Comparison of cDNA and genomic forms of tyrosine hydroxylase gene therapy of the brain with Trojan horse liposomes.
Comparison of cDNA and genomic forms of tyrosine hydroxylase gene therapy of the brain with Trojan horse liposomes.
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特洛伊木马脂质体大脑酪氨酸羟化酶基因治疗的 cDNA 和基因组形式的比较。
DOI：
10.1002/jgm.1046
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发表时间：
2007
期刊：
影响因子：
--
通讯作者：
Pardridge,WilliamM
中科院分区：
文献类型：
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作者：
Xia,Chun-Fang;Chu,Chun;Li,Jianyi;Wang,Yuntao;Zhang,Yun;Boado,RubenJ;Pardridge,WilliamM
背景本研究探讨了染色体衍生形式的治疗基因是否可以在静脉给药后递送到脑中。大鼠酪氨酸羟化酶（TH）cDNA的脑表达进行了比较，脑表达的质粒DNA编码的18 kb的大鼠TH gene.MethodsTH基因的表达测定在细胞培养和在体内脑实验性帕金森病（PD）。总共设计了四种编码大鼠TH的真核表达质粒，其中TH表达盒的大小范围为1.5kb（在基因的cDNA形式的情况下）至17.5kb（在最大大小的基因组构建体的情况下）。TH表达质粒被递送到培养的细胞或大鼠脑体内与特洛伊木马脂质体（THL），其通过细胞膜受体将非病毒质粒DNA靶向细胞。结果TH基因在细胞培养物和体内的表达模式相似：TH基因的cDNA形式是快速作用的，作用持续时间短，TH基因的基因组形式是缓慢作用的，作用持续时间较长。纹状体TH酶活性在实验PD的最持续的更换产生的组合基因治疗的cDNA和TH基因的基因组形式的管理simultaneous.ConclusionsEukaryotic表达质粒编码的基因组形式的治疗基因，大至18 kb，可以成功地纳入THL和递送到脑静脉给药后。版权所有© 2007约翰威利父子有限公司。
BackgroundThe present study examines whether chromosomal derived forms of therapeutic genes can be delivered to brain following intravenous administration. The brain expression of a rat tyrosine hydroxylase (TH) cDNA is compared to the brain expression of a plasmid DNA encoding the 18 kb rat TH gene.MethodsTH gene expression is measured in cell culture andin vivoin brain in experimental Parkinson's disease (PD). A total of four eukaryotic expression plasmids encoding rat TH were engineered wherein the size of the TH expression cassette ranged from 1.5 kb, in the case of the cDNA form of the gene, to 17.5 kb, in the case of the largest size genomic construct. The TH expression plasmids were delivered to either cultured cells or to rat brainin vivowith Trojan horse liposomes (THLs), which target the non‐viral plasmid DNA to cells via cell membrane receptors.ResultsThe pattern of TH gene expression in cell culture andin vivowas similar: the cDNA form of the TH gene was fast‐acting with short duration of action, and the genomic form of the TH gene was slow‐acting with longer duration of action. The most sustained replacement of striatal TH enzyme activity in experimental PD was produced by combination gene therapy where both the cDNA and the genomic forms of the TH gene were administered simultaneously.ConclusionsEukaryotic expression plasmids encoding genomic forms of therapeutic genes, as large as 18 kb, can be successfully incorporated in THLs and delivered to brain following intravenous administration. Copyright © 2007 John Wiley & Sons, Ltd.