Ferrari Stefano, Pellegrini Graziella, Mavilio Fulvio, De Luca Michele
Epithelial Stem Cell Research Centre, Veneto Eye Bank Foundation, 30122 Venice, Italy.
Clin Dermatol. 2005 Jul-Aug;23(4):430-6. doi: 10.1016/j.clindermatol.2004.07.017.
Human epidermis consists of a stratified epithelium mainly composed of keratinocytes and relies on a stem cell compartment to undergo constant regeneration. Genetic mutations affecting the capacity of basal keratinocytes to adhere firmly to the epidermal basement membrane lead to severe, and very often lethal, blistering disorders known as epidermolysis bullosa. Gene therapy represents a promising potential treatment for these devastating inherited disorders. Human epidermal stem cells can be cultivated ex vivo and stably transduced with integrating gene transfer vectors, allowing genetic and, more important, phenotypic correction of the adhesion properties of keratinocytes. Here we will review some of the issues that need to be addressed to make gene therapy a realistic treatment for these disorders, such as (1) which cells should be targeted, (2) which approach (in vivo or ex vivo) should be chosen, and (3) which gene transfer vector (retrovirus, lentivirus, or integrating nonviral strategies) should be used for stable gene correction. In the last 10 years, many reports have shown that gene transfer approaches to target epidermal stem cells are feasible and able to restore the adhesion properties of primary keratinocytes from patients with epidermolysis bullosa. In addition, tremendous progress has been achieved in culturing epidermal stem cells and generating sheets of stratified epithelium for permanent coverage of full-thickness burns. Gene modification of stem cells in combination with advanced tissue-engineering techniques could therefore represent a realistic option for patients with epidermolysis bullosa.
人类表皮由主要由角质形成细胞组成的复层上皮构成，并依赖干细胞区室进行持续更新。影响基底角质形成细胞牢固黏附于表皮基底膜能力的基因突变会导致严重且常常致命的水疱性疾病，即大疱性表皮松解症。基因治疗是这些毁灭性遗传性疾病一种有前景的潜在治疗方法。人类表皮干细胞可以在体外培养，并用整合型基因转移载体进行稳定转导，从而实现角质形成细胞黏附特性的基因以及更重要的表型纠正。在此，我们将综述一些为使基因治疗成为这些疾病切实可行的治疗方法而需要解决的问题，例如：（1）应靶向哪些细胞；（2）应选择哪种方法（体内或体外）；以及（3）应使用哪种基因转移载体（逆转录病毒、慢病毒或整合型非病毒策略）进行稳定的基因纠正。在过去十年中，许多报告表明，靶向表皮干细胞的基因转移方法是可行的，并且能够恢复大疱性表皮松解症患者原代角质形成细胞的黏附特性。此外，在培养表皮干细胞以及生成用于永久覆盖全层烧伤的复层上皮片方面已经取得了巨大进展。因此，干细胞的基因修饰与先进的组织工程技术相结合可能是大疱性表皮松解症患者的一个切实可行的选择。
