Niemann–Pick disease type C1 (NPC1) is a fatal congenital neurodegenerative disorder caused by mutations in the NPC1 gene, which is involved in cholesterol transport in lysosomes. Broad clinical manifestations of NPC1 include liver failure, pulmonary disorder, neurological deficits, and psychiatric symptoms. The main cause of death in NPC1 patients involves central nervous system (CNS) dysfunction; there is no essential treatment. We generated a tyrosine-mutant adeno-associated virus (AAV) 9/3 vector that expresses human NPC1 under a cytomegalovirus (CMV) promoter (AAV-CMV-hNPC1) and injected it into the left lateral ventricle (5 μL) and cisterna magna (10 μL) of Npc1 homo-knockout (Npc1^−/−) mice. Each mouse received total 1.35 × 10¹¹ vector genome on days 4 or 5 of life. AAV-treated Npc1^−/− mice (n = 11) had an average survival of >28 weeks, while all saline-treated Npc1^−/− mice (n = 11) and untreated Npc1^−/− mice (n = 6) died within 16 weeks. Saline-treated and untreated Npc1^−/− mice lost body weight from 7 weeks until death. However, the average body weight of AAV-treated Npc1^−/− mice increased until 15 weeks. AAV-treated Npc1^−/− mice also showed a significant improvement in the rotarod test performance. A pathological analysis at 11 weeks showed that cerebellar Purkinje cells were preserved in AAV-treated Npc1^−/− mice. In contrast, untreated Npc1^−/− mice showed an almost total loss of cerebellar Purkinje cells. Combined injection into both the lateral ventricle and cisterna magna achieved broader delivery of the vector to the CNS, leading to better outcomes than noted in previous reports, with injection into the lateral ventricles or veins alone. In AAV-treated Npc1^−/− mice, vector genome DNA was detected widely in the CNS and liver. Human NPC1 RNA was detected in the brain, liver, lung, and heart. Accumulated unesterified cholesterol in the liver was reduced in the AAV-treated Npc1^−/− mice. Our results suggest the feasibility of gene therapy for patients with NPC1.

中文翻译：

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尼曼-匹克病 C1 型小鼠模型中的基因治疗

Niemann-Pick C1 型 (NPC1) 是一种致命的先天性神经退行性疾病，由NPC1基因突变引起，该基因与溶酶体中的胆固醇转运有关。NPC1 的广泛临床表现包括肝功能衰竭、肺部疾病、神经功能缺损和精神症状。NPC1 患者的主要死亡原因是中枢神经系统（CNS）功能障碍；没有必要的治疗。我们生成了在巨细胞病毒 (CMV) 启动子 (AAV-CMV- hNPC1 ) 下表达人NPC1的酪氨酸突变腺相关病毒 (AAV) 9/3 载体，并将其注射到左侧脑室 (5 μL) 和小脑池(10 μL) Npc1同源基因敲除 ( Npc1 ^{- /-} ) 老鼠。每只小鼠在生命的第 4 天或第 5 天收到总共 1.35 × 10 ^{11 个}载体基因组。AAV 处理的Npc1 ^{- / -}小鼠 ( n  = 11) 的平均存活时间 > 28 周，而所有盐水处理的Npc1 ^{- / -}小鼠 ( n  = 11) 和未处理的 Npc1 ^{- / -}小鼠 ( n  = 6) 均死亡16 周内。盐水处理和未处理的Npc1 ^{- / -}小鼠从 7 周开始体重下降直至死亡。然而，AAV 处理的Npc1的平均体重^{- / -}小鼠增加至 15 周。AAV 处理的Npc1 ^{- / -}小鼠也显示出旋转棒测试性能的显着改善。11 周时的病理学分析表明，小脑浦肯野细胞在 AAV 处理的Npc1 ^{- / -}小鼠中得以保存。相反，未经处理的 Npc1 ^{- / -}小鼠显示小脑浦肯野细胞几乎完全丧失。侧脑室和小脑池的联合注射实现了将载体更广泛地传递到中枢神经系统，导致比以前报告中提到的更好的结果，单独注射到侧脑室或静脉中。在 AAV 处理的Npc1 ^{- /−}小鼠，在 CNS 和肝脏中广泛检测到载体基因组 DNA。在大脑、肝脏、肺和心脏中检测到人NPC1 RNA。在 AAV 处理的Npc1 ^{- / -}小鼠中，肝脏中累积的未酯化胆固醇减少。我们的结果表明基因治疗对 NPC1 患者的可行性。