细胞应激过程线粒体响应的成像分析

Subcellular organelles are the fundamental entities for cell functionality. It is of biomedical interest to define organelle responses under pathological conditions. Mitochondria are critical for cell metabolism, autophagy, apoptosis, and oncogenesis, etc. To date, the roles of mitochondrial parameters (trans-membrane electrical potential, subcellular localization, and organelle-interaction, ROS generation) have been largely unexplored in these processes..We will develop mitochondria-targeted ratiometric fluorescence probes for tracking changes of mitochondrial parameters in the cell signaling events related to cell death, autophagy, and inflammation. Specifically, hetero-organelle-responsive dual colored probes will be used to define the loss of trans-membrane potential of mitochondria, ratiometric probes for covalent labeling of mitochondria to explore the engulfment and digestion of damaged mitochondria by lysosomes and spatiotemporal interaction of organelles with phagosome, golgi, and lysosomes; ROS-responsive probe to determine ROS generated in mitochondria in stressed cells.. Finally, we will use ratiometric fluorescence imaging to explore the effects of antibiotics, and mitochondria-targeted anti-cancer drugs on mitochondrial parameters. This work will provide novel analytical techniques for investigation of mitochondria biology and offer a new perspective to explore the efficacy of drugs for mitochondria-associated diseases.

线粒体是一种具有跨膜电势的、提供能量的细胞器。线粒体的膜电势与细胞能量代谢、氧化稳态、自噬、多种神经退化疾病密切关联。课题发展线粒体靶向、兼容不同生物过程的比率荧光探针；用于分析细胞应激过程线粒体变化（膜电势、位置、降解、活性氧产生），阐明线粒体变化与应激反应关联（死亡、炎症等）；分析影响线粒体功能的药物机理。这将拓展小分子探针成像技术在细胞器研究中应用，并为线粒体相关疾病治疗提供新科学依据。

线粒体参与多种重要生理过程。受损细胞器对疾病发生发展有重要影响，如线粒体关联神经退化疾病。发展能够保护线粒体功或破坏癌细胞线粒体功能的新方法已经成为临床医学研究重要领域。我们发展细胞器导向的有机反应，在线粒体内组装比率荧光探针，实现应激或受损线粒体的精准稳定跟着，分析应激过程线粒体变化规律，发展： 1）绿-红荧光变换方法灵敏分析mitopahgy， 筛选评估线粒体靶向药物及其影响线粒体功能的药效；2）基于糖分选通路的应激溶酶体分析新方法，分析应激线粒体与溶酶体互动；3）细胞器内生物正交反应：新型线粒体探针用于应激线粒体的膜通透性分析；4）细胞器导向线粒体内膜代谢标记：铁死亡过程线粒体自噬的发现，5）溶酶体靶向的唾液酸探针同于衰老、炎症条件下细胞器的研究；6）膜电势驱动的线粒体DNA荧光标记新方法，7）双细胞器靶向的双ROS产生的光动力学疗法，用于肿瘤治疗与荧光辅助肿瘤切除。本项目发展的分子探针初步实现对重大疾病关联线粒体变化进行可视化研究，将为线粒体相关生物医学研究提供新方法。