Metabolic correction in oligodendrocytes derived from metachromatic leukodystrophy mouse model by using encapsulated recombinant myoblasts
Metabolic correction in oligodendrocytes derived from metachromatic leukodystrophy mouse model by using encapsulated recombinant myoblasts
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DOI：
10.1016/j.jns.2007.01.010
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发表时间：
2007-04-15
影响因子：
4.4
通讯作者：
Bordignon, Claudio
中科院分区：
文献类型：
--
作者：
Consiglio, Antonella;Martino, Sabata;Bordignon, Claudio
为了开发一种基于细胞的微囊化系统，将芳基硫酸酯酶A(ARSA)运送到异染性脑白质营养不良(MLD)患者的中枢神经系统，我们用含有全长人ARSA基因的逆转录病毒载体构建了C2C12小鼠成肌细胞，并评价了重组分泌酶逆转AS2(-/-)小鼠少突胶质细胞(OL)MLD表型的效果。转导后，C2C12细胞内ARSA活性增加15倍，ARSA分泌增加5倍。从聚醚-砜聚合物包裹的转导细胞中收集的hARSA被MLD小鼠来源的酶缺陷OL摄取，并正常分选到溶酶体室，在那里转移的酶达到生理水平的80%，恢复硫脂的代谢。为了评价分泌酶能否恢复脑内的代谢功能，体外实验证明囊化细胞和分泌型ARSA在脑脊液中是稳定的。此外，为了测试细胞活性和体内酶释放，将包裹的细胞移植到DBA/2J小鼠的背侧皮下。一个月后，所有回收的植入物释放hARSA的速度与未包裹的细胞相似，并含有保存完好的成肌细胞，表明包裹保持了C2C12细胞的分化、稳定的转基因表达和体内长期的细胞存活。因此，这些结果表明，基于聚合物包裹的转导异种细胞系用于MLD的基因治疗，开发ARSA向中枢神经系统的递送系统具有很好的潜力。(C)2007 Elsevier B.V.保留所有权利。
In an effort to develop an encapsulated cell-based system to deliver arylsulfatase A (ARSA) to the central nervous system of metachromatic leukodystrophy (MLD) patients, we engineered C2C12 mouse myoblasts with a retroviral vector containing a full-length human ARSA cDNA and evaluated the efficacy of the recombinant secreted enzyme to revert the MLD phenotype in oligodendrocytes (OL) of the As2(-/-) mouse model. After transduction, C2C12 cells showed a fifteen-fold increase in intracellular ARSA activity and five-fold increase in ARSA secretion. The secreted hARSA collected from transduced cells encapsulated in polyether-sulfone polymer, was taken up by enzyme-deficient OL derived from MLD mice and normally sorted to the lysosomal compartment, where transferred enzyme reached 80% of physiological levels, restoring the metabolism of sulfatide. To evaluate whether secreted enzyme could restore metabolic function in the brain, encapsulated cells and secreted ARSA were shown to be stable in CSF in vitro. Further, to test cell viability and enzyme release in vivo, encapsulated cells were implanted subcutaneously on the dorsal flank of DBA/2J mice. One month later, all retrieved implants released hARSA at rates similar to unencapsulated cells and contained well preserved myoblasts, demonstrating that encapsulation maintains differentiation Of C2C12 cells, stable transgene expression and long-term cell viability in vivo. Thus, these results show the promising potential of developing an ARSA delivery system to the CNS based on the use of a polymer-encapsulated transduced xenogenic cell line for gene therapy of MLD. (c) 2007 Elsevier B.V. All rights reserved.