Department of Internal Medicine, Washington University School of Medicine, 660 S. Euclid Ave., St Louis, MO 63110, USA.
Ann Neurol. 2012 Jun;71(6):797-804. doi: 10.1002/ana.23545. Epub 2012 Feb 24.
Infantile neuronal ceroid lipofuscinosis (INCL) is an inherited childhood neurodegenerative disorder caused by the loss of palmitoyl protein thioesterase-1 (PPT1) activity. Affected children suffer from blindness, epilepsy, motor dysfunction, cognitive decline, and premature death. The Ppt1(-/-) mouse shares the histological and clinical features of INCL. Previous single-therapy approaches using small molecule drugs, gene therapy, or neuronal stem cells resulted in partial histological correction, with minimal improvements in motor function or lifespan. Here, we combined central nervous system (CNS)-directed adeno-associated virus (AAV)2/5-mediated gene therapy with bone marrow transplantation (BMT) in the INCL mouse.
At birth, Ppt1(-/-) and wild-type mice were given either intracranial injections of AAV2/5-PPT1 or bone marrow transplantation, separately as well as in combination. To assess function, we measured rotorod performance monthly as well as lifespan. At terminal time points, we evaluated the therapeutic effects on several INCL-specific parameters, such as cortical thickness, autofluorescent accumulation, and glial activation. Finally, we determined levels of PPT1 enzyme activity and bone marrow engraftment in treated mice.
AAV2/5-mediated gene therapy alone resulted in significant histological correction, improved motor function, and increased lifespan. Interestingly, the addition of BMT further increased the lifespan of treated mice and led to dramatic, sustained improvements in motor function. These data are truly striking, given that BMT alone is ineffective, yet it synergizes with CNS-directed gene therapy to dramatically increase efficacy and lifespan.
AAV2/5-mediated gene therapy in combination with BMT provides an unprecedented increase in lifespan as well as dramatic improvement on functional and histological parameters.
婴儿神经元蜡样脂褐质沉积症（INCL）是一种遗传性儿童神经退行性疾病，由棕榈酰蛋白硫酯酶-1（PPT1）活性丧失引起。受影响的儿童患有失明、癫痫、运动功能障碍、认知能力下降和过早死亡。 Ppt1(-/-) 小鼠具有 INCL 的组织学和临床特征。以前使用小分子药物、基因治疗或神经元干细胞的单一治疗方法导致部分组织学纠正，运动功能或寿命仅有微小改善。在这里，我们将中枢神经系统 (CNS) 定向腺相关病毒 (AAV)2/5 介导的基因治疗与骨髓移植 (BMT) 相结合用于 INCL 小鼠。
出生时，将 Ppt1(-/-) 和野生型小鼠分别给予颅内注射 AAV2/5-PPT1 或骨髓移植，或两者结合。为了评估功能，我们每月测量转棒性能并记录寿命。在终末时间点，我们评估了皮质厚度、自发荧光积累和神经胶质激活等几个 INCL 特异性参数的治疗效果。最后，我们确定了治疗小鼠中 PPT1 酶活性和骨髓植入的水平。
AAV2/5 介导的基因治疗单独使用可导致显著的组织学纠正、运动功能改善和寿命延长。有趣的是，BMT 的加入进一步延长了治疗小鼠的寿命，并导致运动功能的显著、持续改善。鉴于 BMT 本身无效，但与 CNS 定向基因治疗协同作用可显著提高疗效和寿命，这些数据确实令人瞩目。
AAV2/5 介导的基因治疗与 BMT 相结合可显著延长寿命，并显著改善功能和组织学参数。
