Dupré Loïc, Marangoni Francesco, Scaramuzza Samantha, Trifari Sara, Hernández Raisa Jofra, Aiuti Alessandro, Naldini Luigi, Roncarolo Maria-Grazia
San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), 20132 Milan, Italy.
Hum Gene Ther. 2006 Mar;17(3):303-13. doi: 10.1089/hum.2006.17.303.
Wiskott-Aldrich syndrome (WAS) is a life-threatening X-linked primary immunodeficiency characterized by infections, hemorrhages, autoimmune disorders, and lymphomas. Transplantation of genetically corrected autologous hematopoietic stem cells (HSCs) could represent an alternative treatment to allogeneic HSC transplantation, the latter being often associated with severe complications. We used WAS-/- mice to test the efficacy of a gene therapy approach based on nonlethal irradiation followed by transplantation of WAS-/- HSCs transduced with lentiviral vectors encoding the WAS protein (WASP) from either the ubiquitous PGK promoter or the tissue- specific WAS promoter. The procedure resulted in significant levels of engraftment of WASP-expressing T cells, B cells, platelets, and myeloid cells. T cells harbored one or two vector copies and displayed partial to full correction of T cell receptor-driven interleukin-2 production and proliferation. In addition, polymerization of F-actin and localization of WASP at the site of the immunological synapse were restored. The treatment was well tolerated and no pathology was detected by systematic blood analysis and autopsy. The efficacy of WAS gene transfer into HSCs, using the WAS promoter-containing lentiviral vector, combined with nonlethal irradiation provides a strong rationale for the development of gene therapy for WAS patients.
威斯科特-奥尔德里奇综合征（WAS）是一种危及生命的X连锁原发性免疫缺陷病，其特征为感染、出血、自身免疫性疾病和淋巴瘤。基因校正的自体造血干细胞（HSC）移植可能是异基因HSC移植的一种替代治疗方法，而异基因HSC移植常常伴有严重并发症。我们使用WAS基因敲除小鼠来测试一种基因治疗方法的疗效，该方法基于非致死性照射，随后移植用携带来自遍在性磷酸甘油酸激酶（PGK）启动子或组织特异性WAS启动子且编码WAS蛋白（WASP）的慢病毒载体转导的WAS基因敲除HSC。该操作导致表达WASP的T细胞、B细胞、血小板和髓系细胞出现显著水平的植入。T细胞含有一或两个载体拷贝，并表现出T细胞受体驱动的白细胞介素-2产生和增殖的部分至完全校正。此外，F-肌动蛋白的聚合以及WASP在免疫突触部位的定位得以恢复。该治疗耐受性良好，系统血液分析和尸检未检测到病理学变化。使用含WAS启动子的慢病毒载体将WAS基因转移至HSC，并结合非致死性照射，为开发针对WAS患者的基因治疗提供了有力依据。
