Cell and Gene Therapy Center, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, No. 88 Keling Road, Suzhou New District, Suzhou, 215163, People's Republic of China.
CAS Key Lab of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, No. 88 Keling Road, Suzhou New District, Suzhou, 215163, People's Republic of China.
Neurochem Res. 2018 Apr;43(4):930-937. doi: 10.1007/s11064-018-2498-7. Epub 2018 Feb 12.
The neural stem cells (NSCs) have the ability to self-renew, and to migrate to pathologically altered regions of the central nervous system. Glial cell derived neurotrophic factor (GDNF) could protect dopamine neurons and rescue motor neurons in vivo, which has been proposed as a promising candidate for the treatments of degenerative neurological diseases. In order to combine the advantages of neurotrophic factors and stem cells in clinical therapy, we established the modified hNSCs that has site-specific integration of GDNF gene by using recombinant adeno-associated virus (rAAV) vectors. The hNSCs were co-infected by rAAV2-EGFP-GDNF and rAAV2-SVAV2 which provide integrase to specifically integrate GDNF gene into AAVS1 site. The GDNF-hNSCs maintained their original stem cell characteristics and the ability to differentiate into neurons in vitro. In the animal model, the GDNF-hNSCs were specifically transplanted into CA1 area of hippocampi and could migrate to the dentate gyrus region and differentiate into neuronal cells while maintaining GDNF expression. hNSCs with GDNF gene site-specific integration at AAVS1 by using AAV vectors retained their stemness and effectively expressed GDNF, which indicates the potential of employing transplanted hNPCs for treatment of brain injuries and degenerative neurological diseases.
神经干细胞（NSCs）具有自我更新和迁移到中枢神经系统病理性改变区域的能力。胶质细胞源性神经营养因子（GDNF）可以保护多巴胺神经元并挽救体内运动神经元，这使其成为治疗退行性神经疾病的有前途的候选药物。为了将神经营养因子和干细胞的优势结合到临床治疗中，我们通过使用重组腺相关病毒（rAAV）载体建立了修饰的 hNSC，其 GDNF 基因具有特定的整合位点。hNSC 被 rAAV2-EGFP-GDNF 和 rAAV2-SVAV2 共感染，rAAV2-SVAV2 提供整合酶，将 GDNF 基因特异性整合到 AAVS1 位点。GDNF-hNSC 保持其原始干细胞特性和体外分化为神经元的能力。在动物模型中，GDNF-hNSC 被特异性移植到海马 CA1 区，能够迁移到齿状回区并分化为神经元细胞，同时保持 GDNF 表达。使用 AAV 载体将 GDNF 基因特异性整合到 AAVS1 的 hNSC 保留了其干细胞特性并有效表达 GDNF，这表明移植的 hNPC 用于治疗脑损伤和退行性神经疾病的潜力。
