Understanding the Role of Extracellular Vesicle/Exosome Transport in the Visual System

细胞通讯通常被理解为细胞分泌信使蛋白，即细胞因子和生长因子，它们与受体细胞上的受体结合并产生预期的效果。在过去的50年里，对疾病治疗的大量研究也一直在试图调节这些信号，抑制、改变或增加它们。最近发现了一种新的信号机制，由于其新颖性，人们对其知之甚少。除了细胞因子/生长因子外，细胞还分泌细胞外囊泡（EVs），包括外泌体和微囊泡。这些膜结合颗粒含有大量的蛋白质以及RNA （mRNA和miRNA）形式的遗传物质。虽然人们知道电动汽车的存在已经很多年了，但直到最近才有效地证明电动汽车能够将它们的货物运送到其他细胞中，而这些受体细胞能够利用这些货物。重要的是，这种mRNA形成了新蛋白质合成的蓝图，而miRNA则相反，抑制蛋白质合成。因此，细胞可以通过ev的分泌来调节基因表达，并最终调节受体细胞的蛋白质组成。由于我们对这一过程的理解不足，该项目试图更好地理解这种信号机制在眼睛正常生理中所起的作用。眼睛是一个容易接近的组织，使其成为研究EV通信的高度适用的模型。EVs与疾病有关，这包括促进发病机制和潜在治疗的有害因素。没有足够的数据表明它们在健康的身体和眼睛中的正常功能，这使得这些影响难以研究。该项目首先分离/分析ev（来自啮齿动物和人类视网膜细胞），包括它们的大小、数量和数量。这些货物包括它们的mRNA、miRNA和蛋白质。由于每个细胞释放不同类型的ev，因此确定哪些视网膜细胞释放哪些ev并跟踪它们在眼睛中的位置（包括它们的最终目的地）将非常重要。人类视网膜将从胚胎干细胞系中产生，并将作为一个有用的比较，以确保结果适用于人类生理学。我们还将确定当我们开始抑制电动汽车的释放或特定货物的包装时，眼睛会发生什么，从而使我们能够赋予电动汽车特定的功能。也有必要分析目标视网膜细胞，并更好地了解当EV运送货物时发生了什么变化。

Cellular communication has typically been understood as cells secreting messenger proteins, known as cytokines and growth factors, which bind to receptors on recipient cells and elicit their desired effect. A great deal of research over the last 50 years into treatments for disease has also been trying to modulate these signals, inhibiting, altering, or increasing them. A new signalling mechanism has recently been discovered, and due to its novelty, is still poorly understood. Along with cytokines/growth factors, cells also secrete extracellular vesicles (EVs), which include exosomes and microvesicles.These membrane-bound particles contain huge numbers of proteins as well as genetic material in the form of RNA (mRNA and miRNA). While their existence has been known for many years, only recently have EVs been effectively shown to be able to deliver their cargo into other cells, with those recipient cells able to utilize said cargo. Importantly, this mRNA forms the blueprint for new protein synthesis whereas the miRNA acts in opposition, inhibiting protein synthesis. Cells can therefore regulate the gene expression and ultimately, the protein make-up of recipient cells via the secretion of EVs.Due to our poor understanding of this process, this project seeks to better understand the role this signalling mechanism plays within the normal physiology of the eye. The eye is an easily accessible tissue, making it a highly amenable model to study EV communication. EVs have been implicated in disease and this includes detrimental factors that further the pathogenesis as well as potential therapies. Not enough data exists on their normal function within the healthy body and eye, making these implications difficult to study.This project begins by isolating/analyzing EVs (from rodent and human retinal cells) including their size, numbers and cargo. This cargo includes their mRNA, miRNA, and proteins. Since every cell releases different types of EVs, it will be important to identify which retinal cells are releasing which EVs and track where they are going in the eye including their ultimate destination. Human retina will be generated from embryonic stem cell lines and will serve as a useful comparison to ensure results are applicable to human physiology. We will also determine what happens in the eye as we begin inhibiting the release of EVs or the packaging of specific cargo, allowing us to ascribe specific functions to EVs. It is also necessary to analyse the target retinal cells and better understand what changes occur when an EV delivers its cargo.