Longterm gene therapy of xeroderma pigmentosum group A mice using HVJ-liposomes.

使用 HVJ 脂质体对着色性干皮病 A 组小鼠进行长期基因治疗。

基本信息

批准号：
14370258
负责人：
NISHIGORI Chikako
金额：
$ 5.31万
依托单位：
Kobe University (2003)Kyoto University (2002)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

Xeroderma pigmentosum (XP) is an autosomalrecessibely inheritated disorder in which nucleotide excision repair is deficient, thus the patients with XP cause skin cancer of sun-exposed area in high frequency. Among XP complementation groups, XP complementation group A (XPA) reveals the severest clinical phenotype with neurological abnormalities and XPA is the most common type among Japanese XPA patients. The aim of this project is to challenge the gene therapy of XPA model mice in vivo with treating human XPA gene in the form of HVJ-liposomes, since one of our research goal is to establish the gene therapy for XPA patients. Two plasmids containing human XPA gene, pCAGGS-XPA or pcHA-XPA, which contains hemagultinin gene as a tag, was transfected and the repair ability was measured by unscheduled DNA syntshesis at post 24 or 48 hr UVC irradiation. Both recovered the repair efficiency in XPA mice cells. In in vivo study HVJ (Hemagluutinating Virus of Japan)-liposomes was used for introducing the plasmid containing human XPA gene into mice skin. HVJ-liposomes-XPA was injected intradermaly 24 hr before 4 kJ/m^2 of UVB irradiation or topically painted 48 hr before UVB irradiation. Unscheduled DNA synthesis of epidermal nuclei in mice injected XPA-HVJ liposmes was higher than that of control liposomes. Next we irradiated the mice with 500 J/m^2 of UVB 1-2 times per week for 3 months with or without treating HVJ-XPA-liposmes 24 or 48 hr prior to UVB irradiation. Both pcHA-XPA and pCAGGS-XPA encapsulated in HVJ-liposmes reduced the size, the number of the UV-induced tumors and elongate the survival duration. However, both treatment did not completely inhibit the tumor formation. The histology of the tumor was squamous cell carcinoma. In conclusion, the treatment of HVJ-XPA liposomes has effect on reducing the UV-induced tumors in XPA model mice.

心虫色素（XP）是一种常染色体遗传性疾病，其中核苷酸切除修复不足，因此XP患者在高频中引起阳光暴露区域的皮肤癌。在XP互补组中，XP互补组A（XPA）揭示了具有神经系统异常的最严重的临床表型，而XPA是日本XPA患者中最常见的类型。该项目的目的是挑战XPA模型小鼠在体内的基因治疗，并以HVJ-脂质体的形式治疗人XPA基因，因为我们的研究目标之一是为XPA患者建立基因治疗。转染了两个含有人XPA基因的质粒，其中包含人XPA基因的PCAGGS-XPA或PCHA-XPA，其中包含hragultin蛋白基因作为TAG，并通过在24或48 hr UVC辐照后的未定义的DNA Syntshesses测量修复能力。两者都恢复了XPA小鼠细胞中的修复效率。在体内研究中，使用HVJ（日本的血液升高病毒） - 脂质体用于将含有人XPA基因的质粒引入小鼠皮肤中。在UVB辐射的4 kJ/m^2之前，将HVJ - 脂质体-XPA注射24小时内24小时，或在UVB辐照前局部涂漆48小时。在注射XPA-HVJ脂质的小鼠中表皮核的外科DNA合成高于对照脂质体的DNA。接下来，我们在UVB辐照之前用或不处理HVJ-XPA-脂质24或48小时的UVB每周1-2次的500 J/m^2次，持续3个月。封装在HVJ-liposmes中的PCHA-XPA和PCAGGS-XPA都减少了紫外线诱导的肿瘤的大小，并延长了存活持续时间。但是，两种治疗都没有完全抑制肿瘤的形成。肿瘤的组织学是鳞状细胞癌。总之，HVJ-XPA脂质体的治疗对减少XPA模型小鼠紫外线诱导的肿瘤的影响。