Wang Jian, Hong Donghui, Li Jili, Wang Linlin, Xie Yuqi, Da Jun, Liu Yanlan
Molecular Science and Biomedicine Laboratory (MBL), State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Aptamer Engineering Center of Hunan Province, Hunan University, Changsha, Hunan 410082, China.
ACS Meas Sci Au. 2024 Aug 31;4(5):577-584. doi: 10.1021/acsmeasuresciau.4c00045. eCollection 2024 Oct 16.
Simultaneous detection of multiple biomarkers is crucial to achieve specific and dynamic analysis of cellular senescence, given its intrinsic high heterogeneity. Current approaches for senescence detection largely rely on fluorescence imaging, but fluorescent probes inevitably suffer from issues including autofluorescence and spectral overlap when being applied for the simultaneous detection of multiple biomarkers. Herein, we report an alternative strategy and design activatable multiplexed senoprobes based on F NMR for dynamic monitoring of cellular senescence. Differing from previous approaches, our strategy has two unique advantages. First, this strategy utilizes the changes in the F chemical shift as the signal output, which features by its fingerprint and quantifiable characters, thereby significantly enhancing the detection throughput toward biomarkers with minimized spectral overlapping. Second, the background signal is minimized, benefiting from the extremely low abundance of F in biological samples, and the detection accuracy can thus be improved. As a proof of concept, two activatable F NMR molecular probes are synthesized that specially respond to two key senescence-associated biomarkers (β-gal and ROS) and have been successfully demonstrated for dynamical and quantitative assessment of the changes of these biomarkers in different cellular models of senescence, without causing obvious cytotoxicity. Owing to the flexible molecular design, this work may offer a useful platform to create diversified F NMR senoprobes for deep understanding of cellular senescence across a wide range of aging-related diseases.
鉴于细胞衰老具有内在的高度异质性，同时检测多种生物标志物对于实现细胞衰老的特异性和动态分析至关重要。当前用于衰老检测的方法主要依赖于荧光成像，但在应用于同时检测多种生物标志物时，荧光探针不可避免地存在诸如自发荧光和光谱重叠等问题。在此，我们报告了一种替代策略，并基于¹⁹F NMR设计了可激活的多重衰老探针，用于动态监测细胞衰老。与先前的方法不同，我们的策略具有两个独特的优势。首先，该策略利用¹⁹F化学位移的变化作为信号输出，其具有指纹特征和可量化特性，从而以最小的光谱重叠显著提高了对生物标志物的检测通量。其次，由于生物样品中¹⁹F的丰度极低，背景信号被最小化，因此可以提高检测准确性。作为概念验证，合成了两种可激活的¹⁹F NMR分子探针，它们专门响应两种关键的衰老相关生物标志物（β-半乳糖苷酶和活性氧），并已成功用于动态和定量评估这些生物标志物在不同衰老细胞模型中的变化，且不会引起明显的细胞毒性。由于灵活的分子设计，这项工作可能提供一个有用的平台，以创建多样化的¹⁹F NMR衰老探针，用于深入了解广泛的衰老相关疾病中的细胞衰老。
