PDF(Pubmed)
Abstract：
Chimeric antigen receptor (CAR) T-cell therapy has proven a breakthrough in cancer treatment in the last decade, giving unprecedented results against hematological malignancies. All approved CAR T-cell products, as well as many being assessed in clinical trials, are generated using viral vectors to deploy the exogenous genetic material into T-cells. Viral vectors have a long-standing clinical history in gene delivery, and thus underwent iterations of optimization to improve their efficiency and safety. Nonetheless, their capacity to integrate semi-randomly into the host genome makes them potentially oncogenic via insertional mutagenesis and dysregulation of key cellular genes. Secondary cancers following CAR T-cell administration appear to be a rare adverse event. However several cases documented in the last few years put the spotlight on this issue, which might have been underestimated so far, given the relatively recent deployment of CAR T-cell therapies. Furthermore, the initial successes obtained in hematological malignancies have not yet been replicated in solid tumors. It is now clear that further enhancements are needed to allow CAR T-cells to increase long-term persistence, overcome exhaustion and cope with the immunosuppressive tumor microenvironment. To this aim, a variety of genomic engineering strategies are under evaluation, most relying on CRISPR/Cas9 or other gene editing technologies. These approaches are liable to introduce unintended, irreversible genomic alterations in the product cells. In the first part of this review, we will discuss the viral and non-viral approaches used for the generation of CAR T-cells, whereas in the second part we will focus on gene editing and non-gene editing T-cell engineering, with particular regard to advantages, limitations, and safety. Finally, we will critically analyze the different gene deployment and genomic engineering combinations, delineating strategies with a superior safety profile for the production of next-generation CAR T-cell.
摘要：
嵌合抗原受体（CAR）T细胞疗法在过去十年中被证明是癌症治疗的突破。对血液恶性肿瘤产生前所未有的效果。所有批准的CAR-T细胞产品，以及许多正在临床试验中评估的人，使用病毒载体将外源遗传物质部署到T细胞中产生。病毒载体在基因传递中具有长期的临床历史，因此进行了迭代优化，以提高其效率和安全性。尽管如此，它们半随机整合到宿主基因组中的能力使它们可能通过插入诱变和关键细胞基因的失调而致癌。CART细胞施用后的继发性癌症似乎是罕见的不良事件。然而，过去几年记录的几起案件引起了人们对这个问题的关注，到目前为止可能被低估了，考虑到相对较新的CART细胞疗法的部署。此外，在血液恶性肿瘤中获得的最初成功尚未在实体瘤中复制。现在很明显，需要进一步的增强才能使CAR-T细胞增加长期持久性，克服疲惫，应对免疫抑制肿瘤微环境。为了这个目标,各种基因组工程策略正在评估中，最依赖CRISPR/Cas9或其他基因编辑技术。这些方法很容易引入意想不到的，产物细胞中不可逆的基因组改变。在本文的第一部分,我们将讨论用于产生CAR-T细胞的病毒和非病毒方法，而在第二部分，我们将专注于基因编辑和非基因编辑T细胞工程，特别是在优势方面，局限性,和安全。最后,我们将批判性地分析不同的基因部署和基因组工程组合，描述具有卓越安全性的下一代CAR-T细胞生产策略。
