Induction of neuronal differentiation of stem cell or cancer cell with VHL-gene transfer and neuronal regeneration using the transferred cells

通过野生型VHL(von Hippel-Lindau)基因、突变型VHL基因和反义VHL基因的转导，我们发现VHL蛋白具有诱导神经前体细胞和神经母细胞瘤细胞分化的能力。神经前体细胞可能为帕金森病(PD)的治疗提供多巴胺能神经元。然而，用目前的方法将神经干细胞移植到纹状体内的成功是有限的。在模型大鼠身上有可能逆转帕金森病的症状，但在人类身上很难逆转，因为神经前体细胞产生的多巴胺能神经元数量很少(不到1000个细胞)。我们将VHL基因导入从胎鼠脑中分离的神经干细胞中。携带VHL基因的神经干细胞在体外可有效分化为酪氨酸羟化酶(TH)阳性神经元。将预先用溴脱氧尿嘧啶核苷标记的携带VHL基因的神经干细胞移植到帕金森病模型大鼠纹状体。在移植部位附近可见大量的溴脱氧尿嘧啶核苷-TH双标细胞，表明移植细胞在宿主纹状体内有效地产生了新的多巴胺能神经元。此外，所有患有鼻咽癌并VHL的动物均显示阿朴吗啡诱导的旋转显著减少。这些发现表明，携带VHL基因的神经干细胞可以有效地产生多巴胺能神经元，并且足够数量的多巴胺能神经元(超过20,000个细胞)可以发展成逆转人类帕金森病症状的细胞。VHL基因转导为帕金森病的治疗提供了新的途径。

With transductions of wild-type VHL (von Hippel-Lindau) gene, mutated-type VHL gene, and anti-sense VHL gene, we showed VHL protein had induction ability of neuronal differentiation in neural progenitor cells and neuroblastoma cells. NPCs may provide dopaminergic neurons for the treatment of Parkinson's disease (PD). However, transplantation of NPCs into the striatum by current methods has had limited success. It is possible to reverse the symptoms of PD in model rats but difficult to reverse them in humans because the number of dopaminergic neurons generated from NPCs is small (less than 1000 cells). We transduced VHL gene into NPCs isolated from embryonic rat brain. The NPCs with the trunsduced VHL gene efficiently differentiated into tyrosine hydroxylase (TH)-positive neurons in vitro. NPCs with the trunsduced VHL gene, which were labeled in advance with bromodeoxyuridine, were transplanted into the striatum of a rat model of PD. Numerous bromodeoxyuridine-TH double-labeled cells were seen close to the transplant site, showing that the transplanted cells efficiently generated new dopaminergic neurons within he host striatum. Moreover, all of the animals with NPCs with VHL showed a remarkable decrease in apomorphine-induced rotations. These findings show that NPCs with the VHL gene can efficiently generate dopaminergic neurons and that sufficient a number of dopaminergic neurons (more than 20,000 cells) can develop from to reverse the symptoms of PD in humans. VHL gene transduction provides a new therapeutic approach for treatment of PD.