Mechanisms of extracellular vesicle biogenesis that regulate wound healing

摘要 皮肤伤口愈合中的解决方案是通过细胞间通讯过程协调的， 包括含有生物活性蛋白质和核酸的胞外小泡(EV)。在受伤中 模型，EVS已被证明是组织修复的重要媒介，尽管 特定电动汽车子集形成的潜在机制，电动汽车有效载荷的加载，以及细胞- 特定类型的EV摄取仍然知之甚少。我们通过以下方式解决此知识差距 EV生物发生调控改变EV的综合遗传方法 具有明确生物学特征的皮肤创伤模型中的轮廓、有效载荷和活性 关联性。我们最近证实，调控EV生物发生的遗传工具是“重新编程”的。 电动汽车有效载荷。此外，已确定的特定修复有效载荷的过度表达 通过质谱学可以被设计和提供，以提高模型的分辨率 伤口愈合受阻。专注于发现新的EV串扰介体 在伤口床上的免疫细胞和上皮细胞之间，我们专注于EVS的生物学 EVS形成中的细胞间信号转导(项目1-EV的生物发生)，工程 生物活性有效载荷(项目2-电动汽车有效载荷)，并确定特定的细胞类型 使EV内化于伤口床(项目3-EV摄取)。这些项目共同解决了 总体目标是用系统化的方法创造治疗性电动汽车， 基于对1)电动汽车是如何以及在哪里制造的理解的特定细胞类型；2)如何制造电动汽车 以及它们是否具有生物活性？以及3)促进哪些细胞摄取EV 持久的组织修复。

SUMMARY Resolution in skin wound healing is coordinated by intercellular communication processes that include extracellular vesicles (EVs) containing biologically active protein and nucleic acid. In injury models, EVs have been shown to be important mediators of tissue repair, although the mechanisms underlying the formation of specific EV subsets, loading of EV payloads, and cell- type specific EV uptake remain poorly understood. We address this knowledge gap with a comprehensive genetic approach to modify the regulation of EV biogenesis to change the EV Profile, Payload and Activity in cutaneous wound models that have well-defined biological relevance. We have recently established that genetic tools regulating EV biogenesis `re-program' EV payload. Moreover, over-expression of specific pro-reparative payloads that were identified by mass spectrometry can be engineered and delivered to promote resolution in models of impaired wound healing. With a focus on the discovery of novel EV mediators of crosstalk between immune and epithelial cells in the wound bed, we have focused on the biology of EVs in mediating intercellular signaling in EVs formation (Project 1-EV Biogenesis), the engineering of biologically active payloads (Project 2-EV Payloads), and identifying specific cell types that internalize EVs in the wound bed (Project 3-EV Uptake). Together, these Projects address the over-arching goals of creating therapeutic EVs with a systematic approach that is optimized for specific cell types based on an understanding of 1) How and where EVs are made?; 2) How EVs are loaded and whether they are biologically active ?; and 3) Which cells uptake EVs to promote durable tissue repair.