Development of new gene therapy for metabolic disorder of the liver -in vivo gene transfection using Particle Delivery System-

我们研究了利用基因枪进行体内基因转移，以前称为遗传性肝病的颗粒递送系统。为了建立这种方法，我们测试了4种基因枪(Geneblaster 1221、Idera gie-III、锤式和手持式)在活体大鼠的培养细胞和肝脏中的表达。将包被CMVβ-半乳糖苷酶的金粒子轰击入体外和体内。对两种体系中的半乳糖苷酶活性进行了估算。将包被CMVA-18(UDP-葡萄糖醛酸基转移酶)的Au颗粒多次注入缺乏胆红素UDP-葡萄糖醛酸基转移酶突变体的Gunn大鼠的肝脏中，使用新开发的手持式基因递送系统获得了最好的结果。在250psi的氦气压力下，将硝酸铵颗粒导入大鼠肝脏，在有限的肝脏表面区域(8×8 mm，深度2 mm)表达了β-半乳糖苷基因(1.2mU/µg蛋白质)。但血清胆红素水平并未恢复至正常大鼠水平。干细胞的体外基因转导被认为是治疗全身性代谢紊乱的理想方法，而基因枪在细胞内的转导具有快速、简便、通用性强等优点。将多种DNA混合在一起，可能为实体瘤的治疗提供一种新的有效方法。

We studied the in vivo gene transfection using gene gun, formerly called particle delivery system for congenially hepatic disorder disease. To develop this method, we tested 4 kinds of gene guns (Geneblaster 1221, Idera Gie-III,Hammer type, and Hand-held type) in culture cells and liver of living rats. Gold particles coated with CMV beta-galactosidase were bombarded into ex vivo and in vivo. The galactosidase activity was estimated in both systems. Au particles coated with CMV A-18 (UDP-glucuronosy transferase) were also done several times into the liver of Gunn rats which have a mutant lacking bilirubin UDP-glucuronosy transferase.The best result was obtained by using a newly developed hand-held gene delivery system. The beta-galactosidose gene introduced into the rat liver with the An particle by He gas pressure of 250 psi, was expressed (1.2 mu units/mu g protein) in a limited area of the liver surface (8*8mm, depth 2mm). However, the serum level of bilirubine was not returned to that of the normal rats. Ex vivo gene transfection to the stem cells has been considered to be satiable for systemic metabolic disorder, but the in situ intracellular delivery with gene gun was fast, simple and versatile. A new effective method might be developed for solid cancer by mixing several kinds of DNA.