Semi-automated closed system manufacturing of lentivirus gene-modified haematopoietic stem cells for gene therapy
Semi-automated closed system manufacturing of lentivirus gene-modified haematopoietic stem cells for gene therapy
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DOI：
10.1038/ncomms13173
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发表时间：
2016-10-20
影响因子：
16.6
通讯作者：
Kiem, Hans-Peter
中科院分区：
文献类型：
--
作者：
Adair, Jennifer E.;Waters, Timothy;Kiem, Hans-Peter
造血干细胞（HSC）基因治疗已被证明具有治疗许多疾病的潜力。然而，现有技术需要在专用的良好生产规范设施中进行复杂的离体基因转移，限制了可用性。一个自动化的过程将提高HSC基因治疗的可用性和标准化生产。在这里，我们开发了一种新的程序，半自动细胞分离和培养设备，以允许完整的台式生产的基因修饰的CD34（+）血细胞产品的移植。这些细胞产品符合动员白细胞去除术和骨髓的现行生产质量标准，并在免疫功能低下的小鼠中重建人造血。重要的是，非人灵长类动物自体基因修饰的CD34（+）细胞产物能够在超过1年的随访中进行稳定的多克隆多谱系重建。这些数据证明了HSC基因治疗的床旁递送的概念证明。鉴于基因治疗的靶点疾病很多，这种方法在全球范围内治疗患者的潜力巨大。
Haematopoietic stem cell (HSC) gene therapy has demonstrated potential to treat many diseases. However, current state of the art requires sophisticated ex vivo gene transfer in a dedicated Good Manufacturing Practices facility, limiting availability. An automated process would improve the availability and standardized manufacture of HSC gene therapy. Here, we develop a novel program for semi-automated cell isolation and culture equipment to permit complete benchtop generation of gene-modified CD34(+) blood cell products for transplantation. These cell products meet current manufacturing quality standards for both mobilized leukapheresis and bone marrow, and reconstitute human haematopoiesis in immunocompromised mice. Importantly, nonhuman primate autologous gene-modified CD34(+) cell products are capable of stable, polyclonal multilineage reconstitution with follow-up of more than 1 year. These data demonstrate proof of concept for point-of-care delivery of HSC gene therapy. Given the many target diseases for gene therapy, there is enormous potential for this approach to treat patients on a global scale.