Polybutylcyanoacrylate Nanoparticle-Mediated Neurotrophin-3 Gene Delivery for Treating Intracerebral Hemorrhage

• Main Authors: Chiu-Yen Chung, 鐘秋燕
• Other Authors: Yung-Chih Kuo
• Format: Others
• Language: en_US
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/81165078196008436976

博士 === 國立中正大學 === 化學工程研究所 === 101 === The aim of this study was to show the capability of a non-viral vector-mediated gene delivery system comprising Polybutylcyanoacrylate (PBCA) nanoparticles (NPs) and cmvNT-3-HRE in treating hypertensive intracerebral hemorrhage (ICH) rats. In the first part of studies, we went to identify the functional of gene construct, PBCA NP-mediated neurotrophin-3 (NT-3) gene delivery for differentiating induced pluripotent stem cell (iPSCs) into neuron. The data indicate that PBCA NPs could efficiently grasp NT-3 gene, thereby increase the particle size and confer a negative surface charge. In addition, the treatments with PBCA NP/NT-3 complexes enhanced the expression of NT-3, TrkC, NH-H, NSE, and PSD95 by differentiating iPSCs. In conclusion, PBCA NPs can serve as an efficient gene delivery vector in enhancing transfection of iPSCs with cmvNT-3 to prompt their differentiation toward neurons. In the second part of studies, the rats subjected to the collagenase-induced ICH were used to mimic hypertensive intracerebral hemorrhage in human. We construct a NT-3 expression plasmid containing the hypoxia-responsive element (HRE) and a cytomegalovirus (cmv) promotor. PBCA NP-NT-3 plasmid containing HRE is applied to activate the hypoxia inducible factor-1 (HIF-1)/HRE system of gene regulation and used to produce NT-3 proteins after ICH. The data indicate that the PBCA NPs/cmvNT-3-HRE complexes are very low toxicity. Treatments with PBCA NP/cmvNT-3-HRE complexes increased the expression of NT-3, TrkC, and MAP-2 by differentiating iPSCs under hypoxia condition. Furthermore, the PBCA NPs can protect cmvNT-3-HRE from degradation in vitro, sustain across the blood-brain barrier (BBB) in vivo, enhance the NT-3 protein expression, inhibit the AIF, cleavaged caspase-3 and DNA fragmentation expression, and increase neuros survival after hemorrhagic stroke. In conclusion, the PBCA NPs are justified as an appropriate system for gene delivery in the brain because of it possibly influences therapeutic outcome of ICH rats.