Netsawang Chaiwat, Tongbaen Malinee, Jearawiriyapaisarn Natee, Leecharoenkiat Kamonlak
Programme in Clinical Hematology Sciences, Department of Clinical Microscopy, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok, Thailand.
Thalassemia Research Center, Institute of Molecular Biosciences, Mahidol University, 25/25 Phuttamonthon 4 Road, Salaya, Nakhon Pathom, 73170, Thailand.
Sci Rep. 2025 Aug 18;15(1):30192. doi: 10.1038/s41598-025-15463-1.
Individuals with glucose-6-phosphate dehydrogenase (G6PD) deficiency hold a significant risk of severe hemolytic crises under oxidative stress. Currently, the definitive and curative treatment for the disorder has not been developed. Among over 200 G6PD variants, G6PD Viangchan (c.871 G > A) is the most prevalent and has been extensively studied in Southeast Asia. This study assessed the effectiveness of prime editing for correcting the G6PD Viangchan mutation in an established mutant HEK293T cell line and G6PD-deficient induced pluripotent stem cells (iPSCs). Using optimized modalities, prime editing achieved a high correction efficiency of over 25% in the HEK293T cells. In iPSCs, this gene editing tool yielded satisfactory correction outcomes, with approximately 5% corrected alleles. Our findings indicate that prime editing provides high precision, producing minimal by-products below baseline and showing undetectable off-target effects. Overall, prime editing has the potential to correct the G6PD Viangchan mutation, providing a valuable approach for future therapeutic strategies and the generation of isogenic cell lines to promote extensive studies in drug discovery and the pathogenesis of the G6PD variant.
葡萄糖-6-磷酸脱氢酶(G6PD)缺乏症患者在氧化应激下有发生严重溶血性危机的重大风险。目前,尚未开发出针对该疾病的确定性和治愈性治疗方法。在200多种G6PD变体中,G6PD万象(c.871 G > A)最为常见,并且在东南亚地区已得到广泛研究。本研究评估了碱基编辑在已建立的突变型HEK293T细胞系和G6PD缺陷诱导多能干细胞(iPSC)中校正G6PD万象突变的有效性。通过优化方法,碱基编辑在HEK293T细胞中实现了超过25%的高校正效率。在iPSC中,这种基因编辑工具产生了令人满意的校正结果,校正等位基因约为5%。我们的研究结果表明,碱基编辑具有高精度,产生的副产物低于基线水平且不可检测到脱靶效应。总体而言,碱基编辑有潜力校正G6PD万象突变,为未来的治疗策略以及生成同基因细胞系提供了一种有价值的方法,以促进在药物发现和G6PD变体发病机制方面的广泛研究。