细胞和基因治疗产品整合位点研究进展

doi:10.19803/j.1672-8629.20250151

摘要/Abstract

摘要： 目的系统阐述逆转录病毒与重组腺相关病毒载体的插入整合机制,总结整合毒性评价方法和整合位点分析技术方法,加强对细胞和基因治疗（CGT）产品安全性的关注。方法检索中国知网、PubMed 等数据库,从病毒载体整合位点（Integration Site, IS）的毒性机制和研究方法两个方面,整理和分析不同类型载体的整合位点研究内容。结果逆转录病毒载体具有高频随机整合特点,采用体外细胞永生化法（IVIM）、改良IVIM或转录物检测方法评估整合毒性。重组腺相关病毒载体具有低频定向整合特点,采用新生甲基丙二酸血症小鼠试验评估整合毒性,血液循环核酸是更合适的表征样本。且对比全基因组测序、连接介导PCR法、线性扩增介导PCR法、非限制性扩增PCR法、改良基因组测序PCR法、ITR-seq和AAV-seq等整合位点分析技术方法的优势与局限性。结论病毒载体是生产CGT产品的主要转基因工具,开展整合位点研究对确保产品安全性和预防临床药品不良反应至关重要。

关键词: 细胞和基因治疗, 病毒载体, 逆转录病毒, 慢病毒, 重组腺相关病毒, 整合位点, 评价模型, 二代测序

Abstract: Objective To elaborate the insertional integration mechanisms of retroviral and recombinant adeno-associated viral (rAAV) vectors, and review assessment Methods of integrated toxicity and analytic approaches to integration sites in order to call attention to the safety of cellular and gene therapy (CGT) products. Methods CNKI, PubMed and other databases were searched for literature related to integration sites of different vector types in general and toxicity mechanisms and investigational Methods in particular. Results Retroviral vectors were characterized by high-frequency random integration. The integrated toxicity was assessed using in vitro immortalization assay (IVIM), modified IVIM, or the transcript detection method. rAAV vectors were capable of low-frequency targeted integration, and the integrated toxicity was assessed using a neonatal mouse model of methylmalonic acidemia, where circulating cell-free nucleic acid was a more suitable characterization specimen. Besides, advantages and limitations of analytic techniques for integration sites, including WGS, LM-PCR, LAM-PCR, nr LAM-PCR, MGS-PCR, ITR-seq and AAV-seq, were compared. Conclusion Viral vectors are the primary gene delivery tool for the making of CGT products. The analysis of integration sites of vectors is crucial for ensuring product safety and preventing adverse drug reactions.

Key words: Cellular and Gene Therapy, Viral Vectors, Retrovirus, Lentivirus, Recombinant Adeno-Associated Virus, Integration Site, Assessment Model, Next-Generation Sequencing

中图分类号:

R954.5

綦玉洁, 王欣, 张嘉慧, 马恩龙, 耿兴超. 细胞和基因治疗产品整合位点研究进展[J]. 中国药物警戒, 2025, 22(7): 721-727.

QI Yujie, WANG Xin, ZHANG Jiahui, MA Enlong, GENG Xingchao. Research Progress in Integration Sites for Cellular and Gene Therapy Products[J]. Chinese Journal of Pharmacovigilance, 2025, 22(7): 721-727.

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