-invasive bioluminescence imaging of myoblast-mediated hypoxia-inducible factor-1 alpha gene transfer.
OBJECTIVE
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.


METHODS
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.
Molecular Imaging Program at Stanford (MIPS), Department of Radiology, Division of Nuclear Medicine, Stanford University, Stanford, CA 94305-5427, USA.
Olivier
O
Gheysens
Ian Y
IY
Chen
Martin
M
Rodriguez-Porcel
Carmel
C
Chan
Julia
J
Rasooly
Caroline
C
Vaerenberg
Ramasamy
R
Paulmurugan
Juergen K
JK
Willmann
Christophe
C
Deroose
Joseph
J
Wu
Sanjiv S
SS
Gambhir
D000818:Animals; D002452:Cell Count; D002460:Cell Line; D017690:Cell Transplantation; D057075:Enzyme Assays; D004797:Enzyme-Linked Immunosorbent Assay; D005240:Feasibility Studies; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008027:Light; D049410:Luciferases, Renilla / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D032446:Myoblasts / Q000378:metabolism*; D018919:Neovascularization, Physiologic; D010957:Plasmids / Q000235:genetics; D014162:Transfection; D019076:Transgenes / Q000235:genetics
D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D049410:Luciferases, Renilla
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