Long-term therapeutic levels of human alpha-1 antitrypsin in plasma after hydrodynamic injection of nonviral DNA

Gene Ther. 2003 Sep;10(19):1672-9. doi: 10.1038/sj.gt.3302065.

Abstract

The transfection efficacy of several vectors containing the full genomic hAAT gene with its natural promoter (pTG7101) and others containing the cDNA of hAAT gene driven by cytomegalovirus immediate-early promoter or the 0.5 kb upstream of hAAT gene sequence has been studied by hydrodynamic tail-vein injection (20 microg/mouse). pTG7101 (but not the other plasmids) results in therapeutic and stable concentration of hAAT in plasma. A dose-response study with this plasmid (0.3-320 microg/mouse) confirms that hAAT remains long-term stable in plasma, with therapeutic concentrations of hAAT (>0.9 mg/ml). The parameters of the dose-response curve were: R: 0.98, E(max) 3449.0+/- 279.7 microg/ml and EC(50) 1.2 x 10(12) plasmid-gene units. In addition, 4 months after transfection, the intrinsic efficacy of transgenic expression (amount of RNA/DNA) in mouse liver was 50-80% that normally expressed by the mouse gene. The important efficacy of nonviral genomic DNA opens a new avenue in the safety applications of human gene therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • DNA / administration & dosage*
  • Gene Expression
  • Genetic Therapy / methods*
  • Humans
  • Immunohistochemistry
  • Injections
  • Liver / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Reverse Transcriptase Polymerase Chain Reaction
  • Time Factors
  • Transfection / methods*
  • Transgenes
  • alpha 1-Antitrypsin / analysis*
  • alpha 1-Antitrypsin / genetics*

Substances

  • alpha 1-Antitrypsin
  • DNA

Associated data

  • GENBANK/K02212
  • GENBANK/M12585
  • GENBANK/X01683