The ependymal route for insulin-like growth factor-1 gene therapy in the brain.
The ependymal route for insulin-like growth factor-1 gene therapy in the brain.
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DOI：
10.1016/j.neuroscience.2009.06.024
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发表时间：
2009-09-29
期刊：
影响因子：
3.3
通讯作者：
Goya, R. G.
中科院分区：
文献类型：
--
作者：
Herenu, C. B.;Sonntag, W. E.;Morel, G. R.;Portiansky, E. L.;Goya, R. G.
脑室内施用肽胰岛素样生长因子-1（IGF-1）已被证明是不同动物模型脑中的有效神经保护策略，主要优点是在脑中实现高浓度的IGF-1而不改变肽的血清水平。为了进一步利用这种治疗方法，我们使用高效重组腺病毒（RAd）载体表达其转基因的控制下，有效的小鼠巨细胞病毒立即早期（mCMV）启动子，以高效率地抑制脑室管膜细胞，并实现有效释放转基因IGF-1到脑脊液（CSF）。我们构建了表达嵌合蛋白（TK/GFP）fus（与HSV 1胸苷激酶融合的绿色荧光蛋白）或编码大鼠IGF-1的cDNA的RAd载体，两者均由mCMV启动子驱动。将载体注射到年轻大鼠的侧脑室中，并通过荧光显微镜评估脑切片中嵌合GFP的表达。室管膜细胞标记波形蛋白通过免疫荧光检测，细胞核用DNA染料DAPI标记。在载体注射后的不同时间抽取血液和CSF样品。在所有的脑室，波形蛋白免疫反应细胞的室管膜主要转导的RAd-（TK/GFP）融合蛋白，显示核和胞质表达的转基因。对于伸长细胞（TK/GFP），fus表达在其细胞质过程中是明显的，因为它们深入到下丘脑实质。侧脑室注射RAd-IGF-1可诱导CSF中IGF-1水平升高，但血清中IGF-1水平未升高。我们的结论是室管膜途径构成了一个有效的方法，在大脑中实施实验性IGF-1基因治疗。
Intracerebroventricular administration of the peptide insulin-like growth factor-1 (IGF-1) has been shown to be an effective neuroprotective strategy in the brain of different animal models, a major advantage being the achievement of high concentrations of IGF-1 in the brain without altering serum levels of the peptide. In order to exploit this therapeutic approach further, we used high performance recombinant adenoviral (RAd) vectors expressing their transgene under the control of the potent mouse cytomegalovirus immediate early (mCMV) promoter, to transduce brain ependymal cells with high efficiency and to achieve effective release of transgenic IGF-1 into the cerebrospinal fluid (CSF). We constructed RAd vectors expressing either the chimeric protein (TK/GFP)fus (green fluorescent protein fused to HSV1 thymidine kinase) or the cDNA encoding rat IGF-1, both driven by the mCMV promoter. The vectors were injected into the lateral ventricles of young rats and chimeric GFP expression in brain sections was assessed by fluorescence microscopy. The ependymal cell marker vimentin was detected by immunofluorescence and nuclei were labeled with the DNA dye DAPI. Blood and CSF samples were drawn at different times post vector injection. In all cerebral ventricles, vimentin immunoreactive cells of the ependyma were predominantly transduced by RAd-(TK/GFP)fus, showing nuclear and cytoplasmic expression of the transgene. For tanycytes (TK/GFP)fus expression was evident in their cytoplasmic processes as they penetrated deep into the hypothalamic parenchyma. Intracerebroventricular injection of RAd-IGF-1 induced high levels of IGF-1 in the CSF but not in serum. We conclude that the ependymal route constitutes an effective approach for implementing experimental IGF-1 gene therapy in the brain.
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影响因子：
5.1
作者：
Herenu, C. B.;Cristina, C.;Goya, R. G.
通讯作者：
Goya, R. G.
DOI：
10.1073/pnas.94.4.1287
发表时间：
1997-02-18
影响因子：
11.1
作者：
Ohashi, T;Watabe, K;Eto, Y
通讯作者：
Eto, Y
影响因子：
3.3
作者：
Markowska, AL;Mooney, M;Sonntag, WE
通讯作者：
Sonntag, WE
影响因子：
3.3
作者：
Guan, J;Miller, OT;Gluckman, PD
通讯作者：
Gluckman, PD
影响因子：
4.2
作者：
Paquin, A;Jaalouk, DE;Galipeau, J
通讯作者：
Galipeau, J