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

试点研究——单纯疱疹病毒用于 LESCH-NYHAN 综合征的基因治疗

• 批准号: 2374251
• 金额: --
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2374251
• 关键词: Lesch Nyhan syndrome; gene expression; gene therapy; genetically modified animals; herpes simplex virus 1; hypoxanthine phosphoribosyltransferase; laboratory mouse; latent virus infection; recombinant DNA; tissue /cell culture; transfection /expression vector; virus replication

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.

许多遗传性疾病影响神经系统。的长期目标