Life-Long Correction of Hyperbilirubinemia with a Neonatal Liver-Specific AAV-Mediated Gene Transfer in a Lethal Mouse Model of Crigler-Najjar Syndrome
Life-Long Correction of Hyperbilirubinemia with a Neonatal Liver-Specific AAV-Mediated Gene Transfer in a Lethal Mouse Model of Crigler-Najjar Syndrome
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DOI：
10.1089/hum.2013.233
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发表时间：
2014-09-01
影响因子：
4.2
通讯作者：
Muro, Andres F.
中科院分区：
文献类型：
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作者：
Bortolussi, Giulia;Zentillin, Lorena;Muro, Andres F.
UGT 1A 1基因的突变导致Crigler-Najjar综合征I型（CNSI），其特征是严重的高胆红素血症和发生神经损伤的持续风险。光疗治疗降低血浆胆红素水平，但其疗效有限，需要进行肝移植。为了找到替代疗法，我们将AAV肝脏特异性基因疗法应用于CNSI的致死小鼠模型。我们证明，一个单一的新生儿hUGT 1A 1基因转移是成功的，治疗效果持续到17个月后注射。治疗效果是由转录活性双链附加体介导的。我们还比较了两种不同的基因治疗方法的疗效：肝脏与骨骼肌转基因表达。我们观察到，正常肝脏表达和活性水平的5-8%足以显著降低胆红素水平并维持终身低血浆胆红素浓度（3.1 +/- 1.5 mg/dl）。相比之下，骨骼肌不能有效地降低胆红素（6.4 +/- 2.0 mg/dl），尽管与正常肝脏相比，hUgt 1a 1表达水平为20-30%。我们认为，这种基因治疗效果的显着差异可能与肌肉中结合胆红素的Mrp 2和Mrp 3转运蛋白的缺乏有关。综上所述，我们的数据支持肝脏是有效和长期CNSI基因治疗的最佳器官的概念，并建议肝外组织的使用应与胆红素转运蛋白的存在相结合。
Null mutations in the UGT1A1 gene result in Crigler-Najjar syndrome type I (CNSI), characterized by severe hyperbilirubinemia and constant risk of developing neurological damage. Phototherapy treatment lowers plasma bilirubin levels, but its efficacy is limited and liver transplantation is required. To find alternative therapies, we applied AAV liver-specific gene therapy to a lethal mouse model of CNSI. We demonstrated that a single neonatal hUGT1A1 gene transfer was successful and the therapeutic effect lasted up to 17 months postinjection. The therapeutic effect was mediated by the presence of transcriptionally active double-stranded episomes. We also compared the efficacy of two different gene therapy approaches: liver versus skeletal muscle transgene expression. We observed that 5-8% of normal liver expression and activity levels were sufficient to significantly reduce bilirubin levels and maintain lifelong low plasma bilirubin concentration (3.1 +/- 1.5 mg/dl). In contrast, skeletal muscle was not able to efficiently lower bilirubin (6.4 +/- 2.0 mg/dl), despite 20-30% of hUgt1a1 expression levels, compared with normal liver. We propose that this remarkable difference in gene therapy efficacy could be related to the absence of the Mrp2 and Mrp3 transporters of conjugated bilirubin in muscle. Taken together, our data support the concept that liver is the best organ for efficient and long-term CNSI gene therapy, and suggest that the use of extra-hepatic tissues should be coupled to the presence of bilirubin transporters.