Mouse Models of Inherited Metabolic Disorders

法布里病是一种致命性X连锁隐性代谢 溶酶体活性不足引起的疾病 酶，α-半乳糖苷酶A(AGA)。在受影响的半合子男性中， 血管溶酶体中底物的进行性沉积 血管内皮细胞和平滑肌细胞导致血管闭塞 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进行中 考虑，但在人体上进行这些试验将是困难的 而且很耗时。我们已经开发了法布里老鼠模型，它可以 对开发这样的治疗方案将是有价值的。这只鼠标 破坏小鼠AGA基因座建立模型 胚胎干细胞通过同源重组。虽然 这些小鼠表现出完全缺乏AGA活性，他们似乎 10周龄时临床正常。超微结构分析研究 肾脏和共焦内可见同心的板层状包涵体 使用荧光标记的凝集素进行显微镜检查 A-D-半乳糖基残基显示底物在 在肾脏和培养的成纤维细胞中也是如此。血脂分析显示 肝脏和肾脏中底物的显著堆积。 老化研究显示底物逐渐积累 80周龄小鼠肾脏亚临床病理改变。骨 用野生型骨髓移植Fabry小鼠的研究 型小鼠纠正了大多数靶点的代谢缺陷 表明它在临床领域的价值的组织。我们之前 AGA缺乏和积聚的AGA显示纠正 AGA培养成纤维细胞中A-D-半乳糖残基的研究 携带人AGA双顺反子多药耐药载体的空白小鼠 CDNA.这些研究被扩展到活体矫正使用 脂质体作为输送载体。

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.