Our journey to successful gene therapy for hemophilia B.
Our journey to successful gene therapy for hemophilia B.
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我们成功治疗 B 型血友病基因疗法的历程。
DOI：
10.1089/hum.2014.2540
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发表时间：
2014
影响因子：
4.2
通讯作者：
Davidoff,AndrewM
中科院分区：
文献类型：
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作者：
Nathwani,AmitC;Nienhuis,ArthurW;Davidoff,AndrewM
1997年，我（ACN）在美国田纳西州孟菲斯市圣裘德儿童研究医院的Arthur Nienhuis博士的指导下，获得了为期3年的英国威康信托高级培训奖学金，研究使用腺相关病毒（aav）治疗镰状细胞病的基因治疗方法。在6个月内，我们意识到在病毒来源的载体中具有最佳安全性的AAV载体可能并不理想地适合镰状细胞病的基因治疗，因为它们在基因转移到人造血干细胞（hsc）后无法维持稳定的转基因表达。在基因转移后，AAV基因组主要以个体形式维持，并在造血干细胞分裂时迅速被抛弃（Nathwani et al., 2000）。为了寻找灵感，我回忆起我的博士主考人乔治·布朗利（George Brownlee）教授是如何分离出IX因子基因的，他建议我研究b型血友病的基因疗法。在尼恩休斯博士的支持下，我改变了思路，尽管那时其他研究小组已经抢先一步。特别是，该领域的先驱Katharine High博士当时正准备在血友病B患者中进行一项临床试验，该试验需要肌肉注射编码人类FIX的AAV载体。我们最初的研究重点是对小鼠模型中以肌肉、静脉和肝脏（产生因子IX的部位）为靶点的AAV递送方式的安全性和有效性进行正面比较。我向圣犹达大学外科学系新招募的教员Davidoff博士寻求帮助，以完成将AAV注入小鼠门静脉的艰巨任务。这是一段长期合作和友谊的开始。在进行这些实验的过程中，大卫杜夫博士和我开始互相交换意见，在他完全意识到之前，他已经完全参与了血友病基因治疗项目。我们发现，对于相同剂量的载体，与肌肉注射相比，全身或门静脉输送AAV后的表达明显更高。更令人担忧的事实是
In 1997, I (ACN) secured a 3-year Wellcome Trust Advanced Training Fellowship in the United Kingdom to pursue a gene therapy approach for sickle cell disease, using adeno-associated viruses (AAVs) under the supervision of Dr. Arthur Nienhuis at St. Jude Children’s Research Hospital, Memphis, Tennessee. Within 6 months, we realized that AAV vectors, which have the best safety profile among vectors of viral origin, may not be ideally suited for gene therapy of sickle cell disease because of their inability to maintain stable transgene expression following gene transfer into human hematopoietic stem cells (HSCs). The AAV genome is maintained predominantly in an episomal format following gene transfer and is rapidly jettisoned as HSCs undergo division (Nathwani et al., 2000). Searching for inspiration, I recalled how my Ph. D. examiner, Prof. George Brownlee, who had isolated the gene for factor IX, suggested that I work on gene therapy for hemophilia B. With Dr. Nienhuis’s support I, therefore, switched tack, though by then other groups had already stolen a march. In particular, Dr. Katharine High, a pioneer in this field, was at this time preparing to conduct a clinical trial in hemophilia B patients entailing intramuscular administration of AAV vectors encoding human FIX.Our initial efforts were focused on a head-to-head comparison of the safety and efficacy of the intramuscular, intravenous, and liver (the site of factor IX production)-targeted modes of AAV delivery in murine models. I sought help from Dr. Davidoff, a newly recruited faculty member in the Department of Surgery at St. Jude, for the challenging task of AAV administration into the portal vein of mice. This was the start of a long collaboration and friendship. In the course of conducting these experiments, Dr. Davidoff and I started bouncing ideas off each other, and before he fully realized, he had become integrally involved in the hemophilia gene therapy project. We discovered that, for the same dose of vector, expression was significantly higher following systemic or portal vein delivery of AAV when compared with intramuscular injections. More concerning was the fact that