Division of Regenerative Medicine, Stem Cells and Gene Therapy, San Raffaele Scientific Institute, 20132 Milan, Italy.
Sci Transl Med. 2011 Aug 17;3(96):96ra78. doi: 10.1126/scitranslmed.3002342.
In contrast to conventional gene therapy vectors, human artificial chromosomes (HACs) are episomal vectors that can carry large regions of the genome containing regulatory elements. So far, HACs have not been used as vectors in gene therapy for treating genetic disorders. Here, we report the amelioration of the dystrophic phenotype in the mdx mouse model of Duchenne muscular dystrophy (DMD) using a combination of HAC-mediated gene replacement and transplantation with blood vessel-associated stem cells (mesoangioblasts). We first genetically corrected mesoangioblasts from dystrophic mdx mice with a HAC vector containing the entire (2.4 Mb) human dystrophin genetic locus. Genetically corrected mesoangioblasts engrafted robustly and gave rise to many dystrophin-positive muscle fibers and muscle satellite cells in dystrophic mice, leading to morphological and functional amelioration of the phenotype that lasted for up to 8 months after transplantation. Thus, HAC-mediated gene transfer shows efficacy in a preclinical model of DMD and offers potential for future clinical translation.
与传统的基因治疗载体相反，人类人工染色体（HACs）是能够携带含有调节元件的基因组大片段的附加体载体。到目前为止，HAC 尚未被用作治疗遗传疾病的基因治疗载体。在这里，我们报告了使用 HAC 介导的基因替换和与血管相关的干细胞（间充质成肌细胞）移植的组合，改善了 Duchenne 肌营养不良症（DMD）的 mdx 小鼠模型的营养不良表型。我们首先使用包含整个（2.4 Mb）人类肌营养不良蛋白遗传基因座的 HAC 载体对来自营养不良 mdx 小鼠的间充质成肌细胞进行基因校正。经过基因校正的间充质成肌细胞在营养不良的小鼠中大量植入，并产生了许多肌营养不良蛋白阳性的肌纤维和肌肉卫星细胞，导致表型的形态和功能得到改善，移植后持续了长达 8 个月。因此，HAC 介导的基因转移在 DMD 的临床前模型中显示出疗效，并为未来的临床转化提供了潜力。
