Phenotypic correction of lipid storage and growth arrest in wolman disease fibroblasts by gene transfer of lysosomal acid lipase.
Phenotypic correction of lipid storage and growth arrest in wolman disease fibroblasts by gene transfer of lysosomal acid lipase.
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通过溶酶体酸性脂肪酶的基因转移对沃尔曼病成纤维细胞中脂质储存和生长停滞进行表型校正。
DOI：
10.1089/10430340150218413
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Rader,DJ
中科院分区：
文献类型：
--
作者：
Tietge,UJ;Sun,G;Czarnecki,S;Yu,Q;Lohse,P;Du,H;Grabowski,GA;Glick,JM;Rader,DJ
Wolman病是一种由于溶酶体酸性脂肪酶（LAL）缺乏而引起的致死性溶酶体贮积病。Wolman病的特征是明显的肝脏受累，而神经系统症状不常见， 这使得沃尔曼病成为肝脏定向基因治疗的一个有吸引力的候选者。本研究使用重组腺病毒在缺乏LAL的成纤维细胞中测试基因置换的效果。 编码人LAL cDNA（AdhLAL）。来自Wolman病患者的人成纤维细胞被AdhLAL感染，并且显示出LAL蛋白和活性的剂量依赖性增加，最高达5倍以上水平。 在对照成纤维细胞中。此外，72小时后感染AdhLAL有剂量依赖性校正严重的脂质储存表型的沃尔曼病成纤维细胞。电子显微镜证实 在超微结构水平上，AdhLAL感染的Wolman病成纤维细胞中溶酶体脂质储存的显著校正。将AdhLAL静脉注射到野生型小鼠中导致13.5倍的 肝LAL活性增加和LAL过表达与毒副作用无关。这些数据表明重组LALin vitroandin的高水平溶酶体表达 vivoand显示了基因治疗策略用于治疗Wolman病的可行性。
Wolman disease is a lethal lysosomal storage disease due to deficiency of lysosomal acid lipase (LAL). Wolman disease is characterized by pronounced hepatic involvement while neurological symptoms are uncommon, making Wolman disease an attractive candidate for liver-directed gene therapy. This study was performed to test the effects of gene replacement in fibroblasts lacking LAL, using a recombinant adenovirus encoding the human LAL cDNA (AdhLAL). Human fibroblasts from a Wolman disease patient were infected with AdhLAL and showed a dose-dependent increase in LAL protein and activity up to 5-fold above levels in control fibroblasts. Furthermore, 72 hr after infection with AdhLAL there was a dose-dependent correction of the severe lipid storage phenotype of Wolman disease fibroblasts. Electron microscopy confirmed significant correction of the lysosomal lipid storage in AdhLAL-infected Wolman disease fibroblasts at the ultrastructural level. Intravenous injection of AdhLAL into wild-type mice resulted in a 13.5-fold increase in hepatic LAL activity, and overexpression of LAL was not associated with toxic side effects. These data demonstrate high-level lysosomal expression of recombinant LALin vitroandin vivoand show the feasibility of gene therapeutic strategies for the treatment of Wolman disease.