Intra-cisterna magna delivery of an AAV vector with the GLUT1 promoter in a pig recapitulates the physiological expression of SLC2A1
Intra-cisterna magna delivery of an AAV vector with the GLUT1 promoter in a pig recapitulates the physiological expression of SLC2A1
复制标题
在猪的小脑池内递送带有 GLUT1 启动子的 AAV 载体，重现了 SLC2A1 的生理表达
DOI：
10.1038/s41434-020-00203-z
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发表时间：
2021
期刊：
影响因子：
5.1
通讯作者：
Yamagata Takanori
中科院分区：
文献类型：
--
作者：
Nakamura Sachie;Osaka Hitoshi;Muramatsu Shin-ichi;Takino Naomi;Ito Mika;Jimbo Eriko F.;Watanabe Chika;Hishikawa Shuji;Nakajima Takeshi;Yamagata Takanori
葡萄糖转运蛋白1缺乏综合征（GLUT1DS）是由编码GLUT1的slc2a1单倍体功能不足引起的，导致己糖转运到大脑受损。此前，我们构建了酪氨酸突变体AAV9/3载体，其中slc2a1在内源性GLUT1启动子（AAV-GLUT1）的控制下表达，并证实脑室注射AAV-GLUT1后GLUT1缺陷小鼠的运动功能和脑脊液葡萄糖水平得到改善。为了准备临床应用，我们检测了大池内注射AAV-GFP（酪氨酸突变型带有CMV启动子的AAV9/3-GFP）和AAV-GLUT1后转基因的表达。我们将AAV-GFP或AAV-GLUT1 （1.63 × 1012个载体基因组/kg）注射到猪大池中，比较不同启动子的活性。注射AAV-GFP后，外源性GFP在脑和周围器官的广泛区域表达。注射AAV-GLUT1后，外源性GLUT1主要在脑内表达。在细胞水平上，外源性GLUT1主要在内皮细胞中表达，其次是胶质细胞和神经元，这与CMV启动子以神经元为主表达GFP形成对比。我们认为大池内注射AAV-GLUT1是一种可行的GLUT1DS基因治疗方法。
Glucose transporter 1 deficiency syndrome (GLUT1DS) is caused by haplo-insufficiency ofSLC2A1, which encodes GLUT1, resulting in impaired hexose transport into the brain. Previously, we generated a tyrosine-mutant AAV9/3 vector in whichSLC2A1was expressed under the control of the endogenous GLUT1 promoter (AAV-GLUT1), and confirmed the improved motor function and cerebrospinal fluid glucose levels ofGlut1-deficient mice after cerebroventricular injection of AAV-GLUT1. In preparation for clinical application, we examined the expression of transgenes after intra-cisterna magna injection of AAV-GFP (tyrosine-mutant AAV9/3-GFP with the CMV promoter) and AAV-GLUT1. We injected AAV-GFP or AAV-GLUT1 (1.63 × 1012vector genomes/kg) into the cisterna magna of pigs to compare differential promoter activity. After AAV-GFP injection, exogenous GFP was expressed in broad areas of the brain and peripheral organs. After AAV-GLUT1 injection, exogenous GLUT1 was expressed predominantly in the brain. At the cellular level, exogenous GLUT1 was mainly expressed in the endothelium, followed by glia and neurons, which was contrasted with the neuronal-predominant expression of GFP by the CMV promotor. We consider intra-cisterna magna injection of AAV-GLUT1 to be a feasible approach for gene therapy of GLUT1DS.