Mouse Models of Inherited Metabolic Disorders

法布里病是一种致命的X连锁隐性代谢性疾病， 由于溶酶体活性不足而引起的疾病 酶，α-半乳糖苷酶A（阿加）。在受影响的半合子男性中， 基质在血管溶酶体中进行性沉积 内皮细胞和平滑肌细胞导致闭塞性血管 疾病到目前为止，还没有针对这种情况的具体治疗方法。 酶替代和基因治疗都在 考虑，但在人体内进行这些试验将是困难的 而且耗时。我们已经开发了Fabry小鼠模型， 将是有价值的发展这样的治疗方案。这个鼠标 通过破坏小鼠中的阿加基因组位点来产生模型 胚胎干细胞同源重组虽然 这些小鼠完全缺乏阿加活性， 10周龄时临床正常。超微结构分析研究 显示肾脏中有同心层状包涵体， 使用荧光标记的凝集素进行显微镜检查， α-D-半乳糖残基显示底物在细胞中积累， 肾脏以及培养的成纤维细胞中。血脂分析显示 肝脏和肾脏中有明显的底物积累。 老化研究显示， 在80周龄的小鼠中具有亚临床肾脏病理学。骨 Fabry小鼠骨髓移植的实验研究 型小鼠纠正了大多数靶细胞的代谢缺陷， 表明其在临床领域的价值。我们早些时候 显示了阿加缺乏和积累的纠正 培养的阿加成纤维细胞中的α-D-半乳糖残基 使用含有人阿加的双顺反子MDR载体的无效小鼠 cDNA。这些研究扩展到体内校正， 脂质体作为递送载体。

Fabry disease is a fatal X-linked recessive metabolic disorder resulting from the deficient activity of the lysosomal enzyme, a-galactosidase A (AGA). In affected hemizygous males, the progressive deposition of substrate in lysosomes of vascular endothelial and smooth muscle cells causes occlusive vascular disease. To date, there is no specific treatment for this condition. Both enzyme replacement and gene therapy are under consideration, but carrying out these trials in human will be difficult and time-consuming. We have developed Fabry mouse model which will be valuable to develop such therapeutic regimes. This mouse model was generated by disrupting AGA genomic locus in mouse embryonic stem cells by homologous recombination. Although these mice showed a complete lack of AGA activity, they appeared clinically normal at 10 weeks of age. Ultrastructural analysis studies revealed concentric lamellar inclusions in the kidneys and confocal microscopy using a fluorescent-labeled lectin specific for a-D-galactosyl residues showed accumulation of substrate in the kidneys as well as in the cultured fibroblasts. Lipid analysis revealed a marked accumulation of the substrate in the liver and the kidneys. Aging studies revealed progressive accumulation of the substrate with sub-clinical kidney pathology in 80 weeks old mice. Bone marrow transplant of the Fabry mice with bone marrow from wild type mice corrected the metabolic defects in most of the target tissues indicating its value in the clinical domain.We had earlier shown correction of AGA deficiency and accumulation of a-D-galactosyl residues in the cultured fibroblasts from the AGA null mice using biscistronic MDR vectors containing human AGA cDNA. These studies were extended for in vivo correction using liposomes as delivery vehicles.