A Mutation-Agnostic Hematopoietic Stem Cell Gene Therapy for Metachromatic Leukodystrophy
A Mutation-Agnostic Hematopoietic Stem Cell Gene Therapy for Metachromatic Leukodystrophy
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DOI：
10.1089/crispr.2021.0075
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发表时间：
2021-12-08
期刊：
影响因子：
3.7
通讯作者：
Mezger, Markus
中科院分区：
文献类型：
--
作者：
Antony, Justin S.;Daniel-Moreno, Alberto;Mezger, Markus
异染性脑白质营养不良（MLD）是一种由芳基硫酸酯酶-A（ARSA）基因突变引起的罕见遗传性疾病。这种酶在脑细胞的硫苷脂代谢中起着关键作用，它的缺乏会导致神经退行性变。MLD的临床表现包括运动和认知功能的停滞和下降，导致标准治疗选择有限的过早死亡。在这里，我们描述了一种使用CRISPR-Cas9和AAV 6修复模板的突变不可知造血干细胞和祖细胞（HSPC）基因疗法，作为MLD的前瞻性治疗选择。我们的策略在人骨髓来源的HSPC中的ARSA位点实现了有效的插入和缺失（>87%）以及高水平的基因整合（>47%），没有可检测到的脱靶编辑。作为概念证明，我们在源自两名具有不同突变的MLD患者的HSPC中测试了我们的突变不可知疗法，并证明ARSA酶活性的恢复（>30倍改善）等同于健康成人。总之，我们的研究使MLD患者的突变不可知疗法具有经证实的疗效和强大的临床转化潜力。
Metachromatic leukodystrophy (MLD) is a rare genetic disorder caused by mutations in the Arylsulfatase-A (ARSA) gene. The enzyme plays a key role in sulfatide metabolism in brain cells, and its deficiency leads to neurodegeneration. The clinical manifestations of MLD include stagnation and decline of motor and cognitive function, leading to premature death with limited standard treatment options. Here, we describe a mutation-agnostic hematopoietic stem and progenitor cell (HSPC) gene therapy using CRISPR-Cas9 and AAV6 repair template as a prospective treatment option for MLD. Our strategy achieved efficient insertions and deletions (>87%) and a high level of gene integration (>47%) at the ARSA locus in human bone marrow-derived HSPCs, with no detectable off-target editing. As a proof of concept, we tested our mutation-agnostic therapy in HSPCs derived from two MLD patients with distinct mutations and demonstrated restoration of ARSA enzyme activity (>30-fold improvement) equivalent to healthy adults. In summary, our investigation enabled a mutation-agnostic therapy for MLD patients with proven efficacy and strong potential for clinical translation.