Wu Joseph C, Spin Joshua M, Cao Feng, Lin Shuan, Xie Xiaoyan, Gheysens Olivier, Chen Ian Y, Sheikh Ahmad Y, Robbins Robert C, Tsalenko Anya, Gambhir Sanjiv S, Quertermous Tom
Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, USA.
Physiol Genomics. 2006 Mar 13;25(1):29-38. doi: 10.1152/physiolgenomics.00254.2005. Epub 2006 Jan 3.
Stem cell therapy offers exciting promise for treatment of ischemic heart disease. Recent advances in molecular imaging techniques now allow investigators to monitor cell fate noninvasively and repetitively. Here we examine the effects of a triple-fusion reporter gene on embryonic stem (ES) cell transcriptional profiles. Murine ES cells were stably transfected with a self-inactivating lentiviral vector carrying a triple-fusion (TF) construct consisting of fluorescence, bioluminescence, and positron emission tomography (PET) reporter genes. Fluorescence-activated cell sorting (FACS) analysis allowed isolation of stably transfected populations. Microarray studies comparing gene expression in nontransduced control ES cells vs. stably transduced ES cells expressing triple fusion (ES-TF) revealed some increases in transcriptional variability. Annotation analysis showed that ES-TF cells downregulated cell cycling, cell death, and protein and nucleic acid metabolism genes while upregulating homeostatic and anti-apoptosis genes. Despite these transcriptional changes, expression of the TF reporter gene had no significant effects on ES cell viability, proliferation, and differentiation capability. Importantly, transplantation studies in murine myocardium demonstrated the feasibility of tracking ES-TF cells in living subjects using bioluminescence and PET imaging. Taken together, this is the first study to analyze in detail the effects of reporter genes on molecular imaging of ES cells.
干细胞疗法为缺血性心脏病的治疗带来了令人兴奋的前景。分子成像技术的最新进展使研究人员能够无创且重复地监测细胞命运。在此，我们研究了三融合报告基因对胚胎干细胞（ES细胞）转录谱的影响。用携带由荧光、生物发光和正电子发射断层扫描（PET）报告基因组成的三融合（TF）构建体的自失活慢病毒载体稳定转染小鼠ES细胞。荧光激活细胞分选（FACS）分析可分离出稳定转染的细胞群体。通过微阵列研究比较未转导的对照ES细胞与表达三融合的稳定转导ES细胞（ES-TF）中的基因表达，发现转录变异性有所增加。注释分析表明，ES-TF细胞下调了细胞周期、细胞死亡以及蛋白质和核酸代谢相关基因，同时上调了稳态和抗凋亡基因。尽管有这些转录变化，但TF报告基因的表达对ES细胞的活力、增殖和分化能力没有显著影响。重要的是，在小鼠心肌中的移植研究证明了使用生物发光和PET成像在活体动物中追踪ES-TF细胞的可行性。综上所述，这是第一项详细分析报告基因对ES细胞分子成像影响的研究。
