In Vivo fusion of circulating fluorescent cells with dystrophin-deficient myofibers results in extensive sarcoplasmic fluorescence expression but limited dystrophin sarcolemmal expression
In Vivo fusion of circulating fluorescent cells with dystrophin-deficient myofibers results in extensive sarcoplasmic fluorescence expression but limited dystrophin sarcolemmal expression
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DOI：
10.1016/s0002-9440(10)62484-4
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发表时间：
2005-06-01
影响因子：
6
通讯作者：
Gherardi, RK
中科院分区：
文献类型：
--
作者：
Chretien, F;Dreyfus, PA;Gherardi, RK
为了研究骨髓移植对杜氏肌营养不良症的治疗潜力，将绿色荧光蛋白阳性 (GFP(+)) 骨髓细胞移植到受辐射的野生型和肌营养不良蛋白缺陷型 mdx 小鼠中。胫骨前肌的 mdx 中 GFP(+) 单核细胞数量是野生型小鼠的五到六倍，GFP(+) 肌纤维数量是野生型小鼠的三到四倍。相比之下，mdx 小鼠中的肌营养不良蛋白表达仍保持在未移植 mdx 小鼠的水平内，并且与 GFP 共表达的情况很少见。 5000 根肌纤维的纵向切片显示 160 根 GFP+ 纤维，其中 9 根共表达肌营养不良蛋白。 GFP 始终被可视化为超过样本长度跨度（高达 1500 μm）的全长肌浆荧光，而肌营养不良蛋白表达仅限于该长度的 11% 至 28%。 GFP(+)纤维中的抗肌营养不良蛋白表达跨度(116+/-46μm)比回复纤维(654+/-409μm)短得多。这些数据表明，可溶性 GFP 远离与预先存在的肌营养不良蛋白 (-) 肌纤维的融合位点扩散，而肌营养不良蛋白仍然主要在融合位点附近表达。由于需要恢复整个肌纤维长度中的抗肌营养不良蛋白才能预期杜氏肌营养不良症的功能改善和临床益处，因此细胞疗法在神经肌肉疾病中的未来应用可以更合适地设想用于替代有缺陷的可溶性肌浆蛋白。
To investigate the therapeutic potential of bone marrow transplantation in Duchenne muscular dystrophy, green fluorescent protein-positive (GFP(+)) bone marrow cells were transplanted into irradiated wildtype and dystrophin-deficient mdx mice. Tibialis anterior muscles showed fivefold to sixfold more GFP(+) mononucleated cells and threefold to fourfold more GFP(+) myofibers in mdx than in wild-type mice. In contrast, dystrophin expression in mdx mice remained within the level of nontransplanted mdx mice, and co-expression with GFP was rare. Longitudinal sections of 5000 myofibers showed 160 GFP+ fibers, including 9 that co-expressed dystrophin. GFP was always visualized as full-length sarcoplasmic fluorescence that exceeded the span of sample length (up to 1500 mu m), whereas dystrophin expression was restricted to 11 to 28 % of this length. Dystrophin expression span was much shorter in GFP(+) fibers (116 +/- 46 mu m) than in revertant fibers (654 +/- 409 mu m). These data suggest that soluble GFP diffuses far from the fusion site with a pre-existing dystrophin(-) myofiber whereas dystrophin remains mainly expressed close to the site of fusion. Because restoration of dystrophin in whole muscle fiber length is required to expect functional improvement and clinical benefits for Duchenne muscular dystrophy, future applications of cell therapies to neuromuscular disorders could be more appropriately envisaged for replacement of defective soluble sarcoplasmic proteins.