Therapeutic potential of intracerebroventricular replacement of modified human β-hexosaminidase B for GM2 gangliosidosis.
Therapeutic potential of intracerebroventricular replacement of modified human β-hexosaminidase B for GM2 gangliosidosis.
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DOI：
10.1038/mt.2011.27
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发表时间：
2011-06
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--
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为了开发新的酶替代疗法治疗神经退行性Tay-Sachs病（TSD）和Sandhoff病（SD），我们设计了一个基因工程HEXB，编码嵌合的人β亚基，其中含有α亚基的部分氨基酸序列。我们成功地在中国仓鼠卵巢（CHO）细胞系稳定表达嵌合HEXB，它可以在体外降解人工阴离子底物和GM 2神经节苷脂，也保留了野生型（WT）HexB样热稳定性在血浆的存在下，产生修改的HexB。修饰的HexB通过阳离子非依赖性甘露糖6-磷酸受体有效地掺入来自Tay-Sachs患者的成纤维细胞中，并减少培养细胞中积累的GM 2神经节苷脂。此外，侧脑室给药的修改HexB山德霍夫模式小鼠恢复Hex的活动在大脑中，并减少了GM 2神经节苷脂储存在实质中。这些结果表明，侧脑室酶替代疗法涉及修改的HexB应该是更有效的Tay-Sachs和Sandhoff比利用HexA，特别是作为一个低抗原性的酶替代疗法的Tay-Sachs患者有内源性WT HexB。
To develop a novel enzyme replacement therapy for neurodegenerative Tay-Sachs disease (TSD) and Sandhoff disease (SD), which are caused by deficiency of β-hexosaminidase (Hex) A, we designed a genetically engineered HEXB encoding the chimeric human β-subunit containing partial amino acid sequence of the α-subunit by structure-based homology modeling. We succeeded in producing the modified HexB by a Chinese hamster ovary (CHO) cell line stably expressing the chimeric HEXB, which can degrade artificial anionic substrates and GM2 ganglioside in vitro, and also retain the wild-type (WT) HexB-like thermostability in the presence of plasma. The modified HexB was efficiently incorporated via cation-independent mannose 6-phosphate receptor into fibroblasts derived from Tay-Sachs patients, and reduced the GM2 ganglioside accumulated in the cultured cells. Furthermore, intracerebroventricular administration of the modified HexB to Sandhoff mode mice restored the Hex activity in the brains, and reduced the GM2 ganglioside storage in the parenchyma. These results suggest that the intracerebroventricular enzyme replacement therapy involving the modified HexB should be more effective for Tay-Sachs and Sandhoff than that utilizing the HexA, especially as a low-antigenic enzyme replacement therapy for Tay-Sachs patients who have endogenous WT HexB.
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