DEVELOPMENT OF NOVEL APPROACH FOR GENE TRANSFER OF RODENT FETAL LUNG

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Introduction In utero gene therapy might have potential interest for treatment of congenital lung diseases. Aim To develop a new method for accurate injections into specific compartments of the rodent fetuses during early stages of lung development and to assess transduction efficiency of adenoviral and lentiviral vectors after in utero injection. Methods Guided by a 40 mHz ultrasound scanhead we performed intra-amniotic, intra-parenchymal and intra-pleural injections in rodent fetuses (n=131) at E15.5–E16.5. For injection, we used PBS, lentivirus and adenovirus with green fluorescent protein (GFP) as a reporter gene. We evaluated survival rate, morphometric parameters and pattern of viral transfection at E16.5; E18.5; E21.5 and P7. Results Dramatically different patterns of GFP expression were observed ranging from visceral and parietal pleural expression, to focal intraparenchymal expression, to diffuse airway and alveolar expression. Adenoviral vectors appeared more efficient than lentiviral vectors for lung transduction. Conclusions We describe a new model of image guided microinjection of viral vectors into rodent fetuses targeting the fetal lung as early as pseudoglandular stage. These results should facilitate studies of fetal lung gene therapy as well as the effect of growth factors on lung development.