A novel hybrid baculovirus-adeno-associated viral vector-mediated radionuclide reporter gene imaging system for stem cells transplantation monitoring
A novel hybrid baculovirus-adeno-associated viral vector-mediated radionuclide reporter gene imaging system for stem cells transplantation monitoring
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一种新型混合杆状病毒-腺相关病毒载体介导的放射性核素报告基因成像系统，用于干细胞移植监测
DOI：
10.1007/s00253-014-6162-0
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发表时间：
2015-02-01
影响因子：
5
通讯作者：
Zhang，Yifan
中科院分区：
文献类型：
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作者：
Pan，Yu;Yin，Hongyan;Zhang，Yifan
含有增强型绿色荧光蛋白（eGFP）报告基因或人钠碘同向转运体（hNIS）报告基因的杂合杆状病毒-腺相关病毒（BV-AAV），所述报告基因的侧翼为来自AAV的反向末端重复序列（ITR）构建了BV-CMV-eGFP-ITR和BV-CMV-hNIS-ITR两种转基因载体，并探讨了利用杂合BV-AAV转基因载体介导hNIS报告基因显像用于骨髓造血干细胞监测的可行性。骨髓间充质干细胞（BM-MSCs）移植治疗作为放射性核素报告基因显像的新生物技术平台。结果表明，BV-CMV-eGFP-ITR对BM-MSCs的感染率达到84.25 ± 1.38%，对BM-MSCs无明显不良影响。125 I-和99 mTcO 4-摄取试验表明BA-AAV介导的hNIS在感染的BM-MSCs中诱导的放射性核素蓄积是高效的。此外，受感染的BM-MSCs细胞数量与125 I-蓄积量之间存在强相关性（R2= 0.9026）。micro-SPECT/CT显像显示BV-CMV-hNIS-ITR感染的BM-MSCs体内有效地蓄积放射性碘，在移植部位表现出明显的放射性示踪剂蓄积。进一步定量分析显示30 min可能是最佳成像时间点。此外，所揭示的高靶/个体器官背景比也支持BV-AAV介导的hNIS报告基因成像用于监测大多数常用移植部位中的BM-MSCs移植的可行性，从而突出了放射性核素报告基因成像用于干细胞移植治疗的前景生物技术平台。
Hybrid baculovirus-adeno-associated virus (BV-AAV) containing enhanced green fluorescent protein (eGFP) reporter gene or human sodium-iodide symporter (hNIS) reporter gene flanked by inverted terminal repeats (ITRs) derived from AAV (BV-CMV-eGFP-ITR and BV-CMV-hNIS-ITR) were constructed and used to investigate the feasibility of using hybrid BV-AAV transgenic vector to mediate hNIS reporter gene imaging for monitoring bone marrow-derived mesenchymal stem cells (BM-MSCs) transplantation therapy as a novel biotechnological platform in radionuclide reporter gene imaging. The results showed that the infection efficiency of BV-CMV-eGFP-ITR in BM-MSCs reached 84.25 ± 1.38 %, and there were no obvious adverse effects on BM-MSCs. The125I−and99mTcO4−uptake assays showed that the radionuclide accumulation induced by BA-AAV-mediated hNIS was highly efficient in infected BM-MSCs. Furthermore, there was a robust correlation between the infected BM-MSCs cell number and the125I−accumulation amount (R2= 0.9026). The micro-SPECT/CT imaging showed that BV-CMV-hNIS-ITR-infected BM-MSCs accumulated radioiodine efficiently in vivo, exhibiting obvious radiotracer accumulation in transplantation sites. Further quantitative analysis revealed that 30 min might be the optimal imaging time point. Moreover, the revealed high target/individual organ background ratios also supported the feasibility of BV-AAV-mediated hNIS reporter gene imaging for monitoring BM-MSCs transplantation in most of commonly used transplantation sites, thus highlighting this promise biotechnological platform in radionuclide reporter gene imaging for stem cell transplantation therapy.