Engineering extracellular vesicles derived from human mesenchymal stem cells

细胞外囊泡（Extracellular vesicles，EV）是一种小的、膜状的、分泌的蛋白质和核酸载体，能够实现细胞间的通讯。它们在进化上是保守的（从酵母到人类），并且在各种生物技术应用中使用EV的兴趣越来越大，特别是那些由内在再生细胞类型（如成体干细胞）分泌的EV。我们已经设计了几个永生化的人克隆间充质干细胞（MSC）系，并证明它们可以产生不同类型的EV。因此，从理论上讲，这些永生化的人类细胞系可以用来生产取之不尽的MSC-EV。然而，能够控制EV产生的效率，EV货物和靶细胞类型将具有明显的生物技术优势。该项目将建立在酵母遗传学和EV生物学的机械知识基础上，开发生物工程MSC-EV生产平台。具体目标是：-表征天然MSC-EV并优化其分离。-该项目将由Paul Genever教授和ChrisMacDonald博士共同指导，他们在MSC生物学、EV生产、干细胞和酵母模型系统、内体运输和基因工程方面具有广泛的专业知识。关键技术包括细胞培养、慢病毒转导和过表达分析、转录组学、蛋白质组学、生物信息学和EV功能的体外测定。

Engineering extracellular vesicles derived from human mesenchymal stemcellsExtracellular vesicles (EVs) are small, membranous, secreted carriers of proteinsand nucleic acids that enable cell-cell communication. They are evolutionarilyconserved (from yeast to human) and there is a growing interest in the use ofEVs in diverse biotechnological applications, particularly those secreted byintrinsically regenerative cell types such as adult stem cells. We haveengineered several immortalised human clonal mesenchymal stem cell (MSC)lines and demonstrated that they produce different types of EVs. In theorytherefore, these immortalised human cell lines may be used to produce aninexhaustible supply of MSC-EVs. However, being able to control the efficiencyof EV generation, the EV cargo and target cell type would have distinctbiotechnological advantages. This project will build on the mechanisticknowledge in yeast genetics and EV biology to develop bioengineered MSC-EVproduction platforms.Specific objectives are to:-Characterise native MSC-EVs and optimise their isolation.-Determine the underlying mechanisms to help direct and/or enrich cargo-loading in MSC-EVs-Engineer MSC-EVs to target specific cell types.The project will be co-supervised by Professor Paul Genever and Dr. ChrisMacDonald who have broad expertise in MSC biology, EV production, stem celland yeast model systems, endosomal trafficking and genetic engineering. Keytechniques will include cell culture, lentiviral transduction and over-expressionanalyses, transcriptomics, proteomics, bioinformatics and in vitro assays of EVfunction.