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Intrabiliary infusion of naked DNA vectors targets periportal hepatocytes in mice
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 spf(ash)...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9630613/ https://www.ncbi.nlm.nih.gov/pubmed/36381301 http://dx.doi.org/10.1016/j.omtm.2022.10.006 |
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author | Deplazes, Sereina Schlegel, Andrea Song, Zhuolun Allegri, Gabriella Rimann, Nicole Scherer, Tanja Willimann, Melanie Opitz, Lennart Cunningham, Sharon C. Alexander, Ian E. Kipar, Anja Häberle, Johannes Thöny, Beat Grisch-Chan, Hiu Man |
author_facet | Deplazes, Sereina Schlegel, Andrea Song, Zhuolun Allegri, Gabriella Rimann, Nicole Scherer, Tanja Willimann, Melanie Opitz, Lennart Cunningham, Sharon C. Alexander, Ian E. Kipar, Anja Häberle, Johannes Thöny, Beat Grisch-Chan, Hiu Man |
author_sort | Deplazes, Sereina |
collection | PubMed |
description | 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 spf(ash) 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 spf(ash) 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. |
format | Online Article Text |
id | pubmed-9630613 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-96306132022-11-14 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. Kipar, Anja Häberle, Johannes Thöny, Beat Grisch-Chan, Hiu Man Mol Ther Methods Clin Dev Original Article 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 spf(ash) 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 spf(ash) 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. American Society of Gene & Cell Therapy 2022-10-10 /pmc/articles/PMC9630613/ /pubmed/36381301 http://dx.doi.org/10.1016/j.omtm.2022.10.006 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Article Deplazes, Sereina Schlegel, Andrea Song, Zhuolun Allegri, Gabriella Rimann, Nicole Scherer, Tanja Willimann, Melanie Opitz, Lennart Cunningham, Sharon C. Alexander, Ian E. Kipar, Anja Häberle, Johannes Thöny, Beat Grisch-Chan, Hiu Man Intrabiliary infusion of naked DNA vectors targets periportal hepatocytes in mice |
title | Intrabiliary infusion of naked DNA vectors targets periportal hepatocytes in mice |
title_full | Intrabiliary infusion of naked DNA vectors targets periportal hepatocytes in mice |
title_fullStr | Intrabiliary infusion of naked DNA vectors targets periportal hepatocytes in mice |
title_full_unstemmed | Intrabiliary infusion of naked DNA vectors targets periportal hepatocytes in mice |
title_short | Intrabiliary infusion of naked DNA vectors targets periportal hepatocytes in mice |
title_sort | intrabiliary infusion of naked dna vectors targets periportal hepatocytes in mice |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9630613/ https://www.ncbi.nlm.nih.gov/pubmed/36381301 http://dx.doi.org/10.1016/j.omtm.2022.10.006 |
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