Reprogramming human T cell function and specificity with non-viral genome targeting.
Reprogramming human T cell function and specificity with non-viral genome targeting.
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DOI：
10.1038/s41586-018-0326-5
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发表时间：
2018-07
期刊：
影响因子：
64.8
通讯作者：
Marson A
中科院分区：
文献类型：
--
作者：
Roth TL;Puig-Saus C;Yu R;Shifrut E;Carnevale J;Li PJ;Hiatt J;Saco J;Krystofinski P;Li H;Tobin V;Nguyen DN;Lee MR;Putnam AL;Ferris AL;Chen JW;Schickel JN;Pellerin L;Carmody D;Alkorta-Aranburu G;Del Gaudio D;Matsumoto H;Morell M;Mao Y;Cho M;Quadros RM;Gurumurthy CB;Smith B;Haugwitz M;Hughes SH;Weissman JS;Schumann K;Esensten JH;May AP;Ashworth A;Kupfer GM;Greeley SAW;Bacchetta R;Meffre E;Roncarolo MG;Romberg N;Herold KC;Ribas A;Leonetti MD;Marson A
几十年来，人们一直致力于利用重组病毒载体对T细胞进行基因重编程以用于治疗目的，这些病毒载体不会将转基因靶向特定的基因组位点。此外，对病毒载体的需求减缓了研究和临床应用，因为它们的生产和检测过程漫长且昂贵。基因组编辑带来了通过同源定向修复（HDR）将大型转基因特异性且高效地插入靶细胞的希望。在此，我们开发了一种不需要病毒载体的CRISPR - Cas9基因组靶向系统，该系统能够在原代人T细胞基因组的特定位点快速且高效地插入大的DNA序列（>1kb），同时保持细胞的活力和功能。这允许对内源基因进行单个或多重修饰。首先，我们应用这一策略纠正单基因自身免疫疾病患者细胞中的致病性IL2RA突变，证明其信号功能得到改善。其次，我们用一种新的T细胞受体（TCR）替换内源性T细胞受体位点，使T细胞重新定向到一种癌症抗原。由此产生的经TCR工程改造的T细胞能够特异性识别肿瘤抗原，并在体外和体内产生有效的抗肿瘤细胞反应。综上所述，这些研究提供了临床前证据，表明非病毒基因组靶向能够对原代人免疫细胞进行快速且灵活的实验操作和治疗性工程改造。
Decades of work have aimed to genetically reprogram T cells for therapeutic purposes using recombinant viral vectors, which do not target transgenes to specific genomic sites. In addition, the need for viral vectors has slowed down research and clinical use as their manufacturing and testing is lengthy and expensive. Genome editing brought the promise of specific and efficient insertion of large transgenes into target cells through homology-directed repair (HDR). Here, we developed a CRISPR-Cas9 genome targeting system that does not require viral vectors, allowing rapid and efficient insertion of large DNA sequences (> 1kb) at specific sites in the genomes of primary human T cells, while preserving cell viability and function. This permits individual or multiplexed modification of endogenous genes. First, we apply this strategy to correct a pathogenic IL2RA mutation in cells from patients with monogenic autoimmune disease, demonstrating improved signaling function. Second, we replace the endogenous T cell receptor (TCR) locus with a new TCR redirecting T cells to a cancer antigen. The resulting TCR-engineered T cells specifically recognize tumour antigen and mount productive anti-tumour cell responses in vitro and in vivo. Taken together, these studies provide preclinical evidence that non-viral genome targeting can enable rapid and flexible experimental manipulation and therapeutic engineering of primary human immune cells.
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影响因子：
12.3
作者：
Quadros RM;Miura H;Harms DW;Akatsuka H;Sato T;Aida T;Redder R;Richardson GP;Inagaki Y;Sakai D;Buckley SM;Seshacharyulu P;Batra SK;Behlke MA;Zeiner SA;Jacobi AM;Izu Y;Thoreson WB;Urness LD;Mansour SL;Ohtsuka M;Gurumurthy CB
通讯作者：
Gurumurthy CB
影响因子：
48
作者：
Mali, Prashant;Esvelt, Kevin M.;Church, George M.
通讯作者：
Church, George M.
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：
64.8
作者：
Simeonov DR;Gowen BG;Boontanrart M;Roth TL;Gagnon JD;Mumbach MR;Satpathy AT;Lee Y;Bray NL;Chan AY;Lituiev DS;Nguyen ML;Gate RE;Subramaniam M;Li Z;Woo JM;Mitros T;Ray GJ;Curie GL;Naddaf N;Chu JS;Ma H;Boyer E;Van Gool F;Huang H;Liu R;Tobin VR;Schumann K;Daly MJ;Farh KK;Ansel KM;Ye CJ;Greenleaf WJ;Anderson MS;Bluestone JA;Chang HY;Corn JE;Marson A
通讯作者：
Marson A
影响因子：
7
作者：
Kim S;Kim D;Cho SW;Kim J;Kim JS
通讯作者：
Kim JS