A DIFFERENTIAL EFFICIENCY OF ADENOVIRUS-MEDIATED IN-VIVO GENE-TRANSFER INTO SKELETAL-MUSCLE CELLS OF DIFFERENT MATURITY
A DIFFERENTIAL EFFICIENCY OF ADENOVIRUS-MEDIATED IN-VIVO GENE-TRANSFER INTO SKELETAL-MUSCLE CELLS OF DIFFERENT MATURITY
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DOI：
10.1093/hmg/3.4.579
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发表时间：
1994-04-01
影响因子：
3.5
通讯作者：
KARPATI, G
中科院分区：
文献类型：
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作者：
ACSADI, G;JANI, A;KARPATI, G
具有不同报告基因插入物的自主复制缺陷型5型人腺病毒（AV）重组体的高滴度（10（11）-10（12）pfu/ml）悬浮液（CMV-荧光素酶（Lux），CMV-β-半乳糖苷酶（Lac Z），将RSV-Lux和RSV-Lac Z）注射到1 - 5日龄（组1）或35 - 45日龄（组2）正常小鼠的完整四头肌中，以及再生成年小鼠肌肉（组3）和35日龄mdx肌肉（组4）。10天和2个月后定量报告基因的表达。在注射后10天，所有报告基因的表达是非常高的腹腔注射（组1）的肌肉。第2组肌肉中的转导明显较少，第3组肌肉中的基因表达明显优于第2组肌肉。在通常存在自发再生的成年mdx肌肉（第4组）中，结果与第3组动物相似。在第1组动物注射后2个月，RSV-Lux表达甚至高于注射后10天。第1、2、3和4组中包含AV内化受体的α（v）-整联蛋白分子的细胞表面密度与AV转导率呈正相关。我们的结论是，高滴度（10（10）pfu/ml或以上）的腺病毒载体能够有效地转导体内仅未成熟的肌肉细胞，但不能转导成熟的肌纤维，这似乎与未成熟的肌肉细胞中的AV内化受体的表面密度较高，而成熟的肌纤维中的AV内化受体的表面密度较低有关。本研究的结果对Duchenne型肌营养不良症（DMD）等骨骼肌疾病的基因治疗具有重要意义。
High titre (10(11) - 10(12) pfu/ml) suspensions of autonomously replication-defective type 5 human adenovirus (AV) recombinants with different reporter gene inserts (CMV-Luciferase (Lux), CMV-beta-galactosidase (Lac Z), RSV-Lux and RSV-Lac Z) were injected into intact quadriceps muscles of 1 - 5 day old (Group 1) or 35 - 45 day old (Group 2) normal mice, as well as regenerating adult mouse muscles (Group 3) and 35 day old mdx muscles (Group 4). The expression of the reporter genes was quantitated 10 days and 2 months later. At 10 days postinjection all reporter gene expression was very high in the neonatally injected (Group 1) muscles. In Group 2 muscles the transduction was markedly less. In Group 3 muscles the gene expression was significantly better than in the Group 2 muscles. In adult mdx muscles (Group 4) where spontaneous regeneration is usually present, the results were similar to those in Group 3 animals. At 2 months post-injection in Group 1 animals, the RSV-Lux expression was even higher than at 10 days postinjection. The cell surface density of alpha(v)-integrin-containing molecules including the internalization receptor for AV in Groups 1, 2, 3 and 4 showed a positive correlation with AV transducibility. We conclude that adenovirus vector in high titre (10(10) pfu/ml or above) is capable of efficiently transducing only immature muscle cells but not mature muscle fibers in vivo and this appears to correlate with a higher surface density of the available AV internalization receptor in immature muscle cells and lower level in mature muscle fibers. The results of this study have important relevance to gene therapy of skeletal muscles in such diseases as Duchenne muscular dystrophy (DMD).