A human neuronal tissue culture model for Lesch-Nyhan disease
A human neuronal tissue culture model for Lesch-Nyhan disease
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DOI：
10.1111/j.1471-4159.2007.04472.x
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发表时间：
2007-05-01
影响因子：
4.7
通讯作者：
Jinnah, H. A.
中科院分区：
文献类型：
--
作者：
Shirley, Thomas L.;Lewers, J. Chris;Jinnah, H. A.
编码嘌呤补救酶（次黄嘌呤-鸟嘌呤磷酸核糖转移酶（HPRT））的基因突变导致莱-尼二氏病，这是一种以认知、神经和行为异常为特征的神经发育障碍。尽管对酶的功能有详细的了解，但伴随嘌呤再循环丧失的关键病理生理变化尚不清楚。为了便于描述HPRT缺陷的后果，通过三螺旋形成寡核苷酸的靶向诱变分离了四个独立的人多巴胺能神经母细胞瘤的HPRT缺陷亚系SK-N-BE（2）M17。作为一个组，这些HPRT缺陷细胞显示出几种显著的异常：（i）嘌呤再循环受损，伴随次黄嘌呤、鸟嘌呤和黄嘌呤的积累，（ii）鸟苷酸能荷和GTP减少：GDP比率，但腺苷酸能荷正常，任何腺嘌呤核苷酸比率无变化，（iii）UTP和NADP（+）水平升高，（iv）多巴脱羧酶降低，但单胺正常，和（v）细胞索马大小的减小。这些细胞联合收割机结合了多个细胞系的分析能力和人类神经元来源，为研究HPRT缺陷的病理生理学提供了重要工具。
Mutations in the gene encoding the purine salvage enzyme, hypoxanthine-guanine phosphoribosyltransferase (HPRT) cause Lesch-Nyhan disease, a neurodevelopmental disorder characterized by cognitive, neurological, and behavioral abnormalities. Despite detailed knowledge of the enzyme's function, the key pathophysiological changes that accompany loss of purine recycling are unclear. To facilitate delineating the consequences of HPRT deficiency, four independent HPRT-deficient sublines of the human dopaminergic neuroblastoma, SK-N-BE(2) M17, were isolated by targeted mutagenesis with triple helix-forming oligonucleotides. As a group, these HPRT-deficient cells showed several significant abnormalities: (i) impaired purine recycling with accumulation of hypoxanthine, guanine, and xanthine, (ii) reduced guanylate energy charge and GTP : GDP ratio, but normal adenylate energy charge and no changes in any adenine nucleotide ratios, (iii) increased levels of UTP and NADP(+), (iv) reduced DOPA decarboxylase, but normal monoamines, and (v) reduction in cell soma size. These cells combine the analytical power of multiple lines and a human, neuronal origin to provide an important tool to investigate the pathophysiology of HPRT deficiency.