Department of Surgery, University Medical Center Regensburg, Regensburg, Germany.
J Gastroenterol Hepatol. 2010 May;25(5):1002-8. doi: 10.1111/j.1440-1746.2009.06156.x.
Despite pharmaceutical treatment with NTBC (2-2-nitro-4-fluoromethylbenzoyl-1,3-cyclohexanedione), a high incidence of liver malignancies occur in humans and mice suffering from hereditary tyrosinemia type 1 (HT1) caused by mutation of the fumarylacetoacetate hydrolase (fah) gene.
To evaluate the efficacy of a definitive treatment for HT1, we transfected fah knockout mice with naked plasmid DNA using high volume tail-vein injection. This approach was chosen to reduce the occurrence of insertional mutagenesis that is frequently observed when using other (retro-)viral vectors. To prolong gene expression, the fah gene was cloned between adeno-associated virus (AAV)-specific inverted terminal repeats (ITRs).
All animals treated with high volume plasmid DNA injections could be successfully weaned off NTBC and survived in the long term without any further pharmacological support. Up to 50% fah positive hepatocytes were detected in livers of naked plasmid DNA-treated animals and serum liver function tests approximated those of wild-type controls.
Naked plasmid DNA transfection offers a promising alternative treatment for HT1. Minimizing side-effects makes this approach especially appealing.
尽管使用 NTBC（2-2-硝基-4-氟甲基苯甲酰-1,3-环己二酮）进行药物治疗，但遗传性酪氨酸血症 1 型（HT1）患者仍会发生很高的肝脏恶性肿瘤发病率，这种疾病是由 fumarylacetoacetate hydrolase（fah）基因突变引起的。
为了评估针对 HT1 的根治性治疗方法的疗效，我们使用大体积尾静脉注射的方法，将 fah 基因敲除小鼠转染裸质粒 DNA。之所以选择这种方法，是为了减少在使用其他（逆转录）病毒载体时经常观察到的插入突变。为了延长基因表达，fah 基因被克隆到腺相关病毒（AAV）特异性的反向末端重复序列（ITR）之间。
所有接受大体积质粒 DNA 注射治疗的动物均成功地摆脱了 NTBC 的治疗，并在没有任何进一步药物支持的情况下长期存活。裸质粒 DNA 处理的动物肝脏中可检测到高达 50%的 fah 阳性肝细胞，血清肝功能检测接近野生型对照。
裸质粒 DNA 转染为 HT1 提供了一种有前途的替代治疗方法。减少副作用使这种方法特别有吸引力。
