Noninvasive Monitoring of mRFP1-and mCherry-Labeled Oncolytic Adenoviruses in an Orthotopic Breast Cancer Model by Spectral Imaging
Noninvasive Monitoring of mRFP1-and mCherry-Labeled Oncolytic Adenoviruses in an Orthotopic Breast Cancer Model by Spectral Imaging
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DOI：
10.2310/7290.2010.00003
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发表时间：
2010-03-01
影响因子：
2.8
通讯作者：
Zinn, Kurt R.
中科院分区：
文献类型：
--
作者：
Borovjagin, Anton V.;McNally, Lacey R.;Zinn, Kurt R.
用荧光标记物对条件复制型腺病毒（CRAd）的基因衣壳进行标记提供了一种潜在的更准确的体内病毒治疗剂监测方法。用单体红色荧光蛋白1（mRFP 1）或其高级衍生物“mCherry”对感染性增强的CRAd（Ad 5/3，delta 24）的衣壳进行遗传标记，以评估每个红色荧光报告基因的实用性和CRAd的益处通过新的光谱成像方法在动物肿瘤模型中进行无创病毒追踪。通过与病毒衣壳蛋白IX的遗传融合将任一报道基因掺入CRAd颗粒中。瘤内注射后，通过光谱成像分析原位乳腺癌异种移植物中每种病毒的定位和复制，并通过定量聚合酶链反应进行验证。到第4天，肿瘤中的荧光增加高达2，000倍，并持续5至7周，显示出反映CRAd复制周期的振荡动力学。因此，衣壳标记结合光谱成像使得能够在报告基因转基因表达之前直接可视化和定量肿瘤中的CRAd颗粒。这允许非侵入性控制CRAd在肿瘤中的递送和分布，并促进病毒复制的定量评估。尽管mCherry作为成像标记物似乎上级mRFP 1，但两种报告者均显示出对CRAD成像应用的实用性。
Genetic capsid labeling of conditionally replicative adenoviruses (CRAds) with fluorescent tags offers a potentially more accurate monitoring of those virotherapy agents in vivo. The capsid of an infectivity-enhanced CRAd, Ad5/3, delta 24, was genetically labeled with monomeric red fluorescent protein 1 (mRFP1) or its advanced derivative, "mCherry," to evaluate the utility of each red fluorescent reporter and the benefit of CRAd capsid labeling for noninvasive virus tracking in animal tumor models by a new spectral imaging approach. Either reporter was incorporated into the CRAd particles by genetic fusion to the viral capsid protein IX. Following intratumoral injection, localization and replication of each virus in orthotopic breast cancer xenografts were analyzed by spectral imaging and verified by quantitative polymerase chain reaction. Fluorescence in tumors increased up to 2,000-fold by day 4 and persisted for 5 to 7 weeks, showing oscillatory dynamics reflective of CRAd replication cycles. Capsid labeling in conjunction with spectral imaging thus enables direct visualization and quantification of CRAd particles in tumors prior to the reporter transgene expression. This allows for noninvasive control of CRAd delivery and distribution in tumors and facilitates quantitative assessment of viral replication. Although mCherry appeared to be superior to mRFP1 as an imaging tag, both reporters showed utility for CRAd imaging applications.