Department of Hematology and Oncology, University Children's Hospital, University of Tübingen, Germany.
Department of Pediatric Neurology, University Children's Hospital Tübingen, Germany.
CRISPR J. 2022 Feb;5(1):66-79. doi: 10.1089/crispr.2021.0075. Epub 2021 Dec 8.
Metachromatic leukodystrophy (MLD) is a rare genetic disorder caused by mutations in the () 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 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.
脑硫脂沉积病（MLD）是一种罕见的遗传疾病，由（）基因突变引起。该酶在脑细胞中硫酸脑苷脂代谢中发挥关键作用，其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，我们描述了一种基于 CRISPR-Cas9 和 AAV6 修复模板的无突变造血干细胞和祖细胞（HSPC）基因治疗，作为 MLD 的一种有前途的治疗选择。我们的策略在人骨髓来源的 HSPC 中实现了高效的插入和缺失（>87%）和高水平的基因整合（>47%）在 基因座上，没有检测到脱靶编辑。作为概念验证，我们在来自两名具有不同突变的 MLD 患者的 HSPC 中测试了我们的无突变治疗方法，并证明了 ARSA 酶活性的恢复（>30 倍的改善）与健康成年人相当。总之，我们的研究为 MLD 患者提供了一种经过验证的有效且具有很强临床转化潜力的无突变治疗方法。
