Karumuthil-Melethil Subha, Marshall Michael S, Heindel Clifford, Jakubauskas Benas, Bongarzone Ernesto R, Gray Steven J
Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
Department of Anatomy and Cell Biology, University of Illinois at Chicago, Chicago, Illinois.
J Neurosci Res. 2016 Nov;94(11):1138-51. doi: 10.1002/jnr.23882.
Globoid cell leukodystrophy (GLD), or Krabbe disease, is an autosomal recessive neurodegenerative disease caused by the deficiency of the lysosomal enzyme galactocerebrosidase (GALC). Hematopoietic stem cell transplantation (HSCT) provides modest benefit in presymptomatic patients but is well short of a cure. Gene transfer experiments using viral vectors have shown some success in extending the survival in the mouse model of GLD, twitcher mice. The present study compares three single-stranded (ss) AAV serotypes, two natural and one engineered (with oligodendrocyte tropism), and a self-complementary (sc) AAV vector, all packaged with a codon-optimized murine GALC gene. The vectors were delivered via a lumbar intrathecal route for global CNS distribution on PND10-11 at a dose of 2 × 10(11) vector genomes (vg) per mouse. The results showed a similar significant extension of life span of the twitcher mice for all three serotypes (AAV9, AAVrh10, and AAV-Olig001) as well as the scAAV9 vector, compared to control cohorts. The rAAV gene transfer facilitated GALC biodistribution and detectable enzymatic activity throughout the CNS as well as in sciatic nerve and liver. When combined with BMT from syngeneic wild-type mice, there was significant improvement in survival for ssAAV9. Histopathological analysis of brain, spinal cord, and sciatic nerve showed significant improvement in preservation of myelin, with ssAAV9 providing the greatest benefit. In summary, we demonstrate that lumbar intrathecal delivery of rAAV/mGALCopt can significantly enhance the life span of twitcher mice treated at PND10-11 and that BMT synergizes with this treatment to improve the survival further. © 2016 Wiley Periodicals, Inc.
球形细胞脑白质营养不良（GLD），即克拉伯病，是一种常染色体隐性神经退行性疾病，由溶酶体酶半乳糖脑苷脂酶（GALC）缺乏引起。造血干细胞移植（HSCT）对症状前患者有一定益处，但远非治愈方法。使用病毒载体的基因转移实验在延长GLD小鼠模型（震颤小鼠）的生存期方面已取得一些成功。本研究比较了三种单链（ss）腺相关病毒血清型，两种天然血清型和一种经过改造具有少突胶质细胞嗜性的血清型，以及一种自我互补（sc）腺相关病毒载体，它们均包装有密码子优化的小鼠GALC基因。这些载体在出生后第10 - 11天通过腰椎鞘内途径递送，以实现中枢神经系统的整体分布，每只小鼠的剂量为2×10¹¹载体基因组（vg）。结果显示，与对照队列相比，所有三种血清型（AAV9、AAVrh10和AAV - Olig001）以及scAAV9载体都使震颤小鼠的寿命有相似的显著延长。重组腺相关病毒基因转移促进了GALC在整个中枢神经系统以及坐骨神经和肝脏中的生物分布和可检测的酶活性。当与同基因野生型小鼠的骨髓移植相结合时，ssAAV9的生存期有显著改善。对脑、脊髓和坐骨神经的组织病理学分析表明，髓磷脂的保存有显著改善，ssAAV9带来的益处最大。总之，我们证明在出生后第10 - 11天对震颤小鼠进行腰椎鞘内递送rAAV / mGALCopt可显著延长其寿命，并且骨髓移植与这种治疗协同作用可进一步提高生存率。© 2016威利期刊公司
