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Low-Dose Gene Therapy for Murine PKU Using Episomal Naked DNA Vectors Expressing PAH from Its Endogenous Liver Promoter
Limited duration of transgene expression, insertional mutagenesis, and size limitations for transgene cassettes pose challenges and risk factors for many gene therapy vectors. Here, we report on physiological expression of liver phenylalanine hydroxylase (PAH) by delivery of naked DNA/minicircle (MC...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5423318/ https://www.ncbi.nlm.nih.gov/pubmed/28624210 http://dx.doi.org/10.1016/j.omtn.2017.04.013 |
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author | Grisch-Chan, Hiu Man Schlegel, Andrea Scherer, Tanja Allegri, Gabriella Heidelberger, Raphael Tsikrika, Panagiota Schmeer, Marco Schleef, Martin Harding, Cary O. Häberle, Johannes Thöny, Beat |
author_facet | Grisch-Chan, Hiu Man Schlegel, Andrea Scherer, Tanja Allegri, Gabriella Heidelberger, Raphael Tsikrika, Panagiota Schmeer, Marco Schleef, Martin Harding, Cary O. Häberle, Johannes Thöny, Beat |
author_sort | Grisch-Chan, Hiu Man |
collection | PubMed |
description | Limited duration of transgene expression, insertional mutagenesis, and size limitations for transgene cassettes pose challenges and risk factors for many gene therapy vectors. Here, we report on physiological expression of liver phenylalanine hydroxylase (PAH) by delivery of naked DNA/minicircle (MC)-based vectors for correction of homozygous enu2 mice, a model of human phenylketonuria (PKU). Because MC vectors lack a defined size limit, we constructed a MC vector expressing a codon-optimized murine Pah cDNA that includes a truncated intron and is under the transcriptional control of a 3.6-kb native Pah promoter/enhancer sequence. This vector, delivered via hydrodynamic injection, yielded therapeutic liver PAH activity and sustained correction of blood phenylalanine comparable to viral or synthetic liver promoters. Therapeutic efficacy was seen with vector copy numbers of <1 vector genome per diploid hepatocyte genome and was achieved at a vector dose that was significantly lowered. Partial hepatectomy and subsequent liver regeneration was associated with >95% loss of vector genomes and PAH activity in liver, demonstrating that MC vectors had not integrated into the liver genome. In conclusion, MC vectors, which do not have a defined size-limitation, offer a favorable safety profile for hepatic gene therapy due to their non-integration in combination with native promoters. |
format | Online Article Text |
id | pubmed-5423318 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-54233182017-05-11 Low-Dose Gene Therapy for Murine PKU Using Episomal Naked DNA Vectors Expressing PAH from Its Endogenous Liver Promoter Grisch-Chan, Hiu Man Schlegel, Andrea Scherer, Tanja Allegri, Gabriella Heidelberger, Raphael Tsikrika, Panagiota Schmeer, Marco Schleef, Martin Harding, Cary O. Häberle, Johannes Thöny, Beat Mol Ther Nucleic Acids Original Article Limited duration of transgene expression, insertional mutagenesis, and size limitations for transgene cassettes pose challenges and risk factors for many gene therapy vectors. Here, we report on physiological expression of liver phenylalanine hydroxylase (PAH) by delivery of naked DNA/minicircle (MC)-based vectors for correction of homozygous enu2 mice, a model of human phenylketonuria (PKU). Because MC vectors lack a defined size limit, we constructed a MC vector expressing a codon-optimized murine Pah cDNA that includes a truncated intron and is under the transcriptional control of a 3.6-kb native Pah promoter/enhancer sequence. This vector, delivered via hydrodynamic injection, yielded therapeutic liver PAH activity and sustained correction of blood phenylalanine comparable to viral or synthetic liver promoters. Therapeutic efficacy was seen with vector copy numbers of <1 vector genome per diploid hepatocyte genome and was achieved at a vector dose that was significantly lowered. Partial hepatectomy and subsequent liver regeneration was associated with >95% loss of vector genomes and PAH activity in liver, demonstrating that MC vectors had not integrated into the liver genome. In conclusion, MC vectors, which do not have a defined size-limitation, offer a favorable safety profile for hepatic gene therapy due to their non-integration in combination with native promoters. American Society of Gene & Cell Therapy 2017-04-20 /pmc/articles/PMC5423318/ /pubmed/28624210 http://dx.doi.org/10.1016/j.omtn.2017.04.013 Text en © 2017 The Authors http://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 Grisch-Chan, Hiu Man Schlegel, Andrea Scherer, Tanja Allegri, Gabriella Heidelberger, Raphael Tsikrika, Panagiota Schmeer, Marco Schleef, Martin Harding, Cary O. Häberle, Johannes Thöny, Beat Low-Dose Gene Therapy for Murine PKU Using Episomal Naked DNA Vectors Expressing PAH from Its Endogenous Liver Promoter |
title | Low-Dose Gene Therapy for Murine PKU Using Episomal Naked DNA Vectors Expressing PAH from Its Endogenous Liver Promoter |
title_full | Low-Dose Gene Therapy for Murine PKU Using Episomal Naked DNA Vectors Expressing PAH from Its Endogenous Liver Promoter |
title_fullStr | Low-Dose Gene Therapy for Murine PKU Using Episomal Naked DNA Vectors Expressing PAH from Its Endogenous Liver Promoter |
title_full_unstemmed | Low-Dose Gene Therapy for Murine PKU Using Episomal Naked DNA Vectors Expressing PAH from Its Endogenous Liver Promoter |
title_short | Low-Dose Gene Therapy for Murine PKU Using Episomal Naked DNA Vectors Expressing PAH from Its Endogenous Liver Promoter |
title_sort | low-dose gene therapy for murine pku using episomal naked dna vectors expressing pah from its endogenous liver promoter |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5423318/ https://www.ncbi.nlm.nih.gov/pubmed/28624210 http://dx.doi.org/10.1016/j.omtn.2017.04.013 |
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