733. Enhancement of Lentiviral-Mediated Gene Transfer in Kidney by Polycations and Detergents

• Published in: Molecular therapy Vol. 9; no. S1; p. S279
• Main Authors: Fedorova, Elena V, Battini, Lorenzo, Hanss, Basil, Gusella, G Luca
• Format: Journal Article
• Language: English
• Published: Milwaukee Elsevier Inc 01.05.2004; Elsevier Limited
• Subjects: Cloning; Efficiency; Gene therapy; Kidneys; Leukemia; Packaging; Peptides; Vectors (Biology); Denmark; Aarhus Denmark
• ISSN: 1525-0016; 1525-0024
• DOI: 10.1016/j.ymthe.2004.06.674

We have previously shown that stable transduction of kidney can be achieved by the delivery of lentiviral vectors through the ureter and to the renal parenchyma. In this report, we utilized the same routes of delivery to assess the ability of different agents to enhance viral transduction of kidney. In particular, we focused on mannitol, a hyperosmotic agent, polybrene, a polycation that increases virus adsorption to cell surface, as well as the mild detergents polidocanol and lysophosphatidylcholine, which have been reported to improve the efficiency of transduction in a variety of epithelial tissues. These agents were tested in Balb/c mice during the ureter or intraparenchymal administration of the third generation replication-defective, self-inactivating lentiviral vectors (VVCW or VVPW) carrying EGFP, LacZ, or SEAP as reporter genes under the control of the eCMV or murine PGK promoters. Approximately 1×10 7 to 1×108 transducing units (TU) of the lentiviral vector were formulated in 50 μl of mannitol or polybrene solutions before delivery. Alternatively, either polidocanol or lysophosphatidylcholine was used to pretreat the kidneys for one hour prior to infusion of 10 8 viral TU. Transgene expression was then evaluated two weeks later by determining the expression of the reporter gene in renal cells, urine, or blood. Positive transduction of the mouse kidney was confirmed by immunohistochemical staining of the kidney for bacterial β-galactosidase or EGFP and by detection of SEAP activity in kidney extracts. Kidney transduction was observed with virus alone independently of the treatment with any of the agents and was used as control baseline. Mannitol did not affect the levels of viral transduction. However, the presence of polybrene or the pretreatment with polidocanol or lysophosphatidylcholine enhanced the lentiviral mediated gene transfer to kidney. When the lentivector preparation was centrifuged in the presence of polybrene and administered through the ureter and parenchyma, transduction efficiency increased seven- and two-fold, respectively. Retrograde ureter infusion in kidney following pretreatment with PDOC or LPC resulted in a six- or two-fold increase in transduction levels, respectively. The positive effect of these agents on gene transfer appeared not to be associated with any histological abnormality or with obvious inflammatory response within the mouse kidney.The identification of polybrene, polidocanol, and lysophosphatidylcholine as adjuvant of lentiviral mediated gene transduction of kidney underlies the potential of lentiviral vectors for the treatment of kidney diseases and for the development of models to study renal physiology. These results also suggest that these compounds may enhance the lentiviral gene transfer to other epithelial tissues.