Mitigation of chromosome loss in clinical CRISPR-Cas9-engineered T cells.
Mitigation of chromosome loss in clinical CRISPR-Cas9-engineered T cells.
复制标题
DOI：
10.1016/j.cell.2023.08.041
复制
发表时间：
2023-10-12
期刊：
影响因子：
64.5
通讯作者：
Doudna, Jennifer A.
中科院分区：
文献类型：
--
作者：
Tsuchida, Connor A.;Brandes, Nadav;Bueno, Raymund;Trinidad, Marena;Mazumder, Thomas;Yu, Bingfei;Hwang, Byungjin;Chang, Christopher;Liu, Jamin;Sun, Yang;Hopkins, Caitlin R.;Parker, Kevin R.;Qi, Yanyan;Hofman, Laura;Satpathy, Ansuman T.;Stadtmauer, Edward A.;Cate, Jamie H. D.;Eyquem, Justin;Fraietta, Joseph A.;June, Carl H.;Chang, Howard Y.;Ye, Chun Jimmie;Doudna, Jennifer A.
CRISPR-Cas9基因组编辑已经使先进的T细胞疗法成为可能，但偶尔丢失靶向染色体仍然是一个安全问题。为了研究Cas9诱导的染色体丢失是否是一种普遍现象并评估其临床意义，我们对原代人类T细胞进行了系统的分析。排列和汇集的CRISPR筛查显示，染色体丢失在整个基因组中是普遍存在的，并导致目标染色体的部分和全部丢失，包括临床前嵌合抗原受体T细胞。染色体丢失的T细胞在培养中持续了数周，这意味着可能会干扰临床应用。在我们对Cas9工程T细胞(NCT03399448)进行的首个人类临床试验中，采用了一种改进的细胞制造工艺，在减少染色体丢失的同时，很大程度上保持了基因组编辑效率。P53的表达与该方案中观察到的防止染色体丢失相关，这表明T细胞工程在临床上减轻这种遗传毒性的机制和策略。CRISPR-Cas9基因组编辑会导致临床使用的人类原始T细胞和工程CAR T细胞持续的染色体丢失，这种现象可以在临床应用前通过修改T细胞制备方案来缓解。
CRISPR-Cas9 genome editing has enabled advanced T cell therapies, but occasional loss of the targeted chromosome remains a safety concern. To investigate whether Cas9-induced chromosome loss is a universal phenomenon and evaluate its clinical significance, we conducted a systematic analysis in primary human T cells. Arrayed and pooled CRISPR screens revealed that chromosome loss was generalizable across the genome and resulted in partial and entire loss of the targeted chromosome, including in preclinical chimeric antigen receptor T cells. T cells with chromosome loss persisted for weeks in culture, implying the potential to interfere with clinical use. A modified cell manufacturing process, employed in our first-in-human clinical trial of Cas9-engineered T cells (NCT03399448), reduced chromosome loss while largely preserving genome editing efficacy. Expression of p53 correlated with protection from chromosome loss observed in this protocol, suggesting both a mechanism and strategy for T cell engineering that mitigates this genotoxicity in the clinic. CRISPR-Cas9 genome editing induces persistent chromosome loss in human primary T cells and engineered CAR T cells for clinical use, a phenomenon that can be mitigated by modifying T cell preparation protocols prior to clinical application.
登录
查看更多内容
影响因子：
120.1
作者：
Foy, Susan P. P.;Jacoby, Kyle;Bota, Daniela A. A.;Hunter, Theresa;Pan, Zheng;Stawiski, Eric;Ma, Yan;Lu, William;Peng, Songming;Wang, Clifford L. L.;Yuen, Benjamin;Dalmas, Olivier;Heeringa, Katharine;Sennino, Barbara;Conroy, Andy;Bethune, Michael T. T.;Mende, Ines;White, William;Kukreja, Monica;Gunturu, Swetha;Humphrey, Emily;Hussaini, Adeel;An, Duo;Litterman, Adam J. J.;Quach, Boi Bryant;Ng, Alphonsus H. C.;Lu, Yue;Smith, Chad;Campbell, Katie M. M.;Anaya, Daniel;Skrdlant, Lindsey;Huang, Eva Yi-Hsuan;Mendoza, Ventura;Mathur, Jyoti;Dengler, Luke;Purandare, Bhamini;Moot, Robert;Yi, Michael C. C.;Funke, Roel;Sibley, Alison;Stallings-Schmitt, Todd;Oh, David Y. Y.;Chmielowski, Bartosz;Abedi, Mehrdad;Yuan, Yuan;Sosman, Jeffrey A. A.;Lee, Sylvia M. M.;Schoenfeld, Adam J. J.;Baltimore, David;Heath, James R. R.;Franzusoff, Alex;Ribas, Antoni;Rao, Arati V. V.;Mandl, Stefanie J. J.
通讯作者：
Mandl, Stefanie J. J.
影响因子：
64.8
作者：
Eyquem J;Mansilla-Soto J;Giavridis T;van der Stegen SJ;Hamieh M;Cunanan KM;Odak A;Gönen M;Sadelain M
通讯作者：
Sadelain M
影响因子：
17.1
作者：
Brentjens RJ;Davila ML;Riviere I;Park J;Wang X;Cowell LG;Bartido S;Stefanski J;Taylor C;Olszewska M;Borquez-Ojeda O;Qu J;Wasielewska T;He Q;Bernal Y;Rijo IV;Hedvat C;Kobos R;Curran K;Steinherz P;Jurcic J;Rosenblat T;Maslak P;Frattini M;Sadelain M
通讯作者：
Sadelain M
影响因子：
7
作者：
Kim S;Kim D;Cho SW;Kim J;Kim JS
通讯作者：
Kim JS
影响因子：
64.8
作者：
Dever DP;Bak RO;Reinisch A;Camarena J;Washington G;Nicolas CE;Pavel-Dinu M;Saxena N;Wilkens AB;Mantri S;Uchida N;Hendel A;Narla A;Majeti R;Weinberg KI;Porteus MH
通讯作者：
Porteus MH