College of Chemistry, Chemical Engineering and Materials Science, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong , Shandong Normal University , Jinan 250014 , P. R. China.
Medicine & Pharmacy Research Center , Binzhou Medical University , Shandong , Yantai 264003 , P. R. China.
ACS Sens. 2019 Mar 22;4(3):733-739. doi: 10.1021/acssensors.9b00026. Epub 2019 Feb 28.
A growing body of evidence indicates that micropeptides encoded by long noncoding RNAs (lncRNAs) act independently or as regulators of larger proteins in fundamental biological processes, especially in the maintenance of cellular homeostasis. However, due to their small size and low intracellular expression, visual monitoring of micropeptides in living cells is still a challenge. In this work, we have designed and synthesized an aptamer-based near-infrared fluorescence nanoprobe for fluorescence imaging of phospholamban (PLN), which is an intracellular micropeptide that affects calcium homeostasis, and is closely associated with human heart failure in the clinic. The nanoprobe could respond specifically to PLN with excellent selectivity, high sensitivity, good nuclease stability, and biocompatibility, and it was successfully applied for imaging of changes in PLN levels in cardiomyocytes and in frozen sections of heart tissues. Further combined with clinical myocardial biopsy, we believe that the developed nanoprobe should be of great significance in later molecular pathology study of heart failure, which may help with diagnosis of early heart failure in the future. More importantly, for the first time nanoprobes were applied to visually monitor the changes of micropeptides in living cells and in frozen tissue sections, and the design concept of the aptamer-based nanoprobe can be extended to fluorescence detection of other micropeptides.
越来越多的证据表明,长非编码 RNA(lncRNA)编码的微肽独立或作为更大蛋白质的调节剂在基本的生物过程中发挥作用,尤其是在维持细胞内环境稳定方面。然而,由于其体积小、细胞内表达水平低,因此在活细胞中对微肽进行可视化监测仍然是一个挑战。在这项工作中,我们设计并合成了一种基于适体的近红外荧光纳米探针,用于对影响钙稳态的细胞内微肽——肌球蛋白结合蛋白 C(PLN)进行荧光成像,PLN 与临床上的人类心力衰竭密切相关。该纳米探针对 PLN 具有出色的选择性、高灵敏度、良好的核酸酶稳定性和生物相容性,可以成功地用于成像心肌细胞中 PLN 水平的变化以及心脏组织的冷冻切片。进一步结合临床心肌活检,我们相信,开发的纳米探针在心力衰竭的后期分子病理学研究中具有重要意义,这可能有助于未来早期心力衰竭的诊断。更重要的是,该纳米探针首次被应用于可视化监测活细胞和冷冻组织切片中微肽的变化,基于适体的纳米探针的设计理念可以扩展到其他微肽的荧光检测。