Intrabiliary infusion of naked DNA vectors targets periportal hepatocytes in mice

Deplazes, Sereina, Schlegel, Andrea, Song, Zhuolun, Allegri, Gabriella, Rimann, Nicole, Scherer, Tanja, Willimann, Melanie, Opitz, Lennart, Cunningham, Sharon C, Alexander, Ian E
et al (show 4 more authors) (2022) Intrabiliary infusion of naked DNA vectors targets periportal hepatocytes in mice. MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT, 27. pp. 352-367.

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Hydrodynamic tail vein injection (HTV) is the "gold standard" for delivering naked DNA vectors to mouse liver, thereby transfecting predominately perivenous hepatocytes. While HTV corrects metabolic liver defects such as phenylketonuria or cystathionine β-synthase deficiency, correction of <i>spf</i> <sup><i>ash</i></sup> mice with ornithine transcarbamylase (OTC) deficiency was not possible despite overexpression in the liver, as the OTC enzyme is primarily expressed in periportal hepatocytes. To target periportal hepatocytes, we established hydrodynamic retrograde intrabiliary injection (HRII) in mice and optimized minicircle (MC) vector delivery using luciferase as a marker gene. HRII resulted in a transfection efficiency below 1%, 100-fold lower than HTV. While HRII induced minimal liver toxicity compared with HTV, overexpression of luciferase by both methods, but not of a natural liver-specific enzyme, elicited an immune response that led to the elimination of luciferase expression. Further testing of MC vectors delivered via HRII in <i>spf</i> <sup><i>ash</i></sup> mice did not result in sufficient therapeutic efficacy and needs further optimization and/or selection of the corrected cells. This study reveals that luciferase expression is toxic for the liver. Furthermore, physical delivery of MC vectors via the bile duct has the potential to treat defects restricted to periportal hepatocytes, which opens new doors for non-viral liver-directed gene therapy.

Item Type: Article
Uncontrolled Keywords: FLAG-tagged OTC, OTC deficiency, hydrodynamic retrograde intrabiliary injection, hydrodynamic tail vein injection, immune response, liver toxicity, liver-directed gene therapy, luciferase expression, naked DNA minicircle vector, non-viral, urea cycle disorder
Divisions: Faculty of Health and Life Sciences
Faculty of Health and Life Sciences > Institute of Infection, Veterinary and Ecological Sciences
Depositing User: Symplectic Admin
Date Deposited: 31 Jan 2023 12:47
Last Modified: 31 Jan 2023 12:47
DOI: 10.1016/j.omtm.2022.10.006
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