A multi-omic approach to identifying biomarkers of tissue development

组织的发育、适应和退化对个体的健康有着深远的影响。生物组织的这种适应性与特化生长因子的释放有关，特化生长因子部分地通过称为“细胞外囊泡”（EV）的细胞衍生的纳米颗粒来调节。EV含有许多生长因子和参与转录事件调节的RNA分子。这些复杂的颗粒还包含脂质的复杂排列，其可能对组织细胞形成和再生的关键细胞融合事件具有影响力但尚未确定的贡献。然而，目前对EV在组织适应、老化和退化过程中的贡献的理解仍然有限。这主要是由于（1）缺乏有效的方法来分离/纯化EV;（2）对EV的结构生化组成及其与组织发育的关系了解有限。在该项目中，我们的目标是推动EV研究的进展，以加速新医疗保健技术的发展：基于我们先前在EV分离和分析方面的工作，优化高度特异性的EV分离方案，以提高EV的检出率和特异性。回收，同时消除污染生物颗粒的存在。该方案将用于分离来自细胞培养实验的EV，以确保在项目分析阶段鉴定的生物标志物对EV具有特异性。这将确保研究结果在医疗技术领域具有转化价值。应用最先进的分析技术来阐明从细胞培养实验中分离的纯EV的蛋白质、脂质和代谢物含量。这些分析将与项目合作伙伴沃茨合作进行。我们将利用多组学方法，利用色谱法和质谱法的现代进步，对EV组成进行开创性的基于发现的蛋白质组学，脂质组学和代谢组学研究。通过在组织发育和适应期间绘制EV的内容和功能，我们提供了一种识别生物标志物的优雅方式，该生物标志物可用于预测和监测细胞适应/退化，以提供组织状态的快速和非侵入性生物标志物。此外，更深入地了解健康和疾病中EV含量的生物物理和生物化学变化将使我们能够制定在组织工程中开发EV的策略。这两个方面将为开发预测性和治疗性医疗保健技术提供基础。

Tissue development, adaptation and degeneration has a profound effect on the health of an individual. This adaptability of biological tissues is associated with the release of specialised growth factors which is partly modulated through cell-derived nanoparticles termed 'extracellular vesicles' (EVs). EVs contain numerous growth factors and RNA molecules implicated in the modulation of transcriptional events. These complex particles also contain a complex arrangement of lipids, which are likely to have influential but as yet undefined contributions to cell fusion events critical for tissue cell formation and regeneration. However, current understanding of the contribution of EVs during tissue adaptation, ageing and degeneration remains limited. This is principally due to (1) a lack of efficient methodologies for the isolation/purification of EVs and (2) a limited understanding of their structural biochemical composition and how this relates to tissue development.Within this project, we aim to drive advances in EV research to accelerate the development of new healthcare technologies by:Building on our previous work on EV isolation and analysis to optimise a highly specific EV isolation protocol that will enhance the rate and specificity of EV recovery while eliminating the presence of contaminating biological particulates. The protocol will be applied for the isolation of EVs derived from cell culture experiments to ensure that biomarkers identified in the analytical phase of the project are specific to EVs. This will ensure that outcomes generated in the study have translational value in the healthcare technologies sector.Applying state-of-the-art analytical techniques to elucidate the protein, lipid and metabolite content of pure EVs isolated from cell culture experiments. These analyses will be carried out in collaboration with the project partner, Waters. We will utilise a multi-omics approach that exploits modern advances in chromatography and mass spectrometry to perform pioneering discovery-based proteomics, lipidomics and metabolomics investigations into EV composition. By mapping the content and function of EVs during tissue development and adaptation we provide an elegant way of identifying biomarkers that can be applied for the prediction and monitoring of cellular adaptation/degeneration to deliver a rapid and non-invasive biomarker of tissue status. Furthermore, a deeper understanding of biophysical and biochemical changes in EV content in health and disease will allow us to develop strategies for the exploitation of EVs in tissue engineering These two facets will provide a foundation to develop both predictive and curative healthcare technologies.