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Molecular Analysis of the Diversity of Lymphoid Cell Types and Functions at Single-Cell Resolution: ProteinSimple Single-Cell Western System.

单细胞分辨率下淋巴细胞类型和功能多样性的分子分析：ProteinSimple 单细胞蛋白质印迹系统。

基本信息

批准号：
RTI-2019-00157
负责人：
Nijnik, Anastasia
金额：
$ 9.71万
依托单位：
McGill University
依托单位国家：
加拿大
项目类别：
Research Tools and Instruments
财政年份：
2018
资助国家：
加拿大
起止时间：
2018-01-01 至 2019-12-31
项目状态：
已结题

来源：
https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=660958
关键词：
Molecular Analysis Diversity Lymphoid Cell

项目摘要

Protein composition of a cell defines its functional properties. Quantitative measurements of cellular proteins are therefore the cornerstone of most research programs in life sciences. It is increasingly recognized that an accurate understanding of cellular function requires studies with primary cells from the tissues, and can not be accurately modeled with transformed cell-lines in culture or other in vitro systems. However, most primary cells are found in very low numbers in the available biological samples, and are heterogeneous in nature. This is especially true for the cells of adaptive immune system, B and T lymphocytes. Each of these cells acquires an antigen recognition receptor of a unique structure and specificity during its development, and undergoes further differentiation, polarization, and functional specification during its engagement in immune response. Precise receptor stimulation thresholds dictate cell-fate, affecting how a lymphocyte engages in immune response or enforcing cell-anergy and apoptosis. The traditional experimental methods, like Western blotting, analyze bulk cell populations and do not accurately capture the functional complexity of the cells of adaptive immune system. The molecular analysis of rare cell types at single cell resolution is therefore critical for accurate understanding of lymphocyte functions and the mechanisms of adaptive immune response and immune regulation.******This application brings together a team of outstanding immunologists from across McGill University, studying the molecular mechanisms that regulate B and T cell functions in immune response, and facing the common challenge of protein quantification in small and heterogeneous biological samples.The Milo Single-Cell Western System, requested on this application, is the only instrument of its kind on the market allowing protein quantification in small biological samples at single cell resolution, and is not currently available at McGill University and the Montreal area. The equipment is therefore urgently needed and has the potential to fundamentally advance the research programs of the applicants, providing novel insight into the molecular mechanisms of B and T lymphocyte development and adaptive immune response. Such insight can not be obtained with existing or alternative equipment and methodologies.******Importantly, the applicants have an excellent track-record in HQP training, and currently their combined research teams include over 30 trainees. Most of these trainees are working on projects that require protein quantification in small and heterogeneous cell samples, and will therefore directly and strongly benefit from the requested Milo Single-Cell Western System. The benefits to the HQPs will include the direct acquisition of new technical skills, enhanced research productivity, and increased impact of their work on the research field. ******Overall, the requested equipment offers major benefits to the academic field and HQP training.**

细胞的蛋白质组成决定了其功能特性。因此，细胞蛋白质的定量测量是生命科学中大多数研究计划的基石。越来越多的人认识到，准确理解细胞功能需要用来自组织的原代细胞进行研究，而不能用培养物或其他体外系统中的转化细胞系进行准确建模。然而，大多数原代细胞在可用的生物样品中以非常低的数量被发现，并且在性质上是异质的。对于适应性免疫系统的细胞、B和T淋巴细胞尤其如此。这些细胞中的每一个在其发育期间获得具有独特结构和特异性的抗原识别受体，并且在其参与免疫应答期间经历进一步分化、极化和功能特化。精确的受体刺激阈值决定细胞命运，影响淋巴细胞如何参与免疫反应或强制细胞无能和凋亡。传统的实验方法如Western blotting分析的是大量的细胞群体，不能准确地捕捉适应性免疫系统细胞的功能复杂性。因此，以单细胞分辨率对罕见细胞类型进行分子分析对于准确理解淋巴细胞功能以及适应性免疫应答和免疫调节机制至关重要。这项申请汇集了来自麦吉尔大学各地的优秀免疫学家团队，研究在免疫应答中调节B和T细胞功能的分子机制，面对小而异质的生物样本中蛋白质定量的共同挑战，在这项申请上要求的米洛单细胞Western系统是市场上唯一一种允许以单细胞分辨率对小生物样品进行蛋白质定量的仪器，目前麦吉尔大学和蒙特利尔地区还没有。因此，迫切需要该设备，并且该设备有可能从根本上推进申请人的研究计划，为B和T淋巴细胞发育和适应性免疫应答的分子机制提供新的见解。用现有的或替代的设备和方法无法获得这种洞察力。重要的是，申请人在HQP培训方面有着出色的记录，目前他们的联合研究团队包括30多名学员。这些学员中的大多数正在从事需要在小的异质细胞样本中进行蛋白质定量的项目，因此将直接从所要求的米洛单细胞Western系统中受益。HQP的好处将包括直接获得新的技术技能，提高研究生产力，并增加他们的工作对研究领域的影响。****** 总的来说，所要求的设备对学术领域和HQP培训具有重大益处。**