亚细胞结构水平上活性小分子协同调控细胞功能的荧光成像研究

In recent years, many important breakthroughs have been achieved in the cell communication. However, as the physical basis, studies on correlation of subcellular distribution of biologically active small molecules and their coordinated regulation of cell functions has not attracted enough attention, which severely hinders the intensive understanding of signaling systems and cellular actions. Various intracellular reactive molecules in single organelle, especially active small molecules (reactive oxygen species, ions, and so on), or various/same reactive small molecules in different organelles, are closely associated with the cellular physiological and pathological processes, and synergistically regulate cellular actions spatially and temporally. In this proposal, we plan to design and synthesize organic fluorescent probes or fluorescent nanoprobes with high sensitivity, precise organelle localization and specific recognition to active small molecules, and develop in situ, real-time, dynamic and high-resolution fluorescent imaging methods at the subcellular level, which could reveal the related variation roles of different reactive small molecules in single organelle and various/same reactive small molecules in different organelles in the process of incidence, development and therapy of major diseases, such as cancer and diabetes, explore the synergetic regulation role of active small molecules toward cellular functions. Finally, these studies will provide new theoretical foundations and new avenues for in-depth understanding the nature of life and molecular mechanisms, as well as diagnosis and treatment in the incidence and development of major diseases.

尽管细胞通讯的研究取得了众多突破，但是作为物质基础的不同生物活性分子在亚细胞结构水平浓度分布的相关性，及对细胞功能的协同调控作用尚未引起重视，严重阻碍了对细胞功能的深入了解。细胞内单一细胞器的不同活性分子尤其是活性小分子（活性氧自由基、阴阳离子等），或不同细胞器的不同/相同活性小分子随着细胞生理、病理过程而发生紧密关联，在时空上整体协同调控细胞行为。本项目拟设计合成或组装高灵敏度、细胞器精确定位、特异性识别活性小分子的有机分子或纳米荧光探针，在亚细胞结构水平上，发展具有多水平、多层次的原位、实时、动态的高分辨荧光成像方法，揭示在肿瘤、糖尿病等重大疾病的发生发展及治疗过程中，单一细胞器的不同活性小分子、不同细胞器的不同/相同活性小分子在时间、空间的关联变化规律，探索活性小分子对细胞功能的协同调控作用，最终为深入理解生命现象本质、重大疾病发生发展的分子机制及其诊治提供新的理论依据及新途径。

亚细胞结构中的活性小分子对细胞功能发挥关键的调控作用，与疾病的发生发展密切相关。针对活性小分子分析中的超灵敏检测、细胞器靶向、信号时空分辨等关键科学问题，本项目基于有机合成技术和纳米生物技术，通过多重放大策略，发展了高灵敏探针，实现细胞内源性低浓度的小分子检测；通过提高探针的特异性，结合光谱可分辨，实现活性小分子原位、瞬时、动态、可逆示踪与多组分成像；最终利用探针的细胞器靶向作用，研究了亚细胞结构不同分子间相互作用及对细胞功能的影响，并通过小分子变化调控细胞命运。本项目为深入揭示亚细胞结构上活性小分子的生物学作用提供了有力的技术支撑和重要信息。本项目资助工作在国际学术期刊发表68篇论文（其中2篇J. Am. Chem. Soc.，13篇Angew. Chem. Int. Ed.，2篇Nat. Commun.），申请国家发明专利10项，5项获得授权。项目执行期间，培养博士生5人，硕士生20人。