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

通过VHL基因转移诱导干细胞或癌细胞的神经元分化以及使用转移的细胞进行神经元再生

基本信息

批准号：
13557120
负责人：
KANNO Hiroshi
金额：
$ 3.9万
依托单位：
Yokohama City University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

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.

通过对野生型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蛋白在神经祖细胞和神经母细胞瘤细胞中具有诱导神经元分化的能力。npc可能为帕金森病（PD）的治疗提供多巴胺能神经元。然而，通过目前的方法将npc移植到纹状体的成功有限。在模型大鼠中有可能逆转PD的症状，但在人类中很难逆转，因为npc产生的多巴胺能神经元数量很少（少于1000个细胞）。我们将VHL基因转染到胚胎大鼠脑分离的npc中。转染VHL基因的神经细胞在体外高效分化为酪氨酸羟化酶（TH）阳性神经元。将诱导VHL基因的NPCs，事先用溴脱氧尿苷标记，移植到PD模型大鼠纹状体中。移植部位附近可见大量溴脱氧尿嘧啶- th双标记细胞，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NPCs伴VHL的动物都表现出阿吗啡诱导的旋转显著减少。这些发现表明，具有VHL基因的npc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）可以从PD发育到逆转人类PD症状。VHL基因转导为PD的治疗提供了新的途径。