Treatment of phenylketonuria using minicircle-based naked-DNA gene transfer to murine liver

Hiu Man Viecelli, Richard P. Harbottle, Suet Ping Wong, Andrea Schlegel, Marinee K. Chuah, Thierry Vandendriessche, Cary Harding, Beat Thöny

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Host immune response to viral vectors, persistence of nonintegrating vectors, and sustained transgene expression are among the major challenges in gene therapy. To overcome these hurdles, we successfully used minicircle (MC) naked-DNA vectors devoid of any viral or bacterial sequences for the long-term treatment of murine phenylketonuria, a model for a genetic liver defect. MC-DNA vectors expressed the murine phenylalanine hydroxylase (Pah) complementary DNA (cDNA) from a liver-specific promoter coupled to a de novo designed hepatocyte-specific regulatory element, designated P3, which is a cluster of evolutionary conserved transcription factor binding sites. MC-DNA vectors were subsequently delivered to the liver by a single hydrodynamic tail vein (HTV) injection. The MC-DNA vector normalized blood phenylalanine concomitant with reversion of hypopigmentation in a dose-dependent manner for more than 1 year, whereas the corresponding parental plasmid did not result in any phenylalanine clearance. MC vectors persisted in an episomal state in the liver consistent with sustained transgene expression and hepatic PAH enzyme activity without any apparent adverse effects. Moreover, 14-20% of all hepatocytes expressed transgenic PAH, and the expression was observed exclusively in the liver and predominately around pericentral areas of the hepatic lobule, while there was no transgene expression in periportal areas. Conclusion: This study demonstrates that MC technology offers an improved safety profile and has the potential for the genetic treatment of liver diseases.

Original languageEnglish (US)
Pages (from-to)1035-1043
Number of pages9
JournalHepatology
Volume60
Issue number3
DOIs
StatePublished - 2014

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Phenylketonurias
Liver
DNA
Genes
Transgenes
Phenylalanine
Therapeutics
Hepatocytes
Phenylalanine Hydroxylase
Hypopigmentation
Genetic Models
Hydrodynamics
Genetic Therapy
Liver Diseases
Veins
Plasmids
Transcription Factors
Complementary DNA
Binding Sites
Technology

ASJC Scopus subject areas

  • Hepatology

Cite this

Viecelli, H. M., Harbottle, R. P., Wong, S. P., Schlegel, A., Chuah, M. K., Vandendriessche, T., ... Thöny, B. (2014). Treatment of phenylketonuria using minicircle-based naked-DNA gene transfer to murine liver. Hepatology, 60(3), 1035-1043. https://doi.org/10.1002/hep.27104

Treatment of phenylketonuria using minicircle-based naked-DNA gene transfer to murine liver. / Viecelli, Hiu Man; Harbottle, Richard P.; Wong, Suet Ping; Schlegel, Andrea; Chuah, Marinee K.; Vandendriessche, Thierry; Harding, Cary; Thöny, Beat.

In: Hepatology, Vol. 60, No. 3, 2014, p. 1035-1043.

Research output: Contribution to journalArticle

Viecelli, HM, Harbottle, RP, Wong, SP, Schlegel, A, Chuah, MK, Vandendriessche, T, Harding, C & Thöny, B 2014, 'Treatment of phenylketonuria using minicircle-based naked-DNA gene transfer to murine liver', Hepatology, vol. 60, no. 3, pp. 1035-1043. https://doi.org/10.1002/hep.27104
Viecelli HM, Harbottle RP, Wong SP, Schlegel A, Chuah MK, Vandendriessche T et al. Treatment of phenylketonuria using minicircle-based naked-DNA gene transfer to murine liver. Hepatology. 2014;60(3):1035-1043. https://doi.org/10.1002/hep.27104
Viecelli, Hiu Man ; Harbottle, Richard P. ; Wong, Suet Ping ; Schlegel, Andrea ; Chuah, Marinee K. ; Vandendriessche, Thierry ; Harding, Cary ; Thöny, Beat. / Treatment of phenylketonuria using minicircle-based naked-DNA gene transfer to murine liver. In: Hepatology. 2014 ; Vol. 60, No. 3. pp. 1035-1043.
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