Profiling communication networks of endogenous exosomes

项目说明/摘要 Exosome是分泌的小的细胞外小泡，大小为30-130 nm。它们含有DNA片段，mRNA 和蛋白质，并作为一种新的信使类别出现，促进细胞、组织和器官之间的串扰。 追踪这些细胞外小泡的分布和动态性质，并量化它们的动力学 必不可少的。迫切需要开发一种灵敏的、非侵入性的内源性跟踪系统 这样的系统将为探索外切体的正常生物学功能提供一个强有力的工具。 为了满足这样的需求，我们提出了一种独特的外显体记者鼠标，通过它来揭示交流 生理条件下器官/组织或细胞之间的内源性外切体网络和调查正常 外显体生物学。一种改良的生物发光报告将被敲入老鼠体内，这将提供一种超灵敏 和稳定的标签和跟踪系统。记者的特定诱导将在心肌细胞和 以心脏成纤维细胞为例。发光信号将被非侵入性地检测。 总而言之，这项研究将提供一种强大而通用的遗传小鼠模型来实现定位 跟踪、量化来自几乎任何组织、器官或细胞谱系的内源性外体。这个 所提出的基因小鼠优于目前的体外和体内标记。

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.