Bacteria-free minicircle DNA system to generate integration-free CAR-T cells.
Bacteria-free minicircle DNA system to generate integration-free CAR-T cells.
复制标题
用于生成免整合 CAR-T 细胞的无菌小环 DNA 系统。
DOI：
10.1136/jmedgenet-2018-105405
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发表时间：
2019-01
影响因子：
4
通讯作者：
Wang H
中科院分区：
文献类型：
--
作者：
Cheng C;Tang N;Li J;Cao S;Zhang T;Wei X;Wang H
用慢病毒和逆转录病毒载体工程化的嵌合抗原受体T（CAR-T）细胞已成功应用于治疗患有B细胞恶性肿瘤的患者。然而，T细胞中的病毒整合具有诱变的潜在风险，并且病毒载体生产需要努力并且成本高。使用非整合附加型载体如小环载体来产生无整合的CAR-T细胞是一种有吸引力的选择。我们建立了一种新的方法，利用简单的分子生物学技术在几个小时内产生小环载体。由于不涉及细菌，我们将这些载体命名为无菌（BF）微环。与质粒相比，BF小环载体能够在人细胞系、干细胞和原代T细胞中实现更高的转基因表达并提高细胞活力。使用BF小环载体，我们产生了无整合的CAR-T细胞，其在体外和体内都有效地消除了癌细胞。BF小环载体将在基础研究以及CAR-T和基因治疗等临床应用中发挥重要作用。虽然小环载体的转基因表达持续时间明显短于插入慢病毒，但多轮BF小环CAR-T细胞输注可以有效消除癌细胞。另一方面，相对较短的CAR-T细胞持久性提供了避免严重副作用的机会，如细胞因子风暴或靶向脱靶毒性。
Chimeric antigen receptor T (CAR-T) cells engineered with lentiviral and retroviral vectors have been successfully applied to treat patients with B cell malignancy. However, viral integration in T cells has the potential risk of mutagenesis, and viral vector production demands effort and is costly. Using non-integrative episomal vector such as minicircle vector to generate integration-free CAR-T cells is an attractive option. We established a novel method to generate minicircle vector within a few hours using simple molecular biology techniques. Since no bacteria is involved, we named these vectors bacteria-free (BF) minicircle. In comparison with plasmids, BF minicircle vector enabled higher transgene expression and improved cell viability in human cell line, stem cells and primary T cells. Using BF minicircle vector, we generated integration-free CAR-T cells, which eliminated cancer cells efficiently both in vitro and in vivo. BF minicircle vector will be useful in basic research as well as in clinical applications such as CAR-T and gene therapy. Although the transgene expression of minicircle vector lasts apparently shorter than that of insertional lentivirus, multiple rounds of BF minicircle CAR-T cell infusion could eliminate cancer cells efficiently. On the other hand, a relatively shorter CAR-T cell persistence provides an opportunity to avoid serious side effects such as cytokine storm or on-target off-tumour toxicity.
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影响因子：
1.5
作者：
Gordillo, GM;Xia, DY;Orosz, CG
通讯作者：
Orosz, CG
影响因子：
2
作者：
Fischer, Alain
通讯作者：
Fischer, Alain
影响因子：
6.4
作者：
Nafissi, Nafiseh;Slavcev, Roderick
通讯作者：
Slavcev, Roderick
DOI：
10.1038/nrclinonc.2016.65
发表时间：
2016-06
期刊：
Nature reviews. Clinical oncology
影响因子：
--
作者：
Khalil DN;Smith EL;Brentjens RJ;Wolchok JD
通讯作者：
Wolchok JD
影响因子：
12.4
作者：
Chen, ZY;He, CY;Kay, MA
通讯作者：
Kay, MA