A cellenONE BSC to enhance UK capability for single-cell proteomics

cellenONE BSC 可增强英国单细胞蛋白质组学能力

• 批准号: BB/W019744/1
• 负责人: Edward Emmott
• 金额: $ 23.23万
• 依托单位: University of Liverpool
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW019744%2F1
• 关键词: cellenONE; BSC; enhance; UK; capability

Single-cell studies are transforming many areas of biological research; these include studies of infection, differentiation, construction of cell atlases, and more. Single-cell proteomics is a rapidly emerging addition to the single-cell toolset, with single-cell proteomics by mass spectrometry capable of measuring the abundance of thousands of proteins across hundreds to thousands of single-cells. By acquiring state-of-the-art instrumentation for single-cell proteomics sample preparation, this proposal seeks to enhance UK capabilities in single-cell proteomics. Specific applications of this instrument include:i) Further development and application of single-cell proteomics. The cellenONE increases the sensitivity of single-cell proteomics by minimizing sample losses during processing. We will exploit this increased sensitivity to target specific proteins of interest for reproducible analysis across single-cells, study post-translational modifications at single-cell level, as well as apply this system to develop new methods for spatial and subcellular single-cell 'omics.ii) Cell/picolitre dispensing for other single-cell approaches. In this proposal we specifically apply this instrument to single-cell western blotting, providing information on protein molecular weight, and more sensitive comparative analysis as the direct introduction of cells into nanowells permits multiple samples to be analysed per chip, minimizing batch effects. Similarly, the cellenONE instrument will be applied to single-cell RNAseq studies, where the gentle acoustic dispensing used by the system can overcome cell-size-based biases or problems analysing fragile cells in existing approaches.We propose to bring this novel instrumentation which enables automation of the entire single-cell proteomics workflow to the Centre for Proteome Research (CPR) at the University of Liverpool (UoL). Making full use of our existing suite of LC-MS/MS instruments, and using our established cost recovery model which ensures long-term maintenance and support (supported by LIV-SRF which oversees research facilities at UoL), this instrumentation will be made available to researchers working across BBSRC priority areas, both locally and nationally. LIV-SRF provides financial support to maintain and develop open facilities such as the CPR, including long-term support of staff. LIV-SRF also awards 'LIV-SRF Vouchers' which provide academics, especially ECRs with matched funds to support facility usage and develop preliminary data, to support future funding or fellowship applications. All five current cellenONE systems in academic labs in the UK are located in London or the East of England, and none are currently used for single-cell proteomics. This instrument will be applied to multiple BBSRC-funded research areas, and our demonstration projects and letters of support for this system have focused on research impacting infection biology, fundamental cell signaling research, and immunity. This instrument will enable us to both directly contribute to these research areas, as well as support training for other UK academics and industry users to establish single-cell proteomics at their own institutions.

单细胞研究正在改变生物学研究的许多领域;这些研究包括感染，分化，细胞图谱的构建等。单细胞蛋白质组学是单细胞工具集的一个快速新兴的补充，单细胞蛋白质组学通过质谱能够测量数百至数千个单细胞中数千种蛋白质的丰度。通过获得最先进的单细胞蛋白质组学样品制备仪器，该提案旨在提高英国在单细胞蛋白质组学方面的能力。该仪器的具体应用包括：i）单细胞蛋白质组学的进一步发展和应用。cellenONE通过最大限度地减少处理过程中的样品损失来提高单细胞蛋白质组学的灵敏度。我们将利用这种对目标特异性蛋白质的增加的灵敏度用于跨单细胞的可重复分析，研究单细胞水平的翻译后修饰，以及应用该系统开发用于空间和亚细胞单细胞组学的新方法。ii）用于其他单细胞方法的细胞/皮升分配。在这项提案中，我们专门将该仪器应用于单细胞蛋白质印迹，提供有关蛋白质分子量的信息，以及更灵敏的比较分析，因为直接将细胞引入细胞中允许每个芯片分析多个样品，最大限度地减少批次效应。同样，cellenONE仪器将应用于单细胞RNAseq研究，系统使用的温和声学分配可以克服现有方法中基于细胞大小的偏差或分析脆弱细胞的问题。我们建议将这种新型仪器带到利物浦大学（UoL）的蛋白质组研究中心（CPR），使整个单细胞蛋白质组学工作流程自动化。充分利用我们现有的LC-MS/MS仪器套件，并使用我们建立的成本回收模型，确保长期维护和支持（由监督UoL研究设施的LIV-SRF支持），该仪器将提供给在BBSRC优先领域工作的研究人员，无论是本地还是全国。LIV-SRF提供财政支持，以维持和发展CPR等开放式设施，包括长期支持工作人员。LIV-SRF还颁发“LIV-SRF代金券”，为学者（尤其是ECR）提供配套资金，以支持设施使用和开发初步数据，以支持未来的资助或奖学金申请。目前英国学术实验室的所有五个cellenONE系统都位于伦敦或英格兰东部，目前没有一个用于单细胞蛋白质组学。该仪器将应用于多个BBSRC资助的研究领域，我们的示范项目和支持该系统的信件集中在影响感染生物学，基础细胞信号研究和免疫力的研究上。该仪器将使我们能够直接为这些研究领域做出贡献，并支持其他英国学者和行业用户在自己的机构建立单细胞蛋白质组学的培训。