Biffi Alessandra, De Palma Michele, Quattrini Angelo, Del Carro Ubaldo, Amadio Stefano, Visigalli Ilaria, Sessa Maria, Fasano Stefania, Brambilla Riccardo, Marchesini Sergio, Bordignon Claudio, Naldini Luigi
San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scienctific Institute, Milan, Italy.
J Clin Invest. 2004 Apr;113(8):1118-29. doi: 10.1172/JCI19205.
Gene-based delivery can establish a sustained supply of therapeutic proteins within the nervous system. For diseases characterized by extensive CNS and peripheral nervous system (PNS) involvement, widespread distribution of the exogenous gene may be required, a challenge to in vivo gene transfer strategies. Here, using lentiviral vectors (LVs), we efficiently transduced hematopoietic stem cells (HSCs) ex vivo and evaluated the potential of their progeny to target therapeutic genes to the CNS and PNS of transplanted mice and correct a neurodegenerative disorder, metachromatic leukodystrophy (MLD). We proved extensive repopulation of CNS microglia and PNS endoneurial macrophages by transgene-expressing cells. Intriguingly, recruitment of these HSC-derived cells was faster and more robust in MLD mice. By transplanting HSCs transduced with the arylsulfatase A gene, we fully reconstituted enzyme activity in the hematopoietic system of MLD mice and prevented the development of motor conduction impairment, learning and coordination deficits, and neuropathological abnormalities typical of the disease. Remarkably, ex vivo gene therapy had a significantly higher therapeutic impact than WT HSC transplantation, indicating a critical role for enzyme overexpression in the HSC progeny. These results indicate that transplantation of LV-transduced autologous HSCs represents a potentially efficacious therapeutic strategy for MLD and possibly other neurodegenerative disorders.
基于基因的递送可在神经系统内建立治疗性蛋白质的持续供应。对于以中枢神经系统（CNS）和外周神经系统（PNS）广泛受累为特征的疾病，可能需要外源性基因的广泛分布，这对体内基因转移策略构成了挑战。在此，我们使用慢病毒载体（LV）在体外高效转导造血干细胞（HSC），并评估其后代将治疗性基因靶向移植小鼠的CNS和PNS以及纠正神经退行性疾病——异染性脑白质营养不良（MLD）的潜力。我们证明了转基因表达细胞可广泛重新填充CNS小胶质细胞和PNS神经内膜巨噬细胞。有趣的是，在MLD小鼠中，这些源自HSC的细胞的募集更快且更强劲。通过移植用芳基硫酸酯酶A基因转导的HSC，我们在MLD小鼠的造血系统中完全恢复了酶活性，并预防了运动传导障碍、学习和协调缺陷以及该疾病典型的神经病理异常的发展。值得注意的是，体外基因治疗比野生型HSC移植具有显著更高的治疗效果，表明酶在HSC后代中的过表达起关键作用。这些结果表明，LV转导的自体HSC移植代表了一种针对MLD以及可能其他神经退行性疾病的潜在有效治疗策略。
