In vivo hematopoietic stem cell gene therapy ameliorates murine thalassemia intermedia
In vivo hematopoietic stem cell gene therapy ameliorates murine thalassemia intermedia
复制标题
DOI：
10.1172/jci122836
复制
发表时间：
2019-02-01
影响因子：
15.9
通讯作者：
Lieber, Andre
中科院分区：
文献类型：
--
作者：
Wang, Hongjie;Georgakopoulou, Aphrodite;Lieber, Andre
目前的地中海贫血基因治疗方案需要收集造血干/祖细胞（HSPC），体外培养，慢病毒载体转导，并再次移植到骨髓清除患者。由于成本和技术复杂性，这种方案不太可能适用于发展中国家，而发展中国家对β地中海贫血疗法的需求最大。我们已经开发了一种简单的体内HSPC基因治疗方法，其涉及HSPC动员和整合HDAd 5/35++载体的静脉内注射。转导的HSPCs回到骨髓，在那里它们长期存在。用于地中海贫血的体内基因治疗的HDAd 5/35++载体具有通过人CD 46靶向原始HSPC的独特衣壳、相对安全的基于SB 100 X转座酶的整合机制、微LCR驱动的γ-珠蛋白基因和MGMT（P140 K）系统，其允许通过用低剂量O-6-苄基鸟嘌呤加双-氯乙基亚硝基脲短期治疗来增加治疗效果。我们在“健康”的人CD 46转基因小鼠和中间型地中海贫血小鼠模型中表明，我们的体内方法导致大多数循环红细胞中稳定的γ-珠蛋白表达。高标记频率保持在二级受体。在地中海贫血模型中，实现了接近完全的表型校正。治疗耐受性良好。这种具有成本效益和“便携式”的方法可以允许地中海贫血基因治疗的更广泛的临床应用。
Current thalassemia gene therapy protocols require the collection of hematopoietic stem/progenitor cells (HSPCs), in vitro culture, lentivirus vector transduction, and retransplantation into myeloablated patients. Because of cost and technical complexity, it is unlikely that such protocols will be applicable in developing countries, where the greatest demand for a beta-thalassemia therapy lies. We have developed a simple in vivo HSPC gene therapy approach that involves HSPC mobilization and an intravenous injection of integrating HDAd5/35++ vectors. Transduced HSPCs homed back to the bone marrow, where they persisted long-term. HDAd5/35++ vectors for in vivo gene therapy of thalassemia had a unique capsid that targeted primitive HSPCs through human CD46, a relatively safe SB100X transposase-based integration machinery, a micro-LCR-driven gamma-globin gene, and an MGMT(P140K) system that allowed for increasing the therapeutic effect by short-term treatment with low-dose O-6-benzylguanine plus bis-chloroethylnitrosourea. We showed in "healthy" human CD46-transgenic mice and in a mouse model of thalassemia intermedia that our in vivo approach resulted in stable gamma-globin expression in the majority of circulating red blood cells. The high marking frequency was maintained in secondary recipients. In the thalassemia model, a near-complete phenotypic correction was achieved. The treatment was well tolerated. This cost-efficient and "portable" approach could permit a broader clinical application of thalassemia gene therapy.