Department of Physiology, Biophysics and Neurosciences, CINVESTAV, Mexico City, Mexico.
Department of Pharmacology, Wayne State University School of Medicine, Detroit, MI, USA.
Nanomedicine. 2017 May;13(4):1363-1375. doi: 10.1016/j.nano.2017.02.006. Epub 2017 Feb 17.
The human glial-cell derived neurotrophic factor (hGDNF) gene transfer by neurotensin (NTS)-polyplex nanoparticles functionally restores the dopamine nigrostriatal system in experimental Parkinson's disease models. However, high levels of sustained expression of GDNF eventually can cause harmful effects. Herein, we report an improved NTS-polyplex nanoparticle system that enables regulation of hGDNF expression within dopaminergic neurons. We constructed NTS-polyplex nanoparticles containing a single bifunctional plasmid that codes for the reverse tetracycline-controlled transactivator advanced (rtTA-Adv) under the control of NBRE3x promoter, and for hGDNF under the control of tetracycline-response element (TRE). Another bifunctional plasmid contained the enhanced green fluorescent protein (GFP) gene. Transient transfection experiments in N1E-115-Nurr1 cells showed that doxycycline (100 ng/mL) activates hGDNF and GFP expression. Doxycycline (5 mg/kg, i.p.) administration in rats activated hGDNF expression only in transfected dopaminergic neurons, whereas doxycycline withdrawal silenced transgene expression. Our results offer a specific doxycycline-regulated system suitable for nanomedicine-based treatment of Parkinson's disease.
神经降压素（NTS）-多聚物纳米粒的人胶质细胞源性神经营养因子（hGDNF）基因转染通过功能恢复实验性帕金森病模型中的多巴胺黑质纹状体系统。然而，高水平的持续 GDNF 表达最终可能会产生有害影响。在此，我们报告了一种改进的 NTS-多聚物纳米粒系统，该系统可调节多巴胺能神经元中的 hGDNF 表达。我们构建了包含单个双功能质粒的 NTS-多聚物纳米粒，该质粒在 NBRE3x 启动子的控制下编码反向四环素调控的转录激活物高级（rtTA-Adv），并在四环素反应元件（TRE）的控制下编码 hGDNF。另一个双功能质粒包含增强型绿色荧光蛋白（GFP）基因。在 N1E-115-Nurr1 细胞中的瞬时转染实验表明，强力霉素（100ng/ml）激活 hGDNF 和 GFP 表达。在大鼠中给予强力霉素（5mg/kg，腹腔内注射）仅激活转染的多巴胺能神经元中的 hGDNF 表达，而强力霉素撤药则沉默转基因表达。我们的结果提供了一种适合基于纳米医学治疗帕金森病的特定强力霉素调控系统。
