Fetal gene transfer using lentiviral vectors: in vivo detection of gene expression by microPET and optical imaging in fetal and infant monkeys.
Fetal intraperitoneal administration of human immunodeficiency virus (HIV)-l-derived lentiviral vectors (10(7) infectious particles/fetus) has consistently shown high levels of transduction and gene expression in the omentum, peritoneum, and diaphragm when assessed by polymerase chain reaction (PCR) and whole tissue fluorescence. In vivo imaging techniques were explored with early-gestation long-tailed macaques that were administered the vesicular stomatitis virus-glycoprotein (VSV-G)-pseudotyped HIV-1-derived lentiviral vector expressing a mutant herpes simplex virus type 1 thymidine kinase (HSV-1-sr39tk) and firefly luciferase under the control of the cytomegalovirus (CMV) promoter. Fetuses were monitored sonographically and twice during gestation 9-[4-[18F]Fluoro-3-(hydroxymethyl)butyl]guanine (18F-FHBG) was injected into the fetal circulation under ultrasound guidance in preparation for microPET imaging. All newborns were delivered at term by cesarean section and raised in the nursery for postnatal studies. At 2 months postnatal age, animals were imaged and biodistribution was assessed. Optical imaging for firefly luciferase expression was also performed every 2 months postnatal age. Under all imaging conditions gene expression was observed in the abdominal region, and closely paralleled findings from prior studies based on whole tissue fluorescence. These investigations have shown that HSV-1-sr39tk and firefly luciferase can be used to safely detect transgene expression at multiple time points in fetal and infant monkeys in vivo and without evidence of adverse effects.
Tarantal
Alice F
AF
Center for Fetal Monkey Gene Transfer for Heart, Lung, and Blood Diseases, University of California, Davis, Davis, CA 95616, USA. aftarantal@primate.ucdavis.edu
Lee
C Chang I
CC
Jimenez
Daniel F
DF
Cherry
Simon R
SR
D000818:Animals; D000831:Animals, Newborn; D005260:Female; D046128:Fetal Therapies / Q000379:methods*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics; D018259:Herpesvirus 1, Human / Q000235:genetics; D016086:Lentivirus / Q000235:genetics*; D049409:Luciferases, Firefly / Q000235:genetics; D008252:Macaca fascicularis; D008562:Membrane Glycoproteins / Q000235:genetics; D055095:Optics and Photonics; D049268:Positron-Emission Tomography; D011247:Pregnancy; D013937:Thymidine Kinase / Q000235:genetics; D014759:Viral Envelope Proteins / Q000235:genetics
C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D049409:Luciferases, Firefly; D013937:Thymidine Kinase