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

细胞通信通常被理解为细胞分泌信使蛋白，称为细胞因子和生长因子，其结合受体细胞上的受体并引起其期望的效果。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，由于其新奇，人们对它的了解仍然很少。细胞沿着细胞因子/生长因子，还分泌细胞外囊泡（EV），包括外泌体和微囊泡，这些膜结合颗粒含有大量蛋白质以及RNA（mRNA和miRNA）形式的遗传物质。虽然它们的存在已经知道了很多年，但直到最近才有效地证明EV能够将其货物输送到其他细胞中，而那些受体细胞能够利用所述货物。重要的是，这种mRNA形成了新蛋白质合成的蓝图，而miRNA起相反的作用，抑制蛋白质合成。因此，细胞可以调节基因表达，并最终通过分泌EV调节受体细胞的蛋白质组成。由于我们对这一过程的了解不多，本项目旨在更好地了解这种信号传导机制在眼睛正常生理中的作用。眼睛是一种容易接近的组织，使其成为研究EV通信的高度可靠的模型。EV与疾病有关，这包括促进发病机制和潜在治疗的有害因素。由于没有足够的数据来说明它们在健康身体和眼睛中的正常功能，因此很难研究这些影响。该项目首先从分离/分析EV（来自啮齿动物和人类视网膜细胞），包括它们的大小，数量和货物。这种货物包括它们的mRNA，miRNA和蛋白质。由于每个细胞都会释放不同类型的EV，因此识别哪些视网膜细胞正在释放哪些EV并跟踪它们在眼睛中的位置（包括最终目的地）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，眼睛中会发生什么，使我们能够将特定功能归于电动汽车。还需要分析目标视网膜细胞，并更好地了解电动汽车运送货物时发生的变化。

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.