School of Clinical and Experimental Medicine, University of Birmingham, Edgbaston, UK.
Brain Res. 2010 Nov 4;1359:1-13. doi: 10.1016/j.brainres.2010.08.044. Epub 2010 Aug 21.
Because the choroid plexus normally controls the production and composition of cerebrospinal fluid and, as such, its many functions of the central nervous system, we investigated whether ligand-mediated targeting could deliver genes to its secretory epithelium. We show here that when bacteriophages are targeted with epidermal growth factor, they acquire the ability to enter choroid epithelial cells grown in vitro as cell cultures, ex vivo as tissue explants or in vivo by intracerebroventricular injection. The binding and internalization of these particles activate EGF receptors on targeted cells, and the dose- and time-dependent internalization of particles is inhibited by the presence of excess ligand. When the phage genome is further reengineered to contain like green fluorescent protein or firefly luciferase under control of the cytomegalovirus promoter, gene expression is detectable in the choroid plexus and ependymal epithelium by immunohistochemistry or by noninvasive imaging, respectively. Taken together, these data support the hypothesis that reengineered ligand-mediated gene delivery should be considered a viable strategy to increase the specificity of gene delivery to the central nervous system and bypass the blood-brain barrier so as to exploit the biological effectiveness of the choroid plexus as a portal of entry into the brain.
由于脉络丛通常控制着脑脊液的产生和组成，以及中枢神经系统的许多功能，我们研究了配体介导的靶向是否可以将基因递送到其分泌上皮。我们在这里表明，当噬菌体被表皮生长因子靶向时，它们获得了进入体外培养的脉络丛上皮细胞的能力，无论是作为细胞培养物、组织外植体还是通过脑室内注射在体内。这些颗粒的结合和内化激活了靶向细胞上的表皮生长因子受体，并且存在过量配体时会抑制颗粒的剂量和时间依赖性内化。当噬菌体基因组进一步被重新设计为在巨细胞病毒启动子的控制下包含绿色荧光蛋白或荧光素酶时，通过免疫组织化学或非侵入性成像分别可以在脉络丛和室管膜上皮中检测到基因表达。总之，这些数据支持这样一种假设，即经过重新设计的配体介导的基因传递应该被认为是一种可行的策略，以增加基因传递到中枢神经系统的特异性，并绕过血脑屏障，从而利用脉络丛作为进入大脑的门户的生物学有效性。
