Non-viral chimeric antigen receptor (CAR) T cells going viral.
Non-viral chimeric antigen receptor (CAR) T cells going viral.
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DOI：
10.1016/j.iotech.2023.100375
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发表时间：
2023-06
期刊：
影响因子：
--
通讯作者：
Feldman, S A
中科院分区：
文献类型：
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作者：
Balke-Want, H;Keerthi, V;Cadinanos-Garai, A;Fowler, C;Gkitsas, N;Brown, A K;Tunuguntla, R;Abou-El-Enein, M;Feldman, S A
嵌合抗原受体（CAR）T细胞疗法在B细胞恶性肿瘤的治疗方面取得了重大进展，但其在治疗实体瘤中的应用仍然面临着重大挑战。特别是，广泛使用病毒载体将 CAR 转基因传递到 T 细胞中存在局限性，包括高成本和监管限制，这阻碍了新颖的基因工程概念转化为临床应用。非病毒方法，例如转座子/转座酶和成簇规则间隔短回文重复序列 (CRISPR)/Cas 系统，为 CAR-T 细胞中稳定插入转基因提供了有前景的替代方案。这些方法有可能提高 CAR-T 细胞疗法的开发和交付的可及性和效率。然而，使用非病毒方法的主要挑战是其低敲入效率，这导致转基因表达水平低。在这篇综述中，我们讨论了 CAR-T 细胞非病毒方法的最新发展、非病毒 CAR-T 细胞临床级生产的制造要求，以及为正确表征基因组特征和评估潜在基因毒性而进行的质量控制调整。 CAR T 疗法病毒载体的高成本和监管障碍。非病毒基因递送可以克服病毒载体基因递送的局限性。低敲入效率仍然是非病毒 CAR-T 细胞生产中的一个挑战。重点关注非病毒载体设计和临床制造的进展。表征脱靶编辑和遗传毒性的分析方法。
Chimeric antigen receptor (CAR) T cell therapy has made significant strides in the treatment of B-cell malignancies, but its application in treating solid tumors still poses significant challenges. Particularly, the widespread use of viral vectors to deliver CAR transgenes into T cells comes with limitations, including high costs and regulatory restrictions, which hinder the translation of novel genetic engineering concepts into clinical applications. Non-viral methods, such as transposon/transposase and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas systems, offer promising alternatives for stable transgene insertion in CAR-T cells. These methods offer the potential to increase accessibility and efficiency in the development and delivery of CAR-T cell therapies. The main challenge in using non-viral methods, however, is their low knock-in efficiency, which leads to low transgene expression levels. In this review, we discuss recent developments in non-viral approaches for CAR-T cell production, the manufacturing requirements for clinical-grade production of non-viral CAR-T cells, and the adjustments needed in quality control for proper characterization of genomic features and evaluation of potential genotoxicity. High costs and regulatory hurdles associated with viral vectors for CAR T therapies. Non-viral gene delivery may overcome limitations of viral vector gene delivery. Low knock-in efficiency remains a challenge in non-viral CAR-T cell production. Focus on advancements in non-viral vector design and clinical manufacturing. Analytical methods to characterize off-target editing and genotoxicity.