Palella T D, Hidaka Y, Silverman L J, Levine M, Glorioso J, Kelley W N
Department of Internal Medicine, University of Michigan Medical School, Ann Arbor 48109.
Gene. 1989 Aug 1;80(1):137-44. doi: 10.1016/0378-1119(89)90258-8.
Complete deficiency of the purine salvage enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) results in a devastating neurological disease, the Lesch-Nyhan syndrome. This disorder has been identified as a candidate for initial attempts at somatic cell gene therapy. We have previously reported the construction of a recombinant herpes simplex virus type 1 (HSV-1) vector containing human hprt cDNA sequences under the regulatory control of the viral thymidine kinase gene (tk) [Palella et al., Mol. Cell. Biol. 8 (1988) 457-460]. Infection of HPRT- cultured rat neuronal cells with these vectors resulted in transient expression of human hprt. In this paper, we report the expression of human hprt mRNA transcripts in the brains of mice infected in vivo with this vector by direct intracranial inoculation. Human hprt transcripts were distinguished from endogenous mouse transcripts by RNase A mapping using riboprobes transcribed from human hprt cDNA. These initial studies demonstrate the transfer and transcription of a human gene in brain cells by direct in vivo infection with recombinant HSV-1 vectors.
嘌呤补救酶次黄嘌呤 - 鸟嘌呤磷酸核糖转移酶（HPRT）的完全缺乏会导致一种毁灭性的神经疾病——莱施 - 尼汉综合征。这种疾病已被确定为体细胞基因治疗初步尝试的候选对象。我们之前报道过构建一种重组单纯疱疹病毒1型（HSV - 1）载体，该载体在病毒胸苷激酶基因（tk）的调控下包含人hprt cDNA序列[帕莱拉等人，《分子与细胞生物学》8（1988）457 - 460]。用这些载体感染培养的HPRT大鼠神经元细胞会导致人hprt的瞬时表达。在本文中，我们报告了通过直接颅内接种，用该载体在体内感染小鼠后，人hprt mRNA转录本在小鼠脑中的表达情况。使用从人hprt cDNA转录的核糖探针通过核糖核酸酶A图谱分析，将人hprt转录本与内源性小鼠转录本区分开来。这些初步研究证明了通过重组HSV - 1载体直接体内感染，人类基因可在脑细胞中进行转移和转录。
