Profiling communication networks of endogenous exosomes

项目描述/摘要 外泌体是一种分泌型的细胞外小泡，大小为30- 130 nm。它们含有DNA片段，mRNA 和蛋白质，并作为一类新的信使出现，促进细胞，组织和器官之间的相互作用。 跟踪这些小的细胞外囊泡的分布和动态性质，并量化它们的动力学， 具有本质意义一个关键的需求出现了发展一个敏感的和非侵入性的跟踪系统，为内源性 外泌体;这样的系统将为探索外泌体的正常生物学功能提供有力的工具。 为了满足这种需求，提出了一种独特的外泌体报告小鼠，通过该小鼠来揭示通信 在生理条件下的器官/组织或细胞之间的内源性外泌体网络，并研究正常的 外泌体生物学修饰后的生物发光报告基因将被敲入小鼠体内，这将提供超灵敏的 和稳定的标签和跟踪系统。将在心肌细胞中测试报告基因的特异性诱导， 心脏成纤维细胞作为两个例子。发光信号将被非侵入性地检测。 总之，这项研究将提供一个强大的和通用的遗传小鼠模型，使定位 追踪、定量来自几乎任何组织、器官或细胞谱系的内源性外泌体。的 所提出的遗传小鼠上级目前的离体和体内标记。

PROJECT DESCRIPTION/ABSTRACT Exosomes are secreted small extracellular vesicles with the size 30-130nm. They contain DNA fragments, mRNAs and proteins, and emerge as a new category of messengers that facilitate cross-talk among cells, tissues, and organs. Tracking the distribution and dynamic nature of these small extracellular vesicles, and quantifying their kinetics are essential. A critical demand arises for the development of a sensitive and noninvasive tracking system for endogenous exosomes; such a system will provide a powerful tool for the exploration of the normal biological functions of exosomes. To meet such as a demand, a unique exosome reporter mouse is proposed, through which to reveal communication networks of endogenous exosomes among organs/tissues or cells in physiological conditions and investigate normal exosome biology. A modified bioluminescent reporter will be knocked in the mice, which will provide an ultra-sensitive and stable labeling and tracking system. The specific inductions of the reporter will be tested in cardiomyocytes and cardiac fibroblasts as two examples. The luminescent signaling will be detected non-invasively. In summary, the study will provide a powerful and versatile genetic mouse model to enable location tracking, quantifying endogenous exosomes derived from almost any tissues, organs or cell lineages. The proposed genetic mice are superior to the current ex vivo and in vivo labeling.