Therapeutic gene editing in CD34(+) hematopoietic progenitors from Fanconi anemia patients.
Therapeutic gene editing in CD34(+) hematopoietic progenitors from Fanconi anemia patients.
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DOI：
10.15252/emmm.201707540
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发表时间：
2017-11
影响因子：
11.1
通讯作者：
Rio P
中科院分区：
文献类型：
--
作者：
Diez B;Genovese P;Roman-Rodriguez FJ;Alvarez L;Schiroli G;Ugalde L;Rodriguez-Perales S;Sevilla J;Diaz de Heredia C;Holmes MC;Lombardo A;Naldini L;Bueren JA;Rio P
基因打靶是遗传性疾病基因治疗发展的新阶段。尽管先前的研究已经显示了编辑范可尼贫血（FA）患者的成纤维细胞的可行性，但在这里，我们的目标是在临床相关细胞（如造血干细胞（HSC））中进行治疗性基因编辑。在我们的第一个实验中，我们表明锌指核酸酶（ZFN）介导的非治疗性EGFP报告供体插入FA-A淋巴母细胞系（LCL）的AAVS 1“安全港”基因座，表明FANCA对于编辑人类细胞不是必需的。当用治疗性FANCA供体进行相同的方法时，获得了FA-A LCL的有效表型校正。使用来自健康供体的原代脐带血CD 34+细胞，不仅在体外培养的细胞中，而且在负责原代和继发性免疫缺陷小鼠的再增殖的造血前体中证实了基因靶向。此外，当使用来自FA‐A患者的动员外周血CD 34+细胞进行类似实验时，我们可以首次证明来自FA患者的初级造血前体中的基因靶向是可行的，并且与这些临床相关细胞的表型校正相容。
Gene targeting constitutes a new step in the development of gene therapy for inherited diseases. Although previous studies have shown the feasibility of editing fibroblasts from Fanconi anemia (FA) patients, here we aimed at conducting therapeutic gene editing in clinically relevant cells, such as hematopoietic stem cells (HSCs). In our first experiments, we showed that zinc finger nuclease (ZFN)‐mediated insertion of a non‐therapeutic EGFP‐reporter donor in the AAVS1 “safe harbor” locus of FA‐A lymphoblastic cell lines (LCLs), indicating that FANCA is not essential for the editing of human cells. When the same approach was conducted with therapeutic FANCA donors, an efficient phenotypic correction of FA‐A LCLs was obtained. Using primary cord blood CD34+ cells from healthy donors, gene targeting was confirmed not only in in vitro cultured cells, but also in hematopoietic precursors responsible for the repopulation of primary and secondary immunodeficient mice. Moreover, when similar experiments were conducted with mobilized peripheral blood CD34+ cells from FA‐A patients, we could demonstrate for the first time that gene targeting in primary hematopoietic precursors from FA patients is feasible and compatible with the phenotypic correction of these clinically relevant cells.
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影响因子：
64.8
作者：
通讯作者：
--
影响因子：
64.8
作者：
Genovese, Pietro;Schiroli, Giulia;Escobar, Giulia;Di Tomaso, Tiziano;Firrito, Claudia;Calabria, Andrea;Moi, Davide;Mazzieri, Roberta;Bonini, Chiara;Holmes, Michael C.;Gregory, Philip D.;van der Burg, Mirjam;Gentner, Bernhard;Montini, Eugenio;Lombardo, Angelo;Naldini, Luigi
通讯作者：
Naldini, Luigi
影响因子：
20.3
作者：
BUTTURINI, A;GALE, RP;AUERBACH, AD
通讯作者：
AUERBACH, AD
影响因子：
23.9
作者：
Ceccaldi, Raphael;Parmar, Kalindi;Mouly, Enguerran;Delord, Marc;Kim, Jung Min;Regairaz, Marie;Pla, Marika;Vasquez, Nadia;Zhang, Qing-Shuo;Pondarre, Corinne;de Latour, Regis Peffault;Gluckman, Eliane;Cavazzana-Calvo, Marina;Leblanc, Thierry;Larghero, Jerome;Grompe, Markus;Socie, Gerard;D'Andrea, Alan D.;Soulier, Jean
通讯作者：
Soulier, Jean
影响因子：
5.1
作者：
Charrier, S.;Ferrand, M.;Galy, A.
通讯作者：
Galy, A.