Hepatic control elements promote long-term expression of human coagulation factor IX gene in hydrodynamically transfected mice

J Gene Med. 2011 Jul;13(7-8):365-72. doi: 10.1002/jgm.1583.

Abstract

Background: Long-term expression of the delivered target gene is critical for successful gene therapy. Recently, hepatic control region I (HCR I) originating from the apolipoprotein (apo)C-I pseudogene was shown to be a critical element for long-term gene expression in the liver of mice. HCR II is another hepatic control region of apoC-I.

Methods: HCR I, HCR II and HCR I/II-containing plasmids encoding factor IX were prepared and hydrodynamically transferred into the liver of normal and hemophilia B mice. Factor IX expression, clotting activity and formation of antibodies against the expressed gene product were compared.

Results: HCR I-, HCR II- and HCR I/II-containing plasmids all induced long-term gene expression in both normal and hemophilia B mice. Post-transfection factor IX expression in the hemophilia B mice remained above 500 ng/ml for 210 days. Antibodies against human factor IX were detected at a low level in the serum, although they had no effect on the levels and clotting activity of the expressed factor IX.

Conclusions: We have shown in mouse models that hydrodynamic transfection of pBS-HCRII-HP-FIXA and pBS-HCRI/II-HP-FIXA was able to induce and maintain the expression and clotting activity of human factor IX for a long period of time at a potentially therapeutic level. With an appropriate delivery system, this type of plasmid vector could be clinically useful for the hepatic expression of therapeutic genes including human factor IX.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Neutralizing / blood
  • Antibodies, Neutralizing / immunology
  • Cell Line, Tumor
  • Dependovirus / genetics
  • Disease Models, Animal
  • Factor IX / genetics*
  • Factor IX / immunology
  • Factor IX / metabolism*
  • Gene Expression Regulation
  • Gene Transfer Techniques
  • Genetic Therapy
  • Genetic Vectors / genetics
  • Hemophilia B / genetics*
  • Hemophilia B / metabolism*
  • Hemophilia B / therapy
  • Hep G2 Cells
  • Humans
  • Liver / metabolism*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Organ Specificity / genetics
  • Partial Thromboplastin Time
  • Promoter Regions, Genetic
  • Regulatory Elements, Transcriptional*

Substances

  • Antibodies, Neutralizing
  • Factor IX