Engineering HSV-1 Vectors for Gene Therapy
Engineering HSV-1 Vectors for Gene Therapy
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DOI：
10.1007/978-1-4939-0428-0_5
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发表时间：
2014-01-01
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影响因子：
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通讯作者：
Glorioso, Joseph C.
中科院分区：
文献类型：
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作者：
Goins, William F.;Huang, Shaohua;Glorioso, Joseph C.
病毒载体已被用作各种临床前和临床基因治疗应用的基因转移载体，并且随着Glybera（alipogene tiparvovec）被批准作为第一种基因治疗产品作为标准医学治疗（Yla-Herttuala，Mol Ther 20：1831-1832，2013），基因治疗已经达到作为标准患者护理的一部分的状态。在活跃分裂的肿瘤细胞中特异性复制的有复制能力的单纯疱疹病毒（HSV）载体已用于多形性胶质母细胞瘤（一种致命形式的脑癌）和恶性黑色素瘤患者的I-III期人体试验。事实上，当与单独的标准GM-CSF治疗相比时，T-VEC（talimogene laherparepvec，以前称为OncoVex GM-CSF）在最近的III期试验中显示出功效（Andtbacka等人，J Clin Oncol 31：sLBA 9008，2013），并且可能很快成为用于标准患者护理的第二种FDA批准的基因治疗产品。除了有复制能力的溶瘤HSV载体如T-VEC之外，复制缺陷型HSV载体已经用于I-II期人体试验，并且已经被探索作为病症如疼痛、神经病和其他神经退行性病症的递送载体。在过去十年中对HSV载体开发的研究已经导致了重组载体的工程化，所述重组载体是完全复制缺陷型的、无毒的并且能够在神经元中长期转基因表达。本章描述了基于HSV-1复制缺陷型载体骨架构建重组基因组HSV载体的方法、纯化步骤以及用于细胞培养实验和临床前动物研究的小规模生产。
Virus vectors have been employed as gene transfer vehicles for various preclinical and clinical gene therapy applications, and with the approval of Glybera (alipogene tiparvovec) as the first gene therapy product as a standard medical treatment (Yla-Herttuala, Mol Ther 20: 1831-1832, 2013), gene therapy has reached the status of being a part of standard patient care. Replication-competent herpes simplex virus (HSV) vectors that replicate specifically in actively dividing tumor cells have been used in Phase I-III human trials in patients with glioblastoma multiforme, a fatal form of brain cancer, and in malignant melanoma. In fact, T-VEC (talimogene laherparepvec, formerly known as OncoVex GM-CSF) displayed efficacy in a recent Phase III trial when compared to standard GM-CSF treatment alone (Andtbacka et al. J Clin Oncol 31: sLBA9008, 2013) and may soon become the second FDA-approved gene therapy product used in standard patient care. In addition to the replication-competent oncolytic HSV vectors like T-VEC, replication-defective HSV vectors have been employed in Phase I-II human trials and have been explored as delivery vehicles for disorders such as pain, neuropathy, and other neurodegenerative conditions. Research during the last decade on the development of HSV vectors has resulted in the engineering of recombinant vectors that are totally replication defective, nontoxic, and capable of long-term transgene expression in neurons. This chapter describes methods for the construction of recombinant genomic HSV vectors based on the HSV-1 replication-defective vector backbones, steps in their purification, and their small-scale production for use in cell culture experiments as well as preclinical animal studies.