Improved neuronal transgene expression from an AAV-2 vector with a hybrid CMV enhancer/PDGF-β promoter

被引:18
作者
Wang, CY
Guo, HY
Lim, TM
Ng, YK
Neo, HP
Hwang, PYK
Yee, WC
Wang, S
机构
[1] Natl Univ Singapore, Dept Biol Sci, Singapore 117543, Singapore
[2] Natl Univ Singapore, Inst Bioengn & Nanotechnol, Singapore 117543, Singapore
[3] Natl Univ Singapore, Natl Inst Neurosci, Singapore 117543, Singapore
[4] Natl Univ Singapore, Dept Anat, Singapore 117543, Singapore
关键词
adeno-associated virus; PDGF-beta promoter; CMV enhancer; neuron; gene expression;
D O I
10.1002/jgm.742
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
Background Adeno-associated virus type 2 (AAV-2) vectors are highly promising tools for gene therapy of neurological disorders. After accommodating a cellular promoter, AAV-2 vectors are able to drive sustained expression of transgene in the brain. This study aimed to develop AAV-2 vectors that also facilitate a high level of neuronal expression by enhancing the strength of a neuron-specific promoter, the human platelet-derived growth factor beta-chain (PDGF) promoter. Methods and results A hybrid promoter approach was adopted to fuse the enhancer of human cytomegalovirus immediately early (CMV) promoter to the PDGF promoter. In cultured cortex neurons, AAV-2 vectors containing the hybrid promoter augmented transgene expression up to 20-fold over that mediated by titer-matched AAV-2 vectors with the PDGF promoter alone and 4-fold over the CMV enhancer/promoter. Injection of AAV-2 vectors with the hybrid promoter into the rat striatum resulted in neuron-specific transgene expression, the level of which was about 10-fold higher than those provided by the two control AAV-2 expression cassettes at 4 weeks post-injection and maintained for at least 12 weeks. Gene expression in the substantia nigra through possible retrograde transport of the AAV-2 vectors injected into the striatum was not obvious. After direct injection of AAV-2 vectors into the transgene expression driven by the hybrid promoter was substantia nigra, observed specifically in dopaminergic neurons and its level was about 3 and 17 times higher than that provided by the PDGF promoter alone and the CMV enhancer/promoter, respectively. Conclusions Enhanced transgene capacity plus neuron-specificity of the AAV-2 vectors developed in this study might prove valuable for gene therapy of Parkinson's disease. Copyright (c) 2005 John Wiley & Sons, Ltd.
引用
收藏
页码:945 / 955
页数:11
相关论文
共 42 条
[31]   EFFICIENT AND SUSTAINED GENE-EXPRESSION IN PRIMARY T-LYMPHOCYTES AND PRIMARY AND CULTURED TUMOR-CELLS MEDIATED BY ADENOASSOCIATED VIRUS PLASMID DNA COMPLEXED TO CATIONIC LIPOSOMES [J].
PHILIP, R ;
BRUNETTE, E ;
KILINSKI, L ;
MURUGESH, D ;
MCNALLY, MA ;
UCAR, K ;
ROSENBLATT, J ;
OKARMA, TB ;
LEBKOWSKI, JS .
MOLECULAR AND CELLULAR BIOLOGY, 1994, 14 (04) :2411-2418
[32]   Inactivation of the very strong HCMV immediate early promoter by DNA CpG methylation in vitro [J].
Prosch, S ;
Stein, J ;
Staak, K ;
Liebenthal, C ;
Volk, HD ;
Kruger, DH .
BIOLOGICAL CHEMISTRY HOPPE-SEYLER, 1996, 377 (03) :195-201
[33]   Zinc finger transcription factors mediate high constitutive platelet-derived growth factor-B expression in smooth muscle cells derived from aortae of newborn rats [J].
Rafty, LA ;
Khachigian, LM .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (10) :5758-5764
[34]  
ROBINSON D, 1995, GENE THER, V2, P269
[35]   PDGF B-CHAIN IN NEURONS OF THE CENTRAL-NERVOUS-SYSTEM, POSTERIOR PITUITARY, AND IN A TRANSGENIC MODEL [J].
SASAHARA, M ;
FRIES, JWU ;
RAINES, EW ;
GOWN, AM ;
WESTRUM, LE ;
FROSCH, MP ;
BONTHRON, DT ;
ROSS, R ;
COLLINS, T .
CELL, 1991, 64 (01) :217-227
[36]   A composite CMV-IE enhancer β-actin promoter is ubiquitously expressed in mouse cutaneous epithelium [J].
Sawicki, JA ;
Morris, RJ ;
Monks, B ;
Sakai, K ;
Miyazaki, J .
EXPERIMENTAL CELL RESEARCH, 1998, 244 (01) :367-369
[37]  
Veelken H, 1998, INT J MOL MED, V2, P423
[38]  
VIEWEG J, 1995, CANCER RES, V55, P2366
[39]   Enhanced suicide gene therapy by chimeric tumor-specific promoter based on HSF1 transcriptional regulation [J].
Wang, JH ;
Yao, MZ ;
Zhang, ZL ;
Gu, JF ;
Zhang, YH ;
Li, BH ;
Sun, LY ;
Liu, XY .
FEBS LETTERS, 2003, 546 (2-3) :315-320
[40]   CMV-β-actin promoter directs higher expression from an adeno-associated viral vector in the liver than the cytomegalovirus or elongation factor 1α promoter and results in therapeutic levels of human factor X in mice [J].
Xu, LF ;
Daly, T ;
Gao, CH ;
Flotte, TR ;
Song, SH ;
Byrne, BJ ;
Sands, MS ;
Ponder, KP .
HUMAN GENE THERAPY, 2001, 12 (05) :563-573