Tailored transgene expression to specific cell types in the central nervous system after peripheral injection with AAV9
Tailored transgene expression to specific cell types in the central nervous system after peripheral injection with AAV9
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DOI：
10.1038/mtm.2016.81
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发表时间：
2016-01-01
影响因子：
4.7
通讯作者：
Hudry, Eloise
中科院分区：
文献类型：
--
作者：
Dashkoff, Jonathan;Lerner, Eli P.;Hudry, Eloise
某些腺相关病毒(AAV)载体在静脉注射后跨越血脑屏障的能力为非侵入性脑传递提供了独特的机会。然而，如果没有一个量身定制的系统，使用外周途径注射可能会导致非靶细胞或器官中不希望看到的转基因表达。为了改进这一方法，本研究描述了静脉注射成年小鼠(2x10(13)vg/kg)后，在星形胶质细胞(胶质纤维酸性(GFA)蛋白)或神经元(突触素(Syn))启动子下表达绿色荧光蛋白(GFP)的新的自我互补AAV9(ScAAV9)的转导特征。ScAAV9-GFA-GFP和scAAV9-Syn-GFP分别健壮地转导星形胶质细胞(11%)和神经元(17%)，无异常表达泄漏。有趣的是，尽管带有scAAV9-GFA-GFP的星形胶质细胞表达GFP的百分比与使用scAAV9-CBA-GFP(广谱活性启动子)的星形胶质细胞相似，但使用scAAV9-Syn-GFP的神经元表达GFP的百分比显著高于scAAV9-Syn-GFP。在脊髓内可见GFP阳性的兴奋性神经元和抑制性神经元，以及运动神经元。此外，注射scAAV9-GFA-GFP后，激活的(GFAP阳性的)和静息的(GFAP阴性的)星形胶质细胞都表达报告基因。这些数据充分表征了静脉注射后携带神经元和星形胶质细胞选择性启动子的AAVs的基因表达特异性，这将被证明有助于中枢神经系统(CNS)的基因治疗方法，其中外周表达的转基因是一个关注的问题。
The capacity of certain adeno-associated virus (AAV) vectors to cross the blood-brain barrier after intravenous delivery offers a unique opportunity for noninvasive brain delivery. However, without a well-tailored system, the use of a peripheral route injection may lead to undesirable transgene expression in nontarget cells or organs. To refine this approach, the present study characterizes the transduction profiles of new self-complementary AAV9 (scAAV9) expressing the green fluorescent protein (GFP) either under an astrocyte (glial fibrillary acidic (GFA) protein) or neuronal (Synapsin (Syn)) promoter, after intravenous injection of adult mice (2 x10(13) vg/kg). ScAAV9-GFA-GFP and scAAV9-Syn-GFP robustly transduce astrocytes (11%) and neurons (17%), respectively, without aberrant expression leakage. Interestingly, while the percentages of GFP-positive astrocytes with scAAV9-GFA-GFP are similar to the performances observed with scAAV9-CBA-GFP (broadly active promoter), significant higher percentages of neurons express GFP with scAAV9-Syn-GFP. GFP-positive excitatory as well as inhibitory neurons are observed, as well as motor neurons in the spinal cord. Additionally, both activated (GFAP-positive) and resting astrocytes (GFAP-negative) express the reporter gene after scAAV9-GFA-GFP injection. These data thoroughly characterize the gene expression specificity of AAVs fitted with neuronal and astrocyte-selective promoters after intravenous delivery, which will prove useful for central nervous system (CNS) gene therapy approaches in which peripheral expression of transgene is a concern.