Ultra-sensitive Mass spectrometry for Precision Medicine

在过去的十年里，我们目睹了细胞和发育生物学的革命。廉价和快速的核苷酸测序方法的出现使我们能够在常规基础上以单细胞和空间分辨率定量来自微小样品的RNA。无偏见的scRNAseq为我们提供了前所未有的洞察力，了解细胞异质性和内部变化如何支撑大多数生物过程。然而，mRNA表达只是细胞状态的一个方面，并且它在功能方面可能不是最具信息性的，因为生物分子DNA、mRNA和蛋白质之间的关系在定量时是非线性的。蛋白质是基因发挥功能的分子，其丰度取决于产生和破坏或翻译和降解之间的平衡。这两个过程都不是线性依赖于mRNA丰度，解释了相关性的缺乏。因此，为了确定细胞蛋白质组的状态，我们不能依赖mRNA丰度作为替代，我们必须直接定量蛋白质。迄今为止，基于临床质谱的蛋白质组学领域还无法产生与测序方法所能产生的生物学见解接近的生物学见解。然而，许多限制，阻碍了质谱法的进展最近已经克服，我们建议建立这样的能力在爱丁堡的MRC资助的同事和广泛的合作者网络。我们建议安装一个国家的最先进的质谱仪与相关的液相色谱分离。该系统将安装在爱丁堡大学国际公认的遗传学和癌症研究所（IGC）质谱设施中。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用于研究人类疾病中的细胞和疾病异质性。我们正在做的工作将揭示蛋白质动力学在癌症，炎症和其他细胞应激和疾病背景下如何调节。重要的是，我们希望询问蛋白质组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.