Correction of a homoplasmic mitochondrial tRNA mutation in patient-derived iPSCs via a mitochondrial base editor

作者：Xiaoxu Chen, Mingyue Chen, Yuqing Zhu, Haifeng Sun, Yue Wang, Yuan Xie, Lianfu Ji, Cheng Wang, Zhibin Hu, Xuejiang Guo, Zhengfeng Xu, Jun Zhang, Shiwei Yang, Dong Liang, Bin Shen · 发表于：Communications Biology · 年份：2023 · DOI：10.1038/s42003-023-05500-y · 被引用次数：12 · 研究领域：Mitochondrial Function and Pathology、ATP Synthase and ATPases Research、CRISPR and Genetic Engineering

Pathogenic mutations in mitochondrial DNA cause severe and often lethal multi-system symptoms in primary mitochondrial defects. However, effective therapies for these defects are still lacking. Strategies such as employing mitochondrially targeted restriction enzymes or programmable nucleases to shift the ratio of heteroplasmic mutations and allotopic expression of mitochondrial protein-coding genes have limitations in treating mitochondrial homoplasmic mutations, especially in non-coding genes. Here, we conduct a proof of concept study applying a screened DdCBE pair to correct the homoplasmic m.A4300G mutation in induced pluripotent stem cells derived from a patient with hypertrophic cardiomyopathy. We achieve efficient G4300A correction with limited off-target editing, and successfully restore mitochondrial function in corrected induced pluripotent stem cell clones. Our study demonstrates the feasibility of using DdCBE to treat primary mitochondrial defects caused by homoplasmic pathogenic mitochondrial DNA mutations.