Ex-vivo Gene Therapy Restores LEKTI Activity and Corrects the Architecture of Netherton Syndrome-derived Skin Grafts
Ex-vivo Gene Therapy Restores LEKTI Activity and Corrects the Architecture of Netherton Syndrome-derived Skin Grafts
复制标题
DOI：
10.1038/mt.2010.201
复制
发表时间：
2011-02-01
影响因子：
12.4
通讯作者：
Qasim, Waseem
中科院分区：
文献类型：
--
作者：
Di, Wei-Li;Larcher, Fernado;Qasim, Waseem
Netherton 综合征 (NS) 是一种使人衰弱的先天性皮肤病，由编码淋巴上皮 Kazal 型相关抑制剂 (LEKTI) 的 SPINK5 基因突变引起。其特点是角化缺陷、反复感染和高钠性脱水，出生后第一年死亡率约为 10%。目前，NS 没有治愈方法。我们开发了一种基于 HIV-1 的自失活慢病毒载体，可在角质形成细胞中表达 SPINK5，作为 NS 离体基因治疗策略的一部分。在 NS 角质形成细胞中实现了高转导效率，并且在先前缺陷的细胞中证实了 LEKTI 表达的重建。这些基因校正的角质形成细胞在体外器官培养（OTC）系统和体内小鼠/人类皮肤移植模型中进行了进一步测试。结果显示 OTC 和再生皮肤移植物的表皮结构均得到纠正。重要的是，校正后的皮肤移植结果表明，即使在有限数量的细胞中恢复了可检测的 LEKTI 表达，这些小群体周围的旁观者也会受益。由于 LEKTI 是一种分泌蛋白，因此转基因移植物不仅可以提供直接的局部保护屏障，而且还可以作为分泌型 LEKTI 的来源，在离体基因治疗后提供普遍的益处。
Netherton syndrome (NS) is a debilitating congenital skin disorder caused by mutations in the SPINK5 gene encoding the lymphoepithelial Kazal-type-related inhibitor (LEKTI). It is characterized by defective keratinization, recurrent infections, and hypernatraemic dehydration with a mortality rate of about 10% in the first year of life. Currently, there are no curative treatments for NS. We have developed a HIV-1 based, self-inactivating lentiviral vector to express SPINK5 in keratinocytes as part of an ex-vivo gene therapy strategy for NS. High transduction efficiency was achieved in NS keratinocytes and reconstitution of LEKTI expression was confirmed in previously deficient cells. These genetically corrected keratinocytes were further tested in an in vitro organotypic culture (OTC) system and in vivo mouse/human skin engraftment model. Results showed correction of epidermal architecture in both OTCs and regenerated skin grafts. Importantly, the results from corrected skin grafts indicated that even where detectable LEKTI expression was restored to a limited numbers of cells, a wider bystander benefit occurred around these small populations. As LEKTI is a secreted protein, the genetically modified graft may provide not only an immediate local protective barrier, but also act as a source of secreted LEKTI providing a generalized benefit following ex-vivo gene therapy.