In vivo genome editing of the albumin locus as a platform for protein replacement therapy
In vivo genome editing of the albumin locus as a platform for protein replacement therapy
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DOI：
10.1182/blood-2014-12-615492
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发表时间：
2015-10-08
期刊：
影响因子：
20.3
通讯作者：
High, Katherine A.
中科院分区：
文献类型：
--
作者：
Sharma, Rajiv;Anguela, Xavier M.;High, Katherine A.
定点基因组编辑为实现长期、稳定的治疗性基因表达提供了一种很有前途的方法。基因组编辑已经成功地应用于各种临床前模型，通常集中于靶向疾病基因座本身；然而，有限的靶向效率或来自内源启动子的表达不足可能阻碍这些方法的翻译，特别是如果所需的编辑事件不赋予选择性生长优势。在这里，我们报告了一个解决这些问题的肝脏导向蛋白替代疗法的一般策略：锌指核酸酶(ZFN)介导的白蛋白基因内治疗性转基因的定点整合。通过在体内使用腺相关病毒(AAV)载体，我们在血友病A和B小鼠模型中实现了人第VIII和第IX因子(hFVIII和hFIX)在治疗水平的长期表达。通过在野生型小鼠中使用相同的靶向试剂，表达了在Fabry和Gaucher病以及Hurler和Hunter综合征中缺乏的溶酶体酶。建立一个通用的基于核酸酶的分泌性蛋白生产平台将代表着在开发安全、永久和功能性治疗各种遗传性和非遗传性疾病方面的关键进展。
Site-specific genome editing provides a promising approach for achieving long-term, stable therapeutic gene expression. Genome editing has been successfully applied in a variety of preclinical models, generally focused on targeting the diseased locus itself; however, limited targeting efficiency or insufficient expression from the endogenous promoter may impede the translation of these approaches, particularly if the desired editing event does not confer a selective growth advantage. Here we report a general strategy for liver-directed protein replacement therapies that addresses these issues: zinc finger nuclease (ZFN) -mediated site-specific integration of therapeutic transgenes within the albumin gene. Byusing adeno-associated viral (AAV) vector delivery in vivo, we achieved long-term expression of human factors VIII and IX (hFVIII and hFIX) in mouse models of hemophilia A and B at therapeutic levels. By using the same targeting reagents in wild-type mice, lysosomal enzymes were expressed that are deficient in Fabry and Gaucher diseases and in Hurler and Hunter syndromes. The establishment of a universal nuclease-based platform for secreted protein production would represent a critical advance in the development of safe, permanent, and functional cures for diverse genetic and nongenetic diseases.