Liver-directed gene therapy results in long-term correction of progressive familial intrahepatic cholestasis type 3 in mice
Liver-directed gene therapy results in long-term correction of progressive familial intrahepatic cholestasis type 3 in mice
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DOI：
10.1016/j.jhep.2019.03.021
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发表时间：
2019-07-01
影响因子：
25.7
通讯作者：
Bosma, Piter J.
中科院分区：
文献类型：
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作者：
Aronson, Sem J.;Bakker, Robert S.;Bosma, Piter J.
背景和目标：进行性家族性3型肝内胆汁淤积症（PFIC 3）的治疗选择有限，由于ABCB 4依赖性磷脂转运至胆汁受损，通常在成年前导致终末期肝病。使用腺相关病毒血清型8（AAV 8）介导的基因治疗，我们旨在将胆汁中的磷脂含量恢复到防止肝损伤的水平，从而使PFIC 3小鼠模型中的肝脏ABCB 4表达稳定并长期校正表型。10周龄的Abcb 4（-/-）小鼠在肝特异性启动子的控制下通过尾静脉注射接受单剂量的AAV 8-hABCB 4（n = 10）或AAV 8-GFP（n = 7）。在用0.1%胆酸盐饮食攻击2周后，在载体施用后10周或26周处死动物以评估转基因持久性。对血浆胆汁淤积标志物进行定期评价，胆管插管能够分析胆汁磷脂。免疫组化检测肝纤维化和Ki 67增殖指数。结果：给药后26周，所有接受AAV 8-hABCB 4的动物均获得稳定的转基因表达。AAV 8-hABCB 4表达恢复胆汁磷脂排泄，增加胆汁中的磷脂和胆固醇含量至改善肝损伤的水平。这导致血浆胆汁淤积标志物、碱性磷酸酶和胆红素正常化。此外，AAV 8-hABCB 4防止进行性肝纤维化和减少肝细胞增殖的持续时间的study.Conclusion：肝脏定向基因治疗提供了稳定的肝脏ABCB 4的表达和长期校正的表型在小鼠模型PFIC 3。进行性家族性肝内胆汁淤积症3型（PFIC 3）是一种严重的遗传性肝病，是由于脂质向胆汁的转运受损，使胆汁对肝细胞产生毒性而引起的。由于目前的治疗选择有限，本研究旨在评估基因治疗，以纠正这种疾病的小鼠模型中的潜在遗传缺陷。通过在小鼠肝细胞中引入缺失基因的功能性拷贝，我们能够恢复脂质向胆汁的转运，并大大减少对肝脏的损伤。肝细胞的增殖也减少，这有助于表型的长期校正。需要进一步的研究来评估这种方法是否可以应用于PFIC 3患者。(C)2019年欧洲肝脏研究协会。由爱思唯尔公司出版
Background & Aims: Progressive familial intrahepatic cholestasis type 3 (PFIC3), for which there are limited therapeutic options, often leads to end-stage liver disease before adulthood due to impaired ABCB4-dependent phospholipid transport to bile. Using adeno-associated virus serotype 8 (AAV8)-mediated gene therapy, we aimed to restore the phospholipid content in bile to levels that prevent liver damage, thereby enabling stable hepatic ABCB4 expression and long-term correction of the phenotype in a murine model of PFIC3.Methods: Ten-week-old Abcb4(-/-) mice received a single dose of AAV8-hABCB4 (n = 10) or AAV8-GFP (n = 7) under control of a liver specific promoter via tail vein injection. Animals were sacrificed either 10 or 26 weeks after vector administration to assess transgene persistence, after being challenged with a 0.1% cholate diet for 2 weeks. Periodic evaluation of plasma cholestatic markers was performed and bile duct cannulation enabled analysis of biliary phospholipids. Liver fibrosis and the Ki67 proliferation index were assessed by immunohistochemistry.Results: Stable transgene expression was achieved in all animals that received AAV8-hABCB4 up to 26 weeks after administration. AAV8-hABCB4 expression restored biliary phospholipid excretion, increasing the phospholipid and cholesterol content in bile to levels that ameliorate liver damage. This resulted in normalization of the plasma cholestatic markers, alkaline phosphatase and bilirubin. In addition, AAV8-hABCB4 prevented progressive liver fibrosis and reduced hepatocyte proliferation for the duration of the study.Conclusion: Liver-directed gene therapy provides stable hepatic ABCB4 expression and long-term correction of the phenotype in a murine model of PFIC3. Translational studies that verify the clinical feasibility of this approach are warranted.Lay summary: Progressive familial intrahepatic cholestasis type 3 (PFIC3) is a severe genetic liver disease that results from impaired transport of lipids to bile, which makes the bile toxic to liver cells. Because therapeutic options are currently limited, this study aims to evaluate gene therapy to correct the underlying genetic defect in a mouse model of this disease. By introducing a functional copy of the missing gene in liver cells of mice, we were able to restore lipid transport to bile and strongly reduce damage to the liver. The proliferation of liver cells was also reduced, which contributes to long-term correction of the phenotype. Further studies are required to evaluate whether this approach can be applied to patients with PFIC3. (C) 2019 European Association for the Study of the Liver. Published by Elsevier B.V.