Retrieval of vector integration sites from cell-free DNA
Retrieval of vector integration sites from cell-free DNA
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DOI：
10.1038/s41591-021-01389-4
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发表时间：
2021-06-17
期刊：
影响因子：
82.9
通讯作者：
Montini, Eugenio
中科院分区：
文献类型：
--
作者：
Cesana, Daniela;Calabria, Andrea;Montini, Eugenio
一种称为液体活检整合位点测序的新方法可以监测接受基因治疗的患者实体组织中的转基因细胞。基因治疗（GT）作为一种治疗其他不治之症的方法，迅速引起了人们的新兴趣，几种GT产品已经上市，更多的产品进入了选定适应症的临床试验。基于载体整合的克隆跟踪技术能够监测接受GT的患者血液中工程细胞的命运，并评估这些程序的安全性和有效性。然而，由于可以测试的细胞数量有限，并且在不进行侵入性活检的情况下研究驻留在外周器官中的细胞是不切实际的，这种方法仅提供了遗传修饰细胞的克隆库和动态的部分快照，并且降低了作为安全读数的预测能力。在这项研究中，我们开发了液体活检整合位点测序，或LiBIS-seq，聚合酶链反应技术优化，定量检索载体整合位点从游离DNA释放到血流中的死亡细胞驻留在几个组织。这种方法使纵向监测体内肝定向GT和克隆跟踪接受造血干细胞GT的患者，提高我们的理解克隆组成和转基因细胞在实体组织中的营业额，并在传统的分析仅基于循环血细胞，使载体标记的克隆早期检测异常扩增外周组织。
A new approach called liquid biopsy integration site sequencing enables monitoring of genetically modified cells in solid tissues of patients receiving gene therapy.Gene therapy (GT) has rapidly attracted renewed interest as a treatment for otherwise incurable diseases, with several GT products already on the market and many more entering clinical testing for selected indications. Clonal tracking techniques based on vector integration enable monitoring of the fate of engineered cells in the blood of patients receiving GT and allow assessment of the safety and efficacy of these procedures. However, owing to the limited number of cells that can be tested and the impracticality of studying cells residing in peripheral organs without performing invasive biopsies, this approach provides only a partial snapshot of the clonal repertoire and dynamics of genetically modified cells and reduces the predictive power as a safety readout. In this study, we developed liquid biopsy integration site sequencing, or LiBIS-seq, a polymerase chain reaction technique optimized to quantitatively retrieve vector integration sites from cell-free DNA released into the bloodstream by dying cells residing in several tissues. This approach enabled longitudinal monitoring of in vivo liver-directed GT and clonal tracking in patients receiving hematopoietic stem cell GT, improving our understanding of the clonal composition and turnover of genetically modified cells in solid tissues and, in contrast to conventional analyses based only on circulating blood cells, enabling earlier detection of vector-marked clones that are aberrantly expanding in peripheral tissues.