Focal Delivery of AAV2/1-transgenes Into the Rat Brain by Localized Ultrasound-induced BBB Opening.
Focal Delivery of AAV2/1-transgenes Into the Rat Brain by Localized Ultrasound-induced BBB Opening.
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DOI：
10.1038/mtna.2012.64
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发表时间：
2013-02-19
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--
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药物和大分子向中枢神经系统（CNS）的递送受到血脑屏障（BBB）的阻碍。已经使用了几种方法来克服这种障碍，以促进各种CNS疾病的治疗。我们现在提出的结果表明，嵌合腺相关病毒2/1（AAV 2/1）颗粒含有LacZ基因的编码区有效地交付到大鼠脑后，通过聚焦超声血脑屏障开放后，在血管声学共振器的存在下，静脉（IV）管理。我们表明，转基因是正确和有效地表达在细胞位于附近的超声波焦点，特别是在附近的小血管和毛细血管。组织化学LacZ染色允许鉴定大量表达酶活性蛋白的细胞。使用双重免疫荧光（IF）与微管蛋白III和细菌LacZ的抗体，我们确定这些细胞主要是神经元。一小部分转导细胞被认为是神经胶质细胞，在IF中与星形胶质细胞标记物抗体反应阳性。这些结果表明，我们的方法允许一个非常具体的，本地化的，和有效的表达静脉注射转基因在大鼠脑超声诱导的血脑屏障开放。
Delivery of drugs and macromolecules to the central nervous system (CNS) is hindered by the blood–brain barrier (BBB). Several approaches have been used to overcome this hindrance to facilitate the treatment of various CNS diseases. We now present results showing that chimeric adeno-associated virus 2/1 (AAV2/1) particles containing the coding region for the LacZ gene are efficiently delivered into the rat brain upon intravenous (IV) administration after BBB opening by focused ultrasound in the presence of vascular acoustic resonators. We show that the transgene is correctly and efficiently expressed in cells located in the neighborhood of the insonated focus, especially in the vicinity of small vessels and capillaries. Histochemical LacZ staining allows the identification of large amounts of cells expressing the enzymatically active protein. Using double immunofluorescence (IF) with antibodies against tubulinIII and bacterial LacZ, we identified these cells to be mostly neurons. A small proportion of the transduced cells was recognized as glial cells, reacting positive in the IF with antibodies against astrocytic markers. These results demonstrate that our approach allows a very specific, localized, and efficient expression of intravenously administered transgenes in the brain of rats upon ultrasound-induced BBB opening.
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