Toward epidermal stem cell-mediated ex vivo gene therapy of junctional epidermolysis bullosa
Toward epidermal stem cell-mediated ex vivo gene therapy of junctional epidermolysis bullosa
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DOI：
10.1089/104303400750035825
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发表时间：
2000-11-01
影响因子：
4.2
通讯作者：
De Luca, M
中科院分区：
文献类型：
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作者：
Dellambra, E;Pellegrini, G;De Luca, M
交界性大疱性表皮细胞瘤（JEB）是一组严重的遗传性皮肤病，由编码层粘连蛋白5或半桥粒其他成分的基因突变引起。由于人表皮是一种自我更新的组织，因此JEB的基因治疗需要将转基因稳定整合到表皮干细胞的基因组中。人表皮干细胞确实可以培养并用复制缺陷型逆转录病毒载体稳定转导，从而允许JEB角质形成细胞的粘附特性的完全表型校正。表皮干细胞产生适于永久覆盖大面积皮肤缺损的复层上皮的粘性片，并且遗传修饰的表皮片在移植到免疫缺陷动物上后保持转基因的长期表达。我们已经开发了一种临床方法，其允许在局部麻醉下将培养的表皮片移植到大的身体区域上，并且没有瘢痕结果。因此，（1）培养衬里上皮细胞的可能性，（2）非侵入性手术的可用性，允许移植大面积的皮肤，和（3）持续的转基因表达在体外和体内的表皮干细胞的演示，促使我们提出实施一个I/II期临床试验，旨在体外基因治疗选定的JEB患者。该试验的目的是在临床环境中验证离体程序，证明其整体安全性，并分析关键问题，如转基因植入物的长期存活，对转基因产品的免疫反应，以及转基因表达在治疗水平上的持续性。
Junctional epidermolysis bullosa (JEB) is a group of severe, inherited skin diseases caused by mutations in the genes encoding laminin 5 or other components of the hemidesmosome. Since human epidermis is a self-renewing tissue, gene therapy of JEB requires the stable integration of the transgene into the genome of the epidermal stem cell. Human epidermal stem cells can indeed be cultivated and stably transduced with replication-defective retroviral vectors, allowing full phenotypic correction of the adhesion properties of JEB keratinocytes, Epidermal stem cells generate cohesive sheets of stratified epithelium suitable for the permanent coverage of massive skin defects, and genetically modified epidermal sheets maintain long-term expression of the transgene after transplantation on immunodeficient animals, Moreover, we have developed a clinical procedure that allows transplantation of cultured epidermal sheets on large body areas under local anesthesia and without cicatricial outcomes. Thus, (1) the possibility of cultivating lining epithelia, (2) the availability of noninvasive surgical procedures that allow the grafting of large skin areas, and (3) the demonstration of sustained transgene expression in vitro and in vivo by epidermal stem cells, prompt us to propose the implementation of a phase I/II clinical trial aimed at the ex viva gene therapy of selected JEB patients. The aim of the trial is to validate the ex vivo procedure in a clinical setting, to prove its overall safety, and to analyze critical issues such as long-term survival of the genetically modified implant, immune response against the transgene product, and persistence of transgene expression at therapeutic levels.