Low-Dose Gene Therapy for Murine PKU Using Episomal Naked DNA Vectors Expressing PAH from Its Endogenous Liver Promoter

Hiu Man Grisch-Chan, Andrea Schlegel, Tanja Scherer, Gabriella Allegri, Raphael Heidelberger, Panagiota Tsikrika, Marco Schmeer, Martin Schleef, Cary Harding, Johannes Häberle, Beat Thöny

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)339-349
Number of pages11
JournalMolecular Therapy - Nucleic Acids
Volume7
DOIs
StatePublished - Jun 1 2017

Fingerprint

Phenylalanine Hydroxylase
Phenylketonurias
Genetic Therapy
Liver
DNA
Transgenes
Genome
Insertional Mutagenesis
Hydrodynamics
Phenylalanine
Codon
Introns
Complementary DNA
Safety
Injections
Therapeutics

ASJC Scopus subject areas

  • Molecular Medicine
  • Drug Discovery

Cite this

Grisch-Chan, H. M., Schlegel, A., Scherer, T., Allegri, G., Heidelberger, R., Tsikrika, P., ... Thöny, B. (2017). Low-Dose Gene Therapy for Murine PKU Using Episomal Naked DNA Vectors Expressing PAH from Its Endogenous Liver Promoter. Molecular Therapy - Nucleic Acids, 7, 339-349. https://doi.org/10.1016/j.omtn.2017.04.013

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; Häberle, Johannes; Thöny, Beat.

In: Molecular Therapy - Nucleic Acids, Vol. 7, 01.06.2017, p. 339-349.

Research output: Contribution to journalArticle

Grisch-Chan, HM, Schlegel, A, Scherer, T, Allegri, G, Heidelberger, R, Tsikrika, P, Schmeer, M, Schleef, M, Harding, C, Häberle, J & Thöny, B 2017, 'Low-Dose Gene Therapy for Murine PKU Using Episomal Naked DNA Vectors Expressing PAH from Its Endogenous Liver Promoter', Molecular Therapy - Nucleic Acids, vol. 7, pp. 339-349. https://doi.org/10.1016/j.omtn.2017.04.013
Grisch-Chan, Hiu Man ; Schlegel, Andrea ; Scherer, Tanja ; Allegri, Gabriella ; Heidelberger, Raphael ; Tsikrika, Panagiota ; Schmeer, Marco ; Schleef, Martin ; Harding, Cary ; Häberle, Johannes ; Thöny, Beat. / Low-Dose Gene Therapy for Murine PKU Using Episomal Naked DNA Vectors Expressing PAH from Its Endogenous Liver Promoter. In: Molecular Therapy - Nucleic Acids. 2017 ; Vol. 7. pp. 339-349.
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