Franchi Federico, Peterson Karen M, Paulmurugan Ramasamy, Folmes Clifford, Lanza Ian R, Lerman Amir, Rodriguez-Porcel Martin
Division of Cardiovascular Diseases, Department of Internal Medicine, Mayo Clinic, 200 First St. SW, Rochester, MN, 55905, USA.
Department of Radiology and Molecular Imaging Program at Stanford (MIPS), Stanford University School of Medicine, Stanford, CA, USA.
Mol Imaging Biol. 2016 Aug;18(4):510-8. doi: 10.1007/s11307-016-0929-x.
Mitochondria are a gatekeeper of cell survival and mitochondrial function can be used to monitor cell stress. Here we validate a pathway-specific reporter gene to noninvasively image the mitochondrial function of stem cells.
We constructed a mitochondrial sensor with the firefly luciferase (Fluc) reporter gene driven by the NQO1 enzyme promoter. The sensor was introduced in stem cells and validated in vitro and in vivo, in a mouse model of myocardial ischemia/reperfusion (IR).
The sensor activity showed an inverse relationship with mitochondrial function (R (2) = -0.975, p = 0.025) and showed specificity and sensitivity for mitochondrial dysfunction. In vivo, NQO1-Fluc activity was significantly higher in IR animals vs. controls, indicative of mitochondrial dysfunction, and was corroborated by ex vivo luminometry.
Reporter gene imaging allows assessment of the biology of transplanted mesenchymal stromal cells (MSCs), providing important information that can be used to improve the phenotype and survival of transplanted stem cells.
线粒体是细胞存活的守门人，线粒体功能可用于监测细胞应激。在此，我们验证一种途径特异性报告基因，以无创成像干细胞的线粒体功能。
我们构建了一种线粒体传感器，其萤火虫荧光素酶（Fluc）报告基因由NQO1酶启动子驱动。该传感器被导入干细胞，并在体外以及心肌缺血/再灌注（IR）小鼠模型中进行体内验证。
传感器活性与线粒体功能呈负相关（R (2) = -0.975，p = 0.025），并对线粒体功能障碍具有特异性和敏感性。在体内，IR动物的NQO1-Fluc活性显著高于对照组，表明线粒体功能障碍，体外发光测定法证实了这一点。
报告基因成像可评估移植的间充质基质细胞（MSC）的生物学特性，提供可用于改善移植干细胞表型和存活率的重要信息。
