Ultra-sensitive Mass spectrometry for Precision Medicine

在过去的十年里，我们见证了细胞和发育生物学的一场革命。廉价而快速的核苷酸测序方法的出现使我们能够在常规的基础上以单细胞和空间分辨率对微小样本的RNA进行量化。公正的scRNAseq给了我们前所未有的洞察力，让我们了解细胞的异质性和内部变化如何支撑大多数生物过程。然而，mRNA的表达只是细胞状态的一个方面，就功能而言可能不是最具信息量的，因为生物分子DNA、mRNA和蛋白质之间的关系在量化时是非线性的。蛋白质的丰度，也就是基因行使其功能的分子，取决于生产和破坏之间的平衡，或者翻译和降解之间的平衡。这两个过程都不是线性依赖于信使核糖核酸的丰度，这解释了为什么没有相关性。因此，要确定细胞蛋白质组的状态，我们不能依赖于mRNA丰度作为替代，我们必须直接对蛋白质进行定量。到目前为止，基于临床质谱学的蛋白质组学领域还没有能够产生与测序方法所能做到的接近的生物学见解。然而，阻碍质谱学进展的许多限制最近已经被克服，我们建议在爱丁堡为MRC资助的同事和广泛的合作者网络建立这种能力。我们建议安装一台最先进的质谱仪和相关的液相色谱分离装置。该系统将安装在爱丁堡大学国际公认的遗传和癌症研究所(IGC)质谱学设施中。该系统将由来自IGC、炎症研究中心(CIR)和爱丁堡再生医学中心(CRM)的研究人员组成的联盟使用，以调查人类疾病中细胞和疾病的异质性。我们正在做的工作将阐明在癌症、炎症以及其他细胞应激和疾病背景下蛋白质动力学是如何调节的。重要的是，我们希望询问蛋白质组的变化如何塑造患者在正常生理和疾病期间对挑战的反应。

Over the past decade, we have witnessed a revolution in cellular and developmental biology. The advent of cheap and rapid nucleotide sequencing approaches has enabled us to quantify RNA from minute samples with single-cell and spatial resolution on a routine basis. Unbiased scRNAseq has given us unprecedented insight into how cellular heterogeneity and changes within underpin most biological processes. Nevertheless, mRNA expression is just one aspect of the cellular state, and it may not be the most informative in terms of function because the relationship between biomolecules DNA, mRNA and protein, is nonlinear when quantified. The abundance of proteins, the molecules through which genes exercise their function, is dependent on the balance between production and destruction, or translation and degradation. Neither process is linearly dependent on mRNA abundance, explaining the absence of correlation. Thus, to identify the state of the cellular proteome we cannot rely on mRNA abundances as a proxy, we must directly quantify the proteins. The field of clinical mass spectrometry-based proteomics has to date not been able to generate biological insights anywhere close to what sequencing approaches have been able to do. However, many of the limitations that have hampered the progress of mass spectrometry have recently been overcome and we propose to establish such capability in Edinburgh for MRC-funded colleagues and a broad network of collaborators. We propose to install a state-of-the-art mass spectrometer with an associated liquid chromatography separation. The system will be installed in the internationally recognised Institute of Genetics and Cancer (IGC) Mass Spectrometry Facility at the University of Edinburgh. The system will be used by a consortium of investigators from the IGC, the Centre for Inflammation Research (CIR) and Centre for Regenerative Medicine (CRM) Edinburgh to investigate cellular and disease heterogeneity in human diseases. The work we are doing will shed light on how protein dynamics are regulated during cancer, inflammation and in other cellular stress and disease contexts. Importantly, we wish to interrogate how proteome changes shape the response of the patient to challenges during normal physiology and disease.