Phenylketonuria (PKU) is the most common inborn error of metabolism of the liver, and results from mutations of both alleles of the phenylalanine hydroxylase gene (PAH). As such, it is a suitable target for gene therapy via gene delivery with a recombinant adeno-associated virus (AAV) vector. Here we use the synthetic AAV vector Anc80 via systemic administration to deliver a functional copy of a codon-optimized human PAH gene, with or without an intron spacer, to the Pah^enu2 mouse model of PKU. Dose-dependent transduction of the liver and expression of PAH mRNA were present with both vectors, resulting in significant and durable reduction of circulating phenylalanine, reaching near control levels in males. Coat color of treated Pah^enu2 mice reflected an increase in pigmentation from brown to the black color of control animals, further indicating functional restoration of phenylalanine metabolism and its byproduct melanin. There were no adverse effects associated with administration of AAV up to 5 × 10¹² VG/kg, the highest dose tested. Only minor and/or transient variations in some liver enzymes were observed in some of the AAV-dosed animals which were not associated with pathology findings in the liver. Finally, there was no impact on cell turnover or apoptosis as evaluated by Ki-67 and TUNEL staining, further supporting the safety of this approach. This study demonstrates the therapeutic potential of AAV Anc80 to safely and durably cure PKU in a mouse model, supporting development for clinical consideration.

中文翻译：

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使用腺相关病毒血清型 Anc80 在小鼠模型中提供苯丙酮尿症的持久治愈

苯丙酮尿症 (PKU) 是最常见的先天性肝脏代谢错误，由苯丙氨酸羟化酶基因 ( PAH ) 的两个等位基因突变引起。因此，它是通过重组腺相关病毒 (AAV) 载体进行基因递送的基因治疗的合适靶标。在这里，我们通过全身给药使用合成的 AAV 载体 Anc80，将带有或不带有内含子间隔区的密码子优化的人类PAH基因的功能拷贝递送到 PKU 的Pah ^enu2小鼠模型。肝脏的剂量依赖性转导和PAH的表达两种载体都存在 mRNA，导致循环苯丙氨酸显着且持久地减少，在男性中达到接近控制水平。处理过的Pah ^enu2小鼠的毛色反映了对照动物从棕色到黑色的色素沉着增加，进一步表明苯丙氨酸代谢及其副产物黑色素的功能恢复。没有与施用 AAV 相关的不良反应高达 5 × 10 ¹²VG/kg，测试的最高剂量。在一些服用 AAV 的动物中仅观察到一些肝酶的轻微和/或短暂变化，这与肝脏中的病理学发现无关。最后，通过 Ki-67 和 TUNEL 染色评估，对细胞更新或细胞凋亡没有影响，进一步支持了这种方法的安全性。这项研究证明了 AAV Anc80 在小鼠模型中安全持久地治愈 PKU 的治疗潜力，支持临床考虑的开发。