Tao Rui, Xiao Lin, Zhou Lifang, Zheng Zhaoyue, Long Jie, Zhou Lixing, Tang Minghai, Dong Biao, Yao Shaohua
State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
National Clinical Research Center for Geriatrics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
Mol Ther Methods Clin Dev. 2020 Jan 13;19:507-517. doi: 10.1016/j.omtm.2019.12.014. eCollection 2020 Dec 11.
Phenylketonuria (PKU) is an inherited metabolic disorder caused by mutation within phenylalanine hydroxylase (PAH) gene. Loss-of-function of PAH leads to accumulation of phenylalanine in the blood/body of an untreated patient, which damages the developing brain, causing severe mental retardation. Current gene therapy strategies based on adeno-associated vector (AAV) delivery of PAH gene were effective in male animals but had little long-term effects on blood hyperphenylalaninemia in females. Here, we designed a gene therapy strategy using AAV to deliver a human codon-optimized phenylalanine amino lyase in a liver-specific manner. It was shown that PAL was active in lysing phenylalanine when it was expressed in mammalian cells. We produced a recombinant adeno-associated vector serotype 8 (AAV8) viral vector expressing the humanized PAL under the control of human antitrypsin (hAAT) promoter (AAV8-PAL). A single intravenous administration of AAV8-PAL caused long-term correction of hyperphenylalaninemia in both male and female PKU mice (strain Pah). Besides, no obvious liver injury was observed throughout the treatment process. Thus, our results established that AAV-mediated liver delivery of PAL gene is a promising strategy in the treatment of PKU.
苯丙酮尿症（PKU）是一种由苯丙氨酸羟化酶（PAH）基因突变引起的遗传性代谢紊乱疾病。PAH功能丧失会导致未经治疗的患者血液/体内苯丙氨酸积累，这会损害发育中的大脑，导致严重智力迟钝。目前基于腺相关病毒（AAV）递送PAH基因的基因治疗策略在雄性动物中有效，但对雌性动物的血液高苯丙氨酸血症几乎没有长期影响。在此，我们设计了一种基因治疗策略，使用AAV以肝脏特异性方式递送人密码子优化的苯丙氨酸氨基裂解酶。结果表明，当PAL在哺乳动物细胞中表达时，它在裂解苯丙氨酸方面具有活性。我们制备了一种重组腺相关病毒血清型8（AAV8）病毒载体，该载体在人抗胰蛋白酶（hAAT）启动子的控制下表达人源化PAL（AAV8-PAL）。单次静脉注射AAV8-PAL可使雄性和雌性PKU小鼠（Pah品系）的高苯丙氨酸血症得到长期纠正。此外，在整个治疗过程中未观察到明显的肝损伤。因此，我们的结果表明，AAV介导的肝脏递送PAL基因是治疗PKU的一种有前景的策略。
