Preclinical modeling highlights the therapeutic potential of hematopoietic stem cell gene editing for correction of SCID-X1
Preclinical modeling highlights the therapeutic potential of hematopoietic stem cell gene editing for correction of SCID-X1
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DOI：
10.1126/scitranslmed.aan0820
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发表时间：
2017-10-11
影响因子：
17.1
通讯作者：
Naldini, Luigi
中科院分区：
文献类型：
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作者：
Schiroli, Giulia;Ferrari, Samuele;Naldini, Luigi
造血干细胞/祖细胞（HSPCs）靶向基因组编辑是治疗免疫血液病的一种有吸引力的策略。然而，在原始HSPCs中进行同源定向编辑的有限效率限制了校正细胞的产量，并可能影响临床翻译的可行性和安全性。这些问题需要在严格的临床前模型中得到解决，并通过开发更有效的编辑方法来克服。我们建立了一个人源化的x连锁严重联合免疫缺陷（SCID-X1）小鼠模型，并评估了从有限输入的功能性造血干细胞进行造血重建的有效性和安全性，建立了在不同类型条件作用下完全纠正的阈值。出乎意料的是，当移植少量祖细胞时，需要在HSPC输注前进行调节以保护小鼠免受淋巴瘤的发生。然后，我们设计了一种一刀切的il - 2rg（白细胞介素-2受体共同g链）基因校正策略，并使用适用于人类HSPC校正的相同试剂，在体内疾病模型中验证了编辑的人类基因，为小鼠HSPC靶向基因编辑提供了证据，并证明了il - 2rg编辑的淋巴细胞后代的功能。最后，我们优化了针对人类HSPC的编辑试剂和方案，并使用临床相关的HSPC来源和高度特异性的锌指核酸酶或CRISPR（聚集规律间隔的短回语重复序列）/Cas9 (CRISPR相关蛋白9)，达到了IL2RG在长期再生细胞中编辑的阈值，预计可以安全挽救疾病。总的来说，我们的工作为SCID-X1基因编辑的临床翻译奠定了理论基础和指导原则，并为开发其他疾病的基因校正提供了框架。
Targeted genome editing in hematopoietic stem/progenitor cells (HSPCs) is an attractive strategy for treating immunohematological diseases. However, the limited efficiency of homology-directed editing in primitive HSPCs constrains the yield of corrected cells and might affect the feasibility and safety of clinical translation. These concerns need to be addressed in stringent preclinical models and overcome by developing more efficient editing methods. We generated a humanized X-linked severe combined immunodeficiency (SCID-X1) mouse model and evaluated the efficacy and safety of hematopoietic reconstitution from limited input of functional HSPCs, establishing thresholds for full correction upon different types of conditioning. Unexpectedly, conditioning before HSPC infusion was required to protect the mice from lymphoma developing when transplanting small numbers of progenitors. We then designed a one-size-fits-all IL2RG (interleukin-2 receptor common g-chain) gene correction strategy and, using the same reagents suitable for correction of human HSPC, validated the edited human gene in the disease model in vivo, providing evidence of targeted gene editing in mouse HSPCs and demonstrating the functionality of the IL2RG-edited lymphoid progeny. Finally, we optimized editing reagents and protocol for human HSPCs and attained the threshold of IL2RG editing in long-term repopulating cells predicted to safely rescue the disease, using clinically relevant HSPC sources and highly specific zinc finger nucleases or CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9). Overall, our work establishes the rationale and guiding principles for clinical translation of SCID-X1 gene editing and provides a framework for developing gene correction for other diseases.