Bimodal viral vectors and in vivo imaging reveal the fate of human neural stem cells in experimental glioma model
Bimodal viral vectors and in vivo imaging reveal the fate of human neural stem cells in experimental glioma model
复制标题
DOI：
10.1523/jneurosci.0296-08.2008
复制
发表时间：
2008-04-01
影响因子：
5.3
通讯作者：
Weissleder, Ralph
中科院分区：
文献类型：
--
作者：
Shah, Khalid;Hingtgen, Shawn;Weissleder, Ralph
将基因工程细胞移植到中枢神经系统为治疗多种神经系统疾病提供了巨大的潜力。监测转基因的表达水平并跟踪细胞功能和分布随时间的变化对于评估此类细胞的体内治疗功效至关重要。我们设计了含有荧光和生物发光标记蛋白不同组合之间融合的慢病毒载体，并使用生物发光成像和活体扫描显微镜实时研究体内患有神经胶质瘤的大脑中以细胞分辨率研究人类神经干细胞（hNSC）的命运。使用表达海肾荧光素酶 (Rluc)-DsRed2 或 GFP-Rluc 的恶性人神经胶质瘤模型，转导的 hNSC 被证明能够广泛向神经胶质瘤迁移，移植后 10 天，hNSC 就开始在神经胶质瘤中生长。此外，转导的 hNSC 在患有神经胶质瘤的小鼠中比在正常大脑中存活更长时间，但在体内并不能调节神经胶质瘤的进展。这些研究证明了双峰病毒载体和实时成像在评估疾病模型中 NSC 的命运方面的实用性，从而为加速 CNS 疾病的细胞疗法提供了一个平台。
Transplantation of genetically engineered cells into the CNS offers immense potential for the treatment of several neurological disorders. Monitoring expression levels of transgenes and following changes in cell function and distribution over time is critical in assessing therapeutic efficacy of such cells in vivo. We have engineered lentiviral vectors bearing fusions between different combinations of fluorescent and bioluminescent marker proteins and used bioluminescence imaging and intravital-scanning microscopy in real time to study the fate of human neural stem cells ( hNSCs) at a cellular resolution in glioma-bearing brains in vivo. Using Renilla luciferase (Rluc)-DsRed2 or GFP-Rluc-expressing malignant human glioma model, transduced hNSCs were shown to migrate extensively toward gliomas, with hNSCs populating gliomas at 10 d after transplantation. Furthermore, transduced hNSCs survived longer in mice with gliomas than in normal brain, but did not modulate glioma progression in vivo. These studies demonstrate the utility of bimodal viral vectors and real-time imaging in evaluating fate of NSCs in diseased models and thus provide a platform for accelerating cell-based therapies for CNS disorders.