TY - JOUR
T1 - Repetitive Imaging of Reporter Gene Expression in the Lung
AU - Richard, Jean Christophe
AU - Factor, Philip
AU - Ferkol, Thomas
AU - Ponde, Datta E.
AU - Zhou, Zhaohui
AU - Schuster, Daniel P.
PY - 2003/10
Y1 - 2003/10
N2 - Positron emission tomographic imaging is emerging as a powerful technology to monitor reporter transgene expression in the lungs and other organs. However, little information is available about its usefulness for studying gene expression over time. Therefore, we infected 20 rats with a replication-deficient adenovirus containing a fusion gene encoding for a mutant Herpes simplex virus type-1 thymidine kinase and an enhanced green fluorescent protein. Five additional rats were infected with a control virus. Pulmonary gene transfer was performed via intratracheal administration of vector using a surfactant-based method. Imaging was performed 4-6 hr, and 4, 7, and 10 days after gene transfer, using 9-(4-[ 18F]-fluoro-3-hydroxyme-thylbutyl)guanine, an imaging substrate for the mutant kinase. Lung tracer uptake assessed with imaging was moderately but significantly increased 4-6 hr after gene transfer, was maximal after 4 days, and was no longer detectable by 10 days. The temporal pattern of transgene expression measured ex vivo with in vitro assays of thymidine kinase activity and green fluorescent protein was similar to imaging. In conclusion, positron emission tomography is a reliable new tool to evaluate the onset and duration of reporter gene expression non-invasively in the lungs of intact animals.
AB - Positron emission tomographic imaging is emerging as a powerful technology to monitor reporter transgene expression in the lungs and other organs. However, little information is available about its usefulness for studying gene expression over time. Therefore, we infected 20 rats with a replication-deficient adenovirus containing a fusion gene encoding for a mutant Herpes simplex virus type-1 thymidine kinase and an enhanced green fluorescent protein. Five additional rats were infected with a control virus. Pulmonary gene transfer was performed via intratracheal administration of vector using a surfactant-based method. Imaging was performed 4-6 hr, and 4, 7, and 10 days after gene transfer, using 9-(4-[ 18F]-fluoro-3-hydroxyme-thylbutyl)guanine, an imaging substrate for the mutant kinase. Lung tracer uptake assessed with imaging was moderately but significantly increased 4-6 hr after gene transfer, was maximal after 4 days, and was no longer detectable by 10 days. The temporal pattern of transgene expression measured ex vivo with in vitro assays of thymidine kinase activity and green fluorescent protein was similar to imaging. In conclusion, positron emission tomography is a reliable new tool to evaluate the onset and duration of reporter gene expression non-invasively in the lungs of intact animals.
KW - FHBG
KW - Green fluorescent protein
KW - Herpes simplex virus 1 thymidine kinase
KW - Positron emission tomography
KW - Reporter gene
UR - http://www.scopus.com/inward/record.url?scp=1642434422&partnerID=8YFLogxK
U2 - 10.1162/153535003322750682
DO - 10.1162/153535003322750682
M3 - Article
C2 - 14717333
AN - SCOPUS:1642434422
SN - 1535-3508
VL - 2
SP - 342
EP - 349
JO - Molecular Imaging
JF - Molecular Imaging
IS - 4
ER -