作者
Wenyan Han,Hou-Yuan Qiu,S. F. Sun,Zhi-Can Fu,Guoquan Wang,Xiaowen Qian,Lijie Wang,Xiaowen Zhai,Jia Wang,Yichuan Wang,Yilin Guo,Guohua Cao,Rui-Jin Ji,Yizhou Zhang,Wei Wang,Hongsheng Wang,Mingli Zhao,Jing Wu,Lili Bi,Qiubing Chen,Zifeng Li,Ling Yu,Xiaozhou Mou,Hao Yin,Li Yang,Jia Chen,Bei Yang,Ying Zhang
摘要
Summary
Reactivating silenced γ-globin expression through the disruption of repressive regulatory domains offers a therapeutic strategy for treating β-hemoglobinopathies. Here, we used transformer base editor (tBE), a recently developed cytosine base editor with no detectable off-target mutations, to disrupt transcription-factor-binding motifs in hematopoietic stem cells. By performing functional screening of six motifs with tBE, we found that directly disrupting the BCL11A-binding motif in HBG1/2 promoters triggered the highest γ-globin expression. Via a side-by-side comparison with other clinical and preclinical strategies using Cas9 nuclease or conventional BEs (ABE8e and hA3A-BE3), we found that tBE-mediated disruption of the BCL11A-binding motif at the HBG1/2 promoters triggered the highest fetal hemoglobin in healthy and β-thalassemia patient hematopoietic stem/progenitor cells while exhibiting no detectable DNA or RNA off-target mutations. Durable therapeutic editing by tBE persisted in repopulating hematopoietic stem cells, demonstrating that tBE-mediated editing in HBG1/2 promoters is a safe and effective strategy for treating β-hemoglobinopathies.