In vivo HSC prime editing rescues sickle cell disease in a mouse model.
In vivo HSC prime editing rescues sickle cell disease in a mouse model.
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体内 HSC Prime 编辑可挽救小鼠模型中的镰状细胞病。
DOI：
10.1182/blood.2022018252
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发表时间：
2023-04-27
期刊：
影响因子：
20.3
通讯作者：
Lieber, Andre
中科院分区：
文献类型：
--
作者：
Li, Chang;Georgakopoulou, Aphrodite;Newby, Gregory A.;Chen, Peter J.;Everette, Kelcee A.;Paschoudi, Kiriaki;Vlachaki, Efthymia;Gil, Sucheol;Anderson, Anna K.;Koob, Theodore;Huang, Lishan;Wang, Hongjie;Kiem, Hans-Peter;Liu, David R.;Yannaki, Evangelia;Lieber, Andre
镰状细胞突变和疾病表型的纠正是通过体内HSC转导与矢量化的初始编辑器实现的。我们的体内HSC引物编辑方法不需要HSC移植和骨髓消融，应该简化HSC基因治疗。镰状细胞病（SCD）是一种由β-珠蛋白基因核苷酸突变引起的单基因疾病。目前的基因治疗研究主要集中在慢病毒载体介导的基因添加或CRISPR/ cas9介导的胎儿珠蛋白再激活，根本原因没有得到解决。我们在SCD小鼠模型（CD46/Townes小鼠）中开发了一种矢量化的先导编辑系统，可以直接修复造血干细胞（hsc）中的SCD突变。我们的方法包括单次静脉注射一种非整合的、主要编辑器表达的病毒载体到动员的CD46/Townes小鼠中，并在体内进行低剂量药物选择。该方法校正了造血干细胞中约40%的βS等位基因。平均43%的镰状血红蛋白被成人血红蛋白取代，从而大大减轻了SCD表型。继发受体的移植表明，长期再生的造血干细胞是经过编辑的。以最小的插入和删除生成实现了高效的目标位点编辑，并且没有检测到脱靶编辑。由于其简单性和可移植性，我们的体内初始编辑方法具有在SCD普遍存在的资源贫乏国家应用的潜力。李和他的同事报道了一种治疗镰状细胞病的新基因疗法。与慢病毒基因添加或CRISPR/ cas9介导的胎儿血红蛋白再激活的体外操作不同，作者描述了将非整合的主要编辑表达载体注射到镰状小鼠模型中，在体内校正了40%以上的血红蛋白S等位基因。虽然有几个特点需要优化，但这项技术为在资源贫乏的环境中进行基因治疗提供了潜力。
Correction of the sickle-cell mutation and disease phenotypes is achieved by in vivo HSC transduction with vectorized prime editors. Our approach for in vivo HSC prime editing that does not require HSC transplantation and myeloablation should simplify HSC gene therapy. Sickle cell disease (SCD) is a monogenic disease caused by a nucleotide mutation in the β-globin gene. Current gene therapy studies are mainly focused on lentiviral vector–mediated gene addition or CRISPR/Cas9–mediated fetal globin reactivation, leaving the root cause unfixed. We developed a vectorized prime editing system that can directly repair the SCD mutation in hematopoietic stem cells (HSCs) in vivo in a SCD mouse model (CD46/Townes mice). Our approach involved a single intravenous injection of a nonintegrating, prime editor–expressing viral vector into mobilized CD46/Townes mice and low-dose drug selection in vivo. This procedure resulted in the correction of ∼40% of βS alleles in HSCs. On average, 43% of sickle hemoglobin was replaced by adult hemoglobin, thereby greatly mitigating the SCD phenotypes. Transplantation in secondary recipients demonstrated that long-term repopulating HSCs were edited. Highly efficient target site editing was achieved with minimal generation of insertions and deletions and no detectable off-target editing. Because of its simplicity and portability, our in vivo prime editing approach has the potential for application in resource-poor countries where SCD is prevalent. Li and colleagues report on a novel gene-therapy approach to sickle cell disease. Rather than ex vivo manipulation with lentiviral gene addition or CRISPR/Cas9-mediated fetal hemoglobin reactivation, the authors describe injection of a nonintegrating prime editor–expressing vector into a sickle mouse model with correction of over 40% of hemoglobin S alleles in vivo. Though several features need to be optimized, this technique offers a potential for gene-therapy delivery in resource-poor settings.
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影响因子：
64.8
作者：
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Naldini, Luigi
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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
影响因子：
64.8
作者：
Anzalone, Andrew V.;Randolph, Peyton B.;Liu, David R.
通讯作者：
Liu, David R.