Li Wenzhong, Ma Nan, Ong Lee-Lee, Kaminski Alexander, Skrabal Christian, Ugurlucan Murat, Lorenz Peter, Gatzen Hans-Heinrich, Lützow Karola, Lendlein Andreas, Pützer Brigitte M, Li Ren-Ke, Steinhoff Gustav
Department of Cardiac Surgery, University of Rostock, Rostock, Germany.
J Gene Med. 2008 Aug;10(8):897-909. doi: 10.1002/jgm.1208.
Systemic gene delivery is limited by the adverse hydrodynamic conditions on the collection of gene carrier particles to the specific area. In the present study, a magnetic field was employed to guide magnetic nanobead (MNB)/polymer/DNA complexes after systemic administration to the left side of the mouse thorax in order to induce localized gene expression.
Nonviral polymer (poly ethyleneimine, PEI) vector-gene complexes were conjugated to MNBs with the Sulfo-NHS-LC-Biotin linker. In vitro transfection efficacy of MNB/PEI/DNA was compared with PEI/DNA in three different cell lines as well as primary endothelial cells under magnetic field stimulation. In vivo, MNB/PEI/DNA complexes were injected into the tail vein of mice and an epicardial magnet was employed to attract the circulating MNB/PEI/DNA complexes.
Endocytotic uptake of MNB/PEI/DNA complexes and intracellular gene release with nuclear translocation were observed in vitro, whereas the residues of MNB/PEI complexes were localized at the perinuclear region. Compared with PEI/DNA complexes alone, MNB/PEI/DNA complexes had a 36- to 85-fold higher transfection efficiency under the magnetic field. In vivo, the epicardial magnet effectively attracted MNB/PEI/DNA complexes in the left side of the thorax, resulting in strong reporter and therapeutic gene expression in the left lung and the heart. Gene expression in the heart was mainly within the endothelium.
MNB-mediated gene delivery could comprise a promising method for gene delivery to the lung and the heart.
全身基因递送受到基因载体颗粒在特定区域聚集时不利的流体动力学条件的限制。在本研究中，采用磁场引导全身给药后的磁性纳米珠（MNB）/聚合物/DNA复合物至小鼠胸部左侧，以诱导局部基因表达。
非病毒聚合物（聚乙烯亚胺，PEI）载体-基因复合物通过磺基-NHS-LC-生物素接头与MNB偶联。在磁场刺激下，比较了MNB/PEI/DNA与PEI/DNA在三种不同细胞系以及原代内皮细胞中的体外转染效率。在体内，将MNB/PEI/DNA复合物注入小鼠尾静脉，并使用心外膜磁体吸引循环中的MNB/PEI/DNA复合物。
在体外观察到MNB/PEI/DNA复合物的内吞摄取以及细胞内基因释放和核转位，而MNB/PEI复合物的残留物定位于核周区域。与单独的PEI/DNA复合物相比，MNB/PEI/DNA复合物在磁场下的转染效率高36至85倍。在体内，心外膜磁体有效地吸引了胸部左侧的MNB/PEI/DNA复合物，导致左肺和心脏中报告基因和治疗性基因的强烈表达。心脏中的基因表达主要在内皮细胞内。
MNB介导的基因递送可能是一种有前景的向肺和心脏递送基因的方法。
