Li Nuan, Wang Peng, Xie Yuanyuan, Wang Bin, Zhu Chenzhuo, Xue Le, Han Xiaofeng, Gu Ning, Sun Jianfei
Jiangsu Key Laboratory of Biomaterials and Devices, School of Biological Sciences and Medical Engineering, Southeast University, Nanjing 210096, China.
Jiangsu Key Laboratory of Biomaterials and Devices, School of Biological Sciences and Medical Engineering, Southeast University, Nanjing 210096, China; Department of Sports Medicine and Adult Reconstructive Surgery, the Affiliated Drum Tower Hospital of Nanjing University, Nanjing 210008, China.
Acta Biomater. 2023 Dec;172:309-320. doi: 10.1016/j.actbio.2023.09.039. Epub 2023 Sep 29.
Here, we propose for the first time the evaluation of magnetosensitive clMagR/clCry4 as a magnetic resonance imaging (MRI) reporter gene that imparts sensitivity to endogenous contrast in eukaryotic organisms. Using a lentiviral vector, we introduced clMagR/clCry4 into C57BL/6 mice-derived bone marrow mesenchymal stem cells (mBMSCs), which could specifically bind with iron, significantly affected MRI transverse relaxation, and generated readily detectable contrast without adverse effects in vivo. Specifically, clMagR/clCry4 makes mBMSCs beneficial for enhancing the sensitivity of MRI-R for iron-bearing granules, in which cells recruit exogenous iron and convert these stores into an MRI-detectable contrast; this is not achievable with control cells. Additionally, Prussian blue staining was performed together with ultrathin cell slices to provide direct evidence of natural iron-bearing granules being detectable on MRI. Hence, it was inferred that the sensitivity of MRI detection should be correlated with clMagR/clCry4 and exogenous iron. Taken together, the clMagR/clCry4 has great potential as an MRI reporter gene. STATEMENT OF SIGNIFICANCE: In this study, we propose the evaluation of magnetosensitive clMagR/clCry4 as an MRI reporter gene, imparting detection sensitivity to eukaryotic mBMSCs for endogenous contrast. At this point, the clMagR and clCry4 were located within the cytoplasm and possibly influence each other. The clMagR/clCry4 makes mBMSCs beneficial for enhancing the sensitivity of MRI-R for iron-bearing granules, in which protein could specifically bind with iron and convert these stores into MRI-detectable contrast; this is not achieved by control cells. The viewpoint was speculated that the clMagR/clCry4 and exogenous iron were complementary to each other. Additionally, Prussian blue staining was performed together with TEM observations to provide direct evidence that the iron-bearing granules were sensitive to MRI.
在此,我们首次提出将磁敏蛋白clMagR/clCry4评估为一种磁共振成像(MRI)报告基因,该基因可赋予真核生物对内源性对比剂的敏感性。我们使用慢病毒载体将clMagR/clCry4导入源自C57BL/6小鼠的骨髓间充质干细胞(mBMSC)中,其能够特异性结合铁,显著影响MRI横向弛豫,并在体内产生易于检测的对比剂且无不良影响。具体而言,clMagR/clCry4使mBMSC有利于提高MRI对含铁颗粒的检测灵敏度,其中细胞募集外源性铁并将这些储存物转化为MRI可检测的对比剂;而对照细胞无法做到这一点。此外,将普鲁士蓝染色与超薄细胞切片一起进行,以提供天然含铁颗粒在MRI上可检测的直接证据。因此,据推断MRI检测的灵敏度应与clMagR/clCry4和外源性铁相关。综上所述,clMagR/clCry4作为MRI报告基因具有巨大潜力。重要性声明:在本研究中,我们提出将磁敏蛋白clMagR/clCry4评估为一种MRI报告基因,赋予真核mBMSC对内源性对比剂的检测灵敏度。此时,clMagR和clCry4位于细胞质内且可能相互影响。clMagR/clCry4使mBMSC有利于提高MRI对含铁颗粒的检测灵敏度,其中该蛋白可特异性结合铁并将这些储存物转化为MRI可检测的对比剂;对照细胞无法实现这一点。据推测,clMagR/clCry4与外源性铁相互补充。此外,将普鲁士蓝染色与透射电镜观察一起进行,以提供含铁颗粒对MRI敏感的直接证据。