公开(公告)号：US06262035B1

公开(公告)日：2001-07-17

申请号：US09164664

申请日：1998-10-01

申请人： Kevin P. Campbell ,  Kathleen H. Holt ,  Franck Duclos ,  Leland E. Lim ,  Volker Straub ,  Beverly Davidson ,  Roger Williamson

发明人： Kevin P. Campbell ,  Kathleen H. Holt ,  Franck Duclos ,  Leland E. Lim ,  Volker Straub ,  Beverly Davidson ,  Roger Williamson

IPC分类号： A01N4304

CPC分类号： C12N15/8509 ,  A01K67/0276 ,  A01K2207/15 ,  A01K2217/00 ,  A01K2217/075 ,  A01K2227/105 ,  A01K2267/0306 ,  A61K38/1709 ,  A61K48/00

摘要： Disclosed is a method for treating a patient suffering from the disease sarcoglycan-deficient limb-girdle muscular dystrophy by gene replacement therapy. Sarcoglycan gene replacement therapy produces extensive long-term expression of the sarcoglycan species which restores the entire sarcoglycan complex, results in the stable association of alph&agr;-dystroglycan with the sarcolemma, and eliminates the morphological markers of limb-girdle muscular dystrophy. In another aspect, the invention relates to a method for determining a specific defective sarcoglycan species in the tissue of a patient. The method involves culture of muscle cells obtained from the patient, and the independent introduction of expression vectors encoding each of the sarcoglycan species, &agr;, &bgr;, &ggr;, and &dgr;, into the cultured cells with subsequent assaying for restoration of the dystrophin-glycoprotein complex. In another aspect, the invention relates to a mouse, and cells derived therefrom, homozygous for a disrupted &agr;-sarcoglycan gene. The disruption prevents the synthesis of functional &agr;-sarcoglycan in cells of the mouse and results in the mutant mouse having no detectable sarcospan, &bgr;-, &ggr;-, &dgr;-sarcoglycan, and reduced &agr;-dystroglycan in the sarcolemma of skeletal and cardiac muscles, and a reduction of dystrophin in skeletal muscle, when compared to tissue of a mouse lacking a disrupted &agr;-sarcoglycan gene. In another aspect, the invention relates to methods for screening for therapeutic agents useful in the treatment of sarcoglycan-deficient limb-girdle muscular dystrophy. The methods involve administering a candidate therapeutic agent to a mouse, or cells derived therefrom, and assaying for therapeutic effects on the mouse or cells, with the determination of therapeutic effects being a reduction or reversal in disease progression, or a restoration of the dystroglycan complex.

摘要翻译： 本发明公开了一种通过基因替代疗法治疗患有疾病肌糖肌苷缺乏症的肢体肌营养不良症的患者的方法。 Sarcoglycan基因替代疗法产生广泛的长期表达的sarcoglycan物种，其恢复整个slegoglycan复合物，导致αphorpha-dystroglycan与肌膜的稳定关联，并消除肢体肌营养不良的形态学标志物。 另一方面，本发明涉及一种用于确定患者组织中的特定缺陷型糖团聚物物种的方法。 该方法包括从患者获得的肌肉细胞的培养，以及将编码每种肌糖原聚糖物质，α，β，γ和δ的表达载体独立引入培养的细胞中，随后测定肌营养不良蛋白 - 糖蛋白复合物的恢复 。 另一方面，本发明涉及一种小鼠及其衍生的细胞，用于破坏的α-丝裂霉素基因是纯合的。 该破坏阻止了小鼠细胞中功能性α-sarcoglycan的合成，并导致突变小鼠在骨骼肌和心脏肌肉的肌膜中不具有可检测的sarcospan，β - ，γ-，δ-色氨酸聚糖和还原的α-二糖聚糖， 和与缺乏α-sarcoglycan基因的小鼠的组织相比，骨骼肌中营养不良蛋白的减少。 另一方面，本发明涉及筛选用于治疗肌糖原缺乏型肢体肌营养不良症的治疗剂的方法。 所述方法包括将候选治疗剂施用于小鼠或由其衍生的细胞，以及测定对小鼠或细胞的治疗效果，其中治疗效果的确定是疾病进展中的减少或逆转，或者肌萎缩糖复合物的恢复 。