PILOT STUDY--HERPES SIMPLEX VIRUS FOR GENE THERAPY FOR LESCH-NYHAN SYNDROME

许多遗传性疾病会影响神经系统。的长期目标是 这项拨款计划是为了开发无毒的1型单纯疱疹病毒。 (HSV-1)株作为神经系统疾病的基因治疗载体。 HSV-1感染有两个特征将被利用 为了实现这一目标：HSV-1在中国建立终身潜伏感染 在潜伏感染期间，只有一个病毒启动子是 激活。该启动子指导HSV-1潜伏期的合成 相关转录本(LAT)，在生命周期内表达 潜伏感染的个体。为了证明HSV-1的可行性-- 人次黄嘌呤-鸟嘌呤的基因治疗 磷酸核糖基转移酶(HPRT)基因将置于 HSV-1 LAT基因启动子。HPRT的完全缺陷导致Lesch- 尼汉综合征，一种严重且无法治愈的神经系统疾病。自.以来 HPRT水平非常低的个体不会受到神经系统疾病的影响 功能障碍，甚至部分替代神经系统中的HPRT可能是 有治疗作用。HPRT阴性转基因小鼠可作为动物使用 评估基因转移效果的Lesch-Nyhan综合征模型 技巧。目前有几种无毒力的HSV-1毒株可供选择，两者 复制能力强和复制能力弱。这些病毒不会产生任何 小鼠的疾病，即使在大脑内接种时也是如此(K.C.)。发展，发展 对于HSV-1向神经系统的基因输送，我们将i)研究几个 通过神经系统传播的无毒HSV-1毒株， 急性感染期间的细胞病理学、病毒RNA和蛋白质表达， 以及LAT表达细胞的类型、数量和分布 潜伏期，ii)确定POLYA+/调节序列要求 HSV-1潜伏期启动子在MOST中的表达水平 有希望的无毒菌株(S和III)，组织调查 人HPRT基因及其蛋白在正常小鼠和HPRT小鼠体内的分布 HSV-1/hprt重组体感染阴性转基因小鼠。

Many genetic diseases affect the nervous system. The long term goal of this grant proposal is to develop avirulent herpes simplex virus type 1 (HSV-1) strains as gene therapy vectors for nervous system disorders. There are two characteristics of HSV-1 infections that will be exploited to achieve this goal: HSV-1 establishes life-long latent infections in neurons, and during latent infection only a single viral promoter is active. This promoter directs the synthesis of the HSV-1 latency associated transcripts (LATs), which are expressed for the lifetime of the latently infected individual. To demonstrate the feasibility of HSV-1- mediated gene therapy, a cDNA for the human hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene will be placed under the control of the HSV-1 LAT gene promoter. A complete deficiency of HPRT causes Lesch- Nyhan syndrome, a severe and untreatable neurological disease. Since individuals with very low HPRT levels are spared from neurological dysfunction, even partial replacement of HPRT in the nervous system may be therapeutic. HPRT-negative transgenic mice are available as an animal model of Lesch-Nyhan syndrome to assess the effectiveness of gene transfer techniques. There are several avirulent HSV-1 strains available, both replication competent and incompetent. These viruses do not produce any disease in mice, even when inoculated intracerebrally (k.c.). To develop HSV-1 for gene delivery to the nervous system, we will i) study several avirulent HSV-1 strains for spread through the nervous system, cytopathology, viral RNA and protein expression during acute infection, and the type, number and distribution of LAT expressing cells during latency, ii)determine the polyA+/regulatory sequence requirements for high levels of HSV-1 latency promoter directed cDNA expression in the most promising avirulent strains(s), and iii) investigate the tissue distribution of human HPRT mRNA and proteins in normal mice and in HPRT- negative transgenic mice infected with the HSV-1/HPRT recombinants.