Translational Immunology in Environmental Medicine, Klinikum Rechts der Isar, Technical University of Munich, Munich, Germany; Technical University of Munich, Munich, Germany.
Translational Immunology, Institute of Environmental Medicine, Helmholtz Zentrum Munich, Neuherberg, Germany; Technical University of Munich, Munich, Germany.
CRISPR J. 2021 Apr;4(2):178-190. doi: 10.1089/crispr.2020.0111.
STAT3-hyper IgE syndrome (STAT3-HIES) is a primary immunodeficiency presenting with destructive lung disease along with other symptoms. CRISPR-Cas9-mediated adenine base editors (ABEs) have the potential to correct one of the most common STAT3-HIES causing heterozygous mutations (c.1144C>T/p.R382W). As a proof-of-concept, we successfully applied ABEs to correct p.R382W in patient fibroblasts and induced pluripotent stem cells (iPSCs). Treated primary STAT3-HIES patient fibroblasts showed a correction efficiency of 29% ± 7% without detectable off-target effects evaluated through whole-genome and high-throughput sequencing. Compared with untreated patient fibroblasts, corrected single-cell clones showed functional rescue of STAT3 signaling with significantly increased STAT3 DNA-binding activity and target gene expression of and . Patient-derived iPSCs were corrected with an efficiency of 30% ± 6% and differentiated to alveolar organoids showing preserved plasticity in treated cells. In conclusion, our results are supportive for ABE-based gene correction as a potential causative treatment of STAT3-HIES.
STAT3 高免疫球蛋白 E 综合征（STAT3-HIES）是一种原发性免疫缺陷病，表现为破坏性肺部疾病以及其他症状。CRISPR-Cas9 介导的腺嘌呤碱基编辑器（ABEs）有可能纠正最常见的 STAT3-HIES 致病杂合突变之一（c.1144C>T/p.R382W）。作为概念验证，我们成功地将 ABEs 应用于纠正患者成纤维细胞和诱导多能干细胞（iPSCs）中的 p.R382W。经处理的原发性 STAT3-HIES 患者成纤维细胞的校正效率为 29%±7%，通过全基因组和高通量测序评估未发现可检测的脱靶效应。与未经处理的患者成纤维细胞相比，经校正的单细胞克隆显示 STAT3 信号的功能恢复，STAT3 DNA 结合活性和靶基因的表达显著增加。患者来源的 iPSCs 的校正效率为 30%±6%，并分化为肺泡类器官，显示出治疗细胞中保留的可塑性。总之，我们的结果支持基于 ABE 的基因校正作为 STAT3-HIES 的潜在因果治疗方法。
