Division of Human Genetics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
Hum Mol Genet. 2019 Oct 15;28(20):3406-3421. doi: 10.1093/hmg/ddz184.
Gaucher disease (GD) is caused by GBA1 mutations leading to functional deficiency of acid-β-glucosidase (GCase). No effective treatment is available for neuronopathic GD (nGD). A subclass of neural stem and precursor cells (NPCs) expresses VLA4 (integrin α4β1, very late antigen-4) that facilitates NPC entry into the brain following intravenous (IV) infusion. Here, the therapeutic potential of IV VLA4+NPCs was assessed for nGD using wild-type mouse green fluorescent protein (GFP)-positive multipotent induced pluripotent stem cell (iPSC)-derived VLA4+NPCs. VLA4+NPCs successfully engrafted in the nGD (4L;C*) mouse brain. GFP-positive cells differentiated into neurons, astrocytes and oligodendrocytes in the brainstem, midbrain and thalamus of the transplanted mice and significantly improved sensorimotor function and prolonged life span compared to vehicle-treated 4L;C* mice. VLA4+NPC transplantation significantly decreased levels of CD68 and glial fibrillary acidic protein, as well as TNFα mRNA levels in the brain, indicating reduced neuroinflammation. Furthermore, decreased Fluoro-Jade C and NeuroSilver staining suggested inhibition of neurodegeneration. VLA4+NPC-engrafted 4L;C* midbrains showed 35% increased GCase activity, reduced substrate [glucosylceramide (GC, -34%) and glucosylsphingosine (GS, -11%)] levels and improved mitochondrial oxygen consumption rates in comparison to vehicle-4L;C* mice. VLA4+NPC engraftment in 4L;C* brain also led to enhanced expression of neurotrophic factors that have roles in neuronal survival and the promotion of neurogenesis. This study provides evidence that iPSC-derived NPC transplantation has efficacy in an nGD mouse model and provides proof of concept for autologous NPC therapy in nGD.
戈谢病（GD）是由 GBA1 突变导致酸性-β-葡萄糖苷酶（GCase）功能缺陷引起的。目前还没有有效的治疗神经元病变型 GD（nGD）的方法。一类神经干细胞和前体细胞（NPCs）表达 VLA4（整合素 α4β1，即非常晚期抗原-4），这种蛋白能够促进 NPC 经静脉（IV）输注进入大脑。在这里，使用野生型小鼠绿色荧光蛋白（GFP）阳性多能诱导多能干细胞（iPSC）衍生的 VLA4+NPC 评估了 IV VLA4+NPC 治疗 nGD 的潜力。VLA4+NPC 成功地植入 nGD（4L;C*）小鼠的大脑中。移植小鼠的脑桥、中脑和丘脑中有 GFP 阳性细胞分化为神经元、星形胶质细胞和少突胶质细胞，与 vehicle 治疗的 4L;C小鼠相比，它们显著改善了感觉运动功能并延长了寿命。VLA4+NPC 移植显著降低了大脑中 CD68 和神经胶质纤维酸性蛋白的水平，以及 TNFα mRNA 水平，表明神经炎症减少。此外，减少的 Fluoro-Jade C 和 NeuroSilver 染色表明神经退行性变得到抑制。与 vehicle-4L;C小鼠相比，VLA4+NPC 移植的 4L;C中脑显示 GCase 活性增加 35%，底物[葡萄糖脑苷脂（GC，-34%）和葡萄糖鞘氨醇（GS，-11%）]水平降低，线粒体耗氧量增加。VLA4+NPC 移植到 4L;C大脑还导致神经营养因子表达增强，这些因子在神经元存活和促进神经发生中发挥作用。这项研究为 iPSC 衍生 NPC 移植在 nGD 小鼠模型中的疗效提供了证据，并为 nGD 的自体 NPC 治疗提供了概念验证。
