Engineered HSV vector achieves safe long-term transgene expression in the central nervous system.
Engineered HSV vector achieves safe long-term transgene expression in the central nervous system.
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DOI：
10.1038/s41598-017-01635-1
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发表时间：
2017-05-04
影响因子：
4.6
通讯作者：
Glorioso JC
中科院分区：
文献类型：
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作者：
Verlengia G;Miyagawa Y;Ingusci S;Cohen JB;Simonato M;Glorioso JC
此前，我们报道了一系列新的高度缺陷的1型单纯疱疹病毒(HSV-1)载体，这些载体在功能上缺乏所有病毒即刻早期(IE)基因，导致实际上缺乏病毒基因的表达。然而，插入到载体中的报告基因盒被来自病毒潜伏期基因位点的边界元件包围，在非神经元细胞中显示出高的、持久的报告基因活性，而插入到缺失的ICP4基因位点的独立表达盒几乎保持沉默。与非神经细胞相比，我们在这里展示了ICP4基因盒允许报告基因在不同的大鼠脑区域立体定向注射后在不同的神经元中表达；转基因在海马区的表达持续了6个月，基本上局限于神经元。没有观察到神经细胞毒性或诱导炎性细胞浸润的证据。位于基因间隔区的独立报告基因盒保持沉默，表明转基因启动子和/或插入位点对持续表达至关重要。这些发现表明，该载体适合于对需要表达大的和/或多个治疗性转基因的中枢神经系统疾病进行治疗干预。
Previously we reported a new series of highly defective herpes simplex virus type 1 (HSV-1) vectors that were functionally devoid of all viral immediately early (IE) genes, resulting in virtual absence of viral gene expression. Nevertheless, a reporter gene cassette inserted into the vector flanked by boundary elements from the viral latency locus showed high, persistent reporter gene activity in non-neuronal cells while an independent expression cassette inserted into a deleted ICP4 locus remained almost silent. In contrast to non-neuronal cells, we show here that the ICP4 locus cassette permitted robust reporter gene expression in a diversity of neurons following stereotactic injection of different rat brain regions; transgene expression in the hippocampus lasted up to 6 months and was essentially restricted to neurons. No evidence of neuronal cell toxicity or induction of inflammatory cell infiltrates was observed. An independent reporter gene cassette located in an intergenic region remained silent, indicating that the transgene promoter and/or insertion site are critical for sustained expression. These findings suggest the suitability of this vector for therapeutic intervention into diseases of the central nervous system that require the expression of large and/or multiple therapeutic transgenes.