GENE-TRANSFER INTO SKELETAL-MUSCLES BY ISOGENIC MYOBLASTS
GENE-TRANSFER INTO SKELETAL-MUSCLES BY ISOGENIC MYOBLASTS
复制标题
DOI：
10.1089/hum.1994.5.8-949
复制
发表时间：
1994-08-01
影响因子：
4.2
通讯作者：
KARPATI, G
中科院分区：
文献类型：
--
作者：
HUARD, J;ACSADI, G;KARPATI, G
克服异源成肌细胞移植（HMT）中免疫排斥的最佳方法是使用免疫缺陷和/或高度免疫抑制的小鼠作为宿主。这同样可以通过自体成肌细胞移植（AMT）来实现。在本文中，我们描述了成肌细胞转移mdx和正常小鼠的供体肌源性细胞来源于高度近亲繁殖的同窝被认为是同基因的，因此该程序是类似于AMT。通过自主复制缺陷型重组人腺病毒介导的转导，用劳斯肉瘤病毒（RSV）-荧光素酶（Lux）或RSV-β-半乳糖苷酶（LacZ）报告基因在体外标记成肌细胞。这使我们能够跟踪他们移植后的命运。mdx和正常小鼠用20 Gray γ射线照射;在成肌细胞注射之前通过肌内notexin诱导坏死和再生。在mdx和正常小鼠中，荧光素酶的表达在注射后迅速下降，这意味着由于未确定的原因，大部分注射的成肌细胞在48小时前丢失。存活的注射成肌细胞很好地镶嵌了大量的宿主纤维，但仅在注射附近。在正常小鼠中，报告基因的大量表达持续到移植后1个月，但在mdx小鼠中报告基因表达逐渐下降并最终消失。后一种现象是由于mdx中的肌纤维持续强烈坏死所致。没有免疫排斥的证据。这些实验表明，即使在没有免疫排斥的情况下，成肌细胞转移也具有重要的负面特征：在注射后48小时内成肌细胞大量损失，并且注射的细胞在宿主肌肉中从注射部位没有显著扩散。这些因素，加上杜氏肌营养不良症（DMD）成肌细胞有限的增殖和融合能力，使它们不是DMD中抗肌萎缩蛋白基因替代疗法的抗肌萎缩蛋白cDNA的理想载体。
The best way to overcome immunorejection in heterologous myoblast transfer (HMT) is by the use of immunodeficient and/or highly immunosuppressed mice as hosts. The same may be attained by autologous myoblast transfer (AMT). In this paper, we describe myoblast transfer in mdx and normal mice where the donor myogenic cells originated from highly inbred litter mates that are considered to be isogenic and thus the procedure is analogous to AMT. The myoblasts were marked in vitro with Rous Sarcoma Virus (RSV)-luciferase (Lux) or RSV-beta-galactosidase (LacZ) reporter genes through transduction mediated by an autonomously replication-defective recombinant human adenovirus. This permitted us to follow their fate after transplantation. mdx and normal mice were irradiated with 20 Gray gamma rays; necrosis and regeneration were induced by intramuscular notexin prior to myoblast injection. In both mdx and normal mice, the expression of luciferase rapidly declined after the injection implying that a large portion of the injected myoblasts were lost by 48 hr, due to undetermined cause(s). The surviving, injected myoblasts well-mosaicized large groups of host fibers but only in the immediate vicinity of the injection. Substantial expression of the reporter gene continued up to 1 month post-transplantation in normal mice, but there was a gradual decline and eventual disappearance of the reporter gene expression in mdx mice. This latter phenomenon was due to the ongoing intense necrosis of muscle fibers in mdx. There was no evidence of immunorejection. These experiments indicate that even in the absence of immunorejection, myoblast transfer suffers from important negative features: major loss of myoblasts within 48 hr after the injection and lack of significant spread of the injected cells from the injection site in the host muscle, These factors, plus the limited proliferative and fusion capacity of Duchenne muscular dystrophy (DMD) myoblasts, make them less than an ideal vector for the dystrophin cDNA for dystrophin gene replacement therapy in DMD.