Spirito Flavia, Capt Annabelle, Del Rio Marcela, Larcher Fernando, Guaguere Eric, Danos Olivier, Meneguzzi Guerrino
INSERM U634, Faculty of Medicine, Nice, France.
Biochem Biophys Res Commun. 2006 Jan 20;339(3):769-78. doi: 10.1016/j.bbrc.2005.10.216. Epub 2005 Nov 22.
Gene transfer represents the unique therapeutic issue for a number of inherited skin disorders including junctional epidermolysis bullosa (JEB), an untreatable genodermatose caused by mutations in the adhesion ligand laminin 5 (alpha3beta3gamma2) that is secreted in the extracellular matrix by the epidermal basal keratinocytes. Because gene therapy protocols require validation in animal models, we have phenotypically reverted by oncoretroviral transfer of the curative gene the keratinocytes isolated from dogs with a spontaneous form of JEB associated with a genetic mutation in the alpha3 chain of laminin 5. We show that the transduced dog JEB keratinocytes: (1) display a sustained secretion of laminin 5 in the extracellular matrix; (2) recover the adhesion, proliferation, and clonogenic capacity of wild-type keratinocytes; (3) generate fully differentiated stratified epithelia that after grafting on immunocompromised mice produce phenotypically normal skin and sustain permanent expression of the transgene. We validate an animal model that appears particularly suitable to demonstrate feasibility, efficacy, and safety of genetic therapeutic strategies for cutaneous disorders before undertaking human clinical trials.
基因转移是许多遗传性皮肤病面临的独特治疗问题，包括交界性大疱性表皮松解症（JEB），这是一种无法治愈的遗传性皮肤病，由粘附配体层粘连蛋白5（α3β3γ2）的突变引起，该蛋白由表皮基底角质形成细胞分泌到细胞外基质中。由于基因治疗方案需要在动物模型中进行验证，我们通过逆转录病毒转移治疗基因，使从患有与层粘连蛋白5α3链基因突变相关的自发性JEB的狗身上分离出的角质形成细胞的表型得到了恢复。我们发现，转导的犬JEB角质形成细胞：（1）在细胞外基质中持续分泌层粘连蛋白5；（2）恢复了野生型角质形成细胞的粘附、增殖和克隆形成能力；（3）产生完全分化的分层上皮，移植到免疫受损小鼠身上后可产生表型正常的皮肤，并维持转基因的永久表达。在进行人体临床试验之前，我们验证了一种动物模型，该模型似乎特别适合证明针对皮肤疾病的基因治疗策略的可行性、有效性和安全性。
