Yin Shuming, Gao Liangcai, Sun Xiaoyue, Zhang Mei, Gao Hongyi, Chen Xiaoqing, Zhang Dan, Ming Xinyu, Yang Lei, Hu Yaqiang, Chen Xi, Liu Meizhen, Zhan Xia, Guan Yuting, Wang Liren, Han Lianshu, Zhu Ping, Li Dali
Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Southern Medical University, Guangzhou, Guangdong 510100, China; Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai 200241, China; School of Life Sciences, East China Normal University, Shanghai 200241, China.
School of Life Sciences, East China Normal University, Shanghai 200241, China.
Mol Ther. 2025 Jan 8;33(1):119-132. doi: 10.1016/j.ymthe.2024.11.032. Epub 2024 Nov 26.
Phenylketonuria (PKU) is a liver metabolic disorder mainly caused by a deficiency of the hepatic phenylalanine hydroxylase (PAH) enzyme activity, often leading to severe brain function impairment in patients if untreated or if treatment is delayed. In this study, we utilized dual-AAV8 vectors to deliver a near PAM-less adenine base editor variant, known as ABE8e-SpRY, to treat the Pah PKU mouse model carrying a frequent R408W mutation in the Pah gene. Our findings revealed that a single intravenous injection in adult mice and a single intraperitoneal injection in neonatal mice resulted in 19.1%-34.6% A-to-G editing efficiency at the pathogenic mutation site with minimal bystander edits. Furthermore, the dual-AAV8-treated mice exhibited reduced blood Phe levels to below the therapeutic threshold of 360 μmol L and restored weight and fur color to normal levels. Importantly, the brain function of the mice was restored after the treatment, particularly when administered during the neonatal stage, as levels of monoamine neurotransmitters and metabolites in the brain returned to normal and near-normal levels. Our study demonstrated that ABE8e-SpRY-based base editing could effectively correct the point mutation in the Pah PKU mouse model, indicating potential clinical applications for PKU and other genetic diseases.
苯丙酮尿症（PKU）是一种肝脏代谢紊乱疾病，主要由肝脏苯丙氨酸羟化酶（PAH）酶活性缺乏引起，如果不治疗或治疗延迟，通常会导致患者严重的脑功能损害。在本研究中，我们利用双AAV8载体递送一种近乎无PAM的腺嘌呤碱基编辑器变体，即ABE8e-SpRY，来治疗携带Pah基因常见R408W突变的Pah PKU小鼠模型。我们的研究结果表明，在成年小鼠中进行单次静脉注射以及在新生小鼠中进行单次腹腔注射，在致病突变位点产生了19.1%-34.6%的A到G编辑效率，且旁观者编辑极少。此外，经双AAV8处理的小鼠血液苯丙氨酸水平降至360 μmol/L的治疗阈值以下，体重和毛色恢复到正常水平。重要的是，治疗后小鼠的脑功能得到恢复，尤其是在新生阶段给药时，因为大脑中的单胺神经递质和代谢物水平恢复到正常和接近正常水平。我们的研究表明，基于ABE8e-SpRY的碱基编辑可以有效纠正Pah PKU小鼠模型中的点突变，这表明PKU和其他遗传疾病具有潜在的临床应用价值。
