Ligand-dependent regulation of plasmid-based transgene expression in vivo

被引:47
作者
Abruzzese, RV
Godin, D
Burcin, M
Mehta, V
French, M
Li, YH
O'Malley, BW
Nordstrom, JL
机构
[1] Valentis Inc, The Woodlands, TX 77381 USA
[2] Baylor Coll Med, Dept Cell Biol, Houston, TX 77030 USA
关键词
D O I
10.1089/10430349950017833
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
As gene therapy advances, the ability to regulate transgene expression will become paramount for safety and efficacy, In this study, we investigate the ability of the mifepristone-dependent GeneSwitch system to regulate the expression of trangenes delivered to mice by nonviral methods. Two plasmids, one encoding the chimeric GeneSwitch protein, the other an inducible transgene for secreted human placental alkaline phosphatase (SEAP), were delivered to the hind-limb muscles of adult mice. Modulation of the level of secretion of the transgene product into serum was achieved by intraperitoneal administration of low doses of the drug mifepristone (MFP), The EC50 for induction of transgene expression by MFP was 0.03 +/- 0.005 mg/kg, The maximal level of transgene expression after induction was equal to or higher than that displayed by a plasmid driven by the CMV enhancer/promoter. The average magnitude of induction was 14- to 19-fold, Multiple rounds of drug-dependent regulation of transgene expression in vivo were demonstrated, In BALB/c mice, the ability to regulate transgene expression persisted for approximately 3 weeks, until the appearance of neutralizing antibodies to the secreted transgene product. In immune-deficient mice, the ability to repetitively regulate transgene expression persisted for at least 5 weeks. Although the dynamic range of regulation needs improvement, the plasmid-based GeneSwitch system has features that are attractive for gene therapy applications.
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页码:1499 / 1507
页数:9
相关论文
共 21 条
[1]   Systemic effect of human growth hormone after intramuscular injection of a single dose of a muscle-specific gene medicine [J].
Anwer, K ;
Shi, M ;
French, MF ;
Muller, SR ;
Chen, W ;
Liu, QS ;
Proctor, BL ;
Wang, JJ ;
Mumper, RJ ;
Singhal, A ;
Rolland, AP ;
Alila, HW .
HUMAN GENE THERAPY, 1998, 9 (05) :659-670
[2]   Control of erythropoietin delivery by doxycycline in mice after intramuscular injection of adeno-associated vector [J].
Bohl, D ;
Salvetti, A ;
Moullier, P ;
Heard, JM .
BLOOD, 1998, 92 (05) :1512-1517
[3]  
Bronstein I, 1996, CLIN CHEM, V42, P1542
[4]   Adenovirus-mediated regulable target gene expression in vivo [J].
Burcin, MM ;
Schiedner, G ;
Kochanek, S ;
Tsai, SY ;
O'Malley, BW .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (02) :355-360
[5]   TETRACYCLINE-REGULATED GENE-EXPRESSION FOLLOWING DIRECT GENE-TRANSFER INTO MOUSE SKELETAL-MUSCLE [J].
DHAWAN, J ;
RANDO, TA ;
ELSON, SL ;
BUJARD, H ;
BLAU, HM .
SOMATIC CELL AND MOLECULAR GENETICS, 1995, 21 (04) :233-240
[6]   Tetracycline-responsive gene expression in mouse brain after amplicon-mediated gene transfer [J].
Fotaki, ME ;
Pink, JR ;
Mous, J .
GENE THERAPY, 1997, 4 (09) :901-908
[7]   TREATMENT OF UNRESECTABLE MENINGIOMAS WITH THE ANTIPROGESTERONE AGENT MIFEPRISTONE [J].
GRUNBERG, SM ;
WEISS, MH ;
SPITZ, IM ;
AHMADI, J ;
SADUN, A ;
RUSSELL, CA ;
LUCCI, L ;
STEVENSON, LL .
JOURNAL OF NEUROSURGERY, 1991, 74 (06) :861-866
[8]  
HARDING TC, 1997, NAT BIOTECHNOL, V16, P5553
[9]   Inducible gene expression from defective herpes simplex virus vectors using the tetracycline-responsive promoter system [J].
Ho, DY ;
McLaughlin, JR ;
Sapolsky, RM .
MOLECULAR BRAIN RESEARCH, 1996, 41 (1-2) :200-209
[10]  
Levy MY, 1996, GENE THER, V3, P201