title: Long-term therapeutic levels of human alpha-1 antitrypsin in plasma after hydrodynamic injection of nonviral DNA.
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.
authors.affiliation: Departamento de Farmacología, Facultad de Medicina, Universidad de Valencia, Valencia, Spain.
authors.forename: S F
authors.initials: SF
authors.lastname: Aliño
authors.forename: A
authors.initials: A
authors.lastname: Crespo
authors.forename: F
authors.initials: F
authors.lastname: Dasí
mesh_terms: D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000008:administration & dosage*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry; D007267:Injections; D008099:Liver / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D019076:Transgenes; D000515:alpha 1-Antitrypsin / Q000032:analysis* / Q000235:genetics*
chemical_list: D000515:alpha 1-Antitrypsin; D004247:DNA