Dystrophin expression in myofibers of Duchenne muscular dystrophy patients following intramuscular injections of normal myogenic cells
Dystrophin expression in myofibers of Duchenne muscular dystrophy patients following intramuscular injections of normal myogenic cells
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DOI：
10.1016/j.ymthe.2003.11.023
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发表时间：
2004-03-01
影响因子：
12.4
通讯作者：
Tremblay, JP
中科院分区：
文献类型：
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作者：
Skuk, D;Roy, B;Tremblay, JP
三名Duchenne肌营养不良症(DMD)患者接受了从正常捐赠者的骨骼肌活检中获得的肌源性细胞的注射。将细胞(30×10(6))按25次平行注射的方式注入胫骨前1 cm内。作为对照，我们在对侧肌肉中进行了类似的生理盐水注射。患者接受他克莫司免疫抑制治疗。4周后在注射部位进行肌肉活组织检查。我们观察到肌营养不良蛋白阳性的肌纤维；在细胞移植部位，分别有9例(1例)、6.8例(2例)和11%(3例)。由于患者1和2已经确定了dystrophin基因的缺失，这些结果是使用针对缺失外显子编码的表位的单抗获得的。供体Dystrophin在对照部位缺失。患者3有外显子重复，因此特异性供体-肌营养不良蛋白检测是不可能的。然而，移植细胞中的抗肌营养不良蛋白阳性肌纤维比对照部位多四倍。患者1和2的供体-肌营养不良蛋白转录本仅在患者1和2的细胞移植部位被检测到(使用与缺失外显子的序列反应的引物)。在患者3的细胞移植部位，肌营养不良蛋白的转录本比对照部位更丰富。因此，在特定的细胞输送和免疫抑制条件下，DMD患者的骨骼肌中可以获得显著的肌营养不良蛋白表达。
Three Duchenne muscular dystrophy (DMD) patients received injections of myogenic cells obtained from skeletal muscle biopsies of normal donors. The cells (30 x 10(6)) were injected in 1 cm 3 of the tibialis anterior by 25 parallel injections. We performed similar patterns of saline injections in the contralateral muscles as controls. The patients received tacrolimus for immunosuppression. Muscle biopsies were performed at the injected sites 4 weeks later. We observed dystrophin-positive myofibers; in the cell-grafted sites amounting to 9 (patient 1), 6.8 (patient 2), and 11% (patient 3). Since patients I and 2 had identified dystrophin-gene deletions these results were obtained using monoclonal antibodies specific to epitopes coded by the deleted exons. Donor dystrophin was absent in the control sites. Patient 3 had exon duplication and thus specific donor-dystrophin detection was not possible. However, there were fourfold more dystrophin-positive myofibers in the cell-grafted than in the control site. Donor-dystrophin transcripts were detected by RT-PCR (using primers reacting with a sequence in the deleted exons) only in the cell-grafted sites in patients 1 and 2. Dystrophin transcripts were more abundant in the cell-grafted than in the control site in patient 3. Therefore, significant dystrophin expression can be obtained in the skeletal muscles of DMD patients following specific conditions of cell delivery and immunosuppression.