Biodistribution and toxicity studies of VSVG-pseudotyped lentiviral vector after intravenous administration in mice with the observation of in vivo transduction of bone marrow
Biodistribution and toxicity studies of VSVG-pseudotyped lentiviral vector after intravenous administration in mice with the observation of in vivo transduction of bone marrow
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DOI：
10.1006/mthe.2002.0630
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发表时间：
2002-07-01
影响因子：
12.4
通讯作者：
Whitley, CB
中科院分区：
文献类型：
--
作者：
Pan, D;Gunther, R;Whitley, CB
慢病毒载体可以在多种细胞类型中赋予高水平的基因转移和转基因表达。然而，静脉给药后的生物分布和毒性尚未报道。为了解决这些生物分布和毒性的问题，通过尾静脉注射将基于HIV-1的载体HR'cmvGFP施用给正常BALB/c小鼠。通过实时定量PCR（QPCR）试验评价了9个不同器官和骨髓，该试验能够在宽范围内定量（5-log倍），以检测每10（5）个细胞中少至1个绿色荧光蛋白基因（GFP）拷贝。载体给药后4天，在所有动物的肝脏、脾脏和骨髓中观察到高水平的转基因和基因表达。注射后40天，肝脏和脾脏中的GFP水平下降，但骨髓中的转基因水平一直很高。这一发现与通过荧光激活细胞分选（FACS）分析在外周血中观察到的GFP频率和表达水平的增加一致。在所有其他器官中检测到0至1%的转基因。在检查的任何组织中均未发现可归因于载体的显著病理学病变。静脉内载体给药后骨髓转导的观察表明干细胞基因治疗的体内方法的可能性。
Lentiviral vectors can confer high levels of gene transfer and transgene expression in a variety of cell types. However, the biodistribution and toxicity after intravenous administration have not been reported. To address these issues of biodistribution and toxicity, an HIV-1-based vector, HR'cmvGFP, was administered to normal BALB/c mice by tail-vein injection. Nine different organs and bone marrow were evaluated by real-time quantitative PCR (QPCR) assay capable of a broad range of quantitation (5-log fold) to detect as few as one copy of the green fluorescent protein gene (GFP) per 10(5) cells. Four days after vector administration, high levels of transgene and gene expression were observed in liver, spleen, and bone marrow in all animals. By 40 days after injection, GFP levels had decreased in liver and spleen, but bone marrow exhibited a consistently high level of transgene. This finding was consistent with the increase in both GFP frequency and expression levels observed in peripheral blood by fluorescence-activated cell-sorting (FACS) analysis. Between 0 and 1% transgene was detected in all other organs. No significant pathologic lesions were found attributable to vector in any of the tissues examined. The observation of bone marrow transduction after intravenous vector administration suggests the possibility of an in vivo approach to stem cell gene therapy.