PDF(Pubmed)
Abstract：
Hematopoietic stem cell gene therapy (HSCGT) is a promising therapeutic strategy for the treatment of neurodegenerative, metabolic disorders. The approach involves the ex vivo introduction of a missing gene into patients\' own stem cells via lentiviral-mediated transduction (TD). Once transplanted back into a fully conditioned patient, these genetically modified HSCs can repopulate the blood system and produce the functional protein, previously absent or non-functional in the patient, which can then cross-correct other affected cells in somatic organs and the central nervous system. We previously developed an HSCGT approach for the treatment of Mucopolysaccharidosis type II (MPSII) (Hunter syndrome), a debilitating pediatric lysosomal disorder caused by mutations in the iduronate-2-sulphatase (IDS) gene, leading to the accumulation of heparan and dermatan sulfate, which causes severe neurodegeneration, skeletal abnormalities, and cardiorespiratory disease. In HSCGT proof-of-concept studies using lentiviral IDS fused to a brain-targeting peptide ApoEII (IDS.ApoEII), we were able to normalize brain pathology and behavior of MPSII mice. Here we present an optimized and validated good manufacturing practice hematopoietic stem cell TD protocol for MPSII in preparation for first-in-man studies. Inclusion of TEs LentiBOOST and protamine sulfate significantly improved TD efficiency by at least 3-fold without causing adverse toxicity, thereby reducing vector quantity required.
摘要：
造血干细胞基因治疗(HSCGT)是治疗神经退行性疾病的一种有前途的治疗策略,代谢紊乱。该方法涉及通过慢病毒介导的转导(TD)将缺失基因离体引入患者自身的干细胞。一旦移植回一个完全有条件的病人，这些基因修饰的HSC可以重新填充血液系统并产生功能性蛋白质，患者先前不存在或无功能，然后可以交叉校正体细胞器官和中枢神经系统中其他受影响的细胞。我们以前开发了一种用于治疗II型粘多糖贮积症（MPSII）（亨特综合征）的HSCGT方法，由艾杜糖醛酸-2-硫酸酯酶（IDS）基因突变引起的衰弱性小儿溶酶体疾病，导致乙酰肝素和硫酸皮肤素的积累，导致严重的神经变性，骨骼异常,和心肺疾病。在HSCGT概念验证研究中，使用与脑靶向肽ApoEII融合的慢病毒IDS（IDS。ApoEII),我们能够使MPSII小鼠的脑病理学和行为正常化。在这里，我们提出了一个优化和验证的良好生产实践造血干细胞TD协议为MPSII准备首次在人的研究。包含TEsLentiBOOST和硫酸鱼精蛋白可将TD效率显着提高至少3倍，而不会引起不良毒性，从而减少所需的矢量数量。
