Table_1_Near-Single-Cell Proteomics Profiling of the Proximal Tubular and Glomerulus of the Normal Human Kidney.xlsx

Molecular assessments at the single cell level can accelerate biological research by providing detailed assessments of cellular organization and tissue heterogeneity in both disease and health. The human kidney has complex multi-cellular states with varying functionality, much of which can now be completely harnessed with recent technological advances in tissue proteomics at a near single-cell level. We discuss the foundational steps in the first application of this mass spectrometry (MS) based proteomics method for analysis of sub-sections of the normal human kidney, as part of the Kidney Precision Medicine Project (KPMP). Using ~30–40 laser captured micro-dissected kidney cells, we identified more than 2,500 human proteins, with specificity to the proximal tubular (PT; n = 25 proteins) and glomerular (Glom; n = 67 proteins) regions of the kidney and their unique metabolic functions. This pilot study provides the roadmap for application of our near-single-cell proteomics workflow for analysis of other renal micro-compartments, on a larger scale, to unravel perturbations of renal sub-cellular function in the normal kidney as well as different etiologies of acute and chronic kidney disease.单细胞水平的分子表征可通过对疾病与健康状态下的细胞组织结构及组织异质性进行精细化解析，加速生物学研究进程。人体肾脏存在功能各异的复杂多细胞状态，而近年来近乎单细胞分辨率的组织蛋白质组学技术突破，使得我们得以充分挖掘其中蕴含的海量信息。作为肾脏精准医学项目（Kidney Precision Medicine Project, KPMP）的组成部分，我们详述了基于质谱（mass spectrometry, MS）的蛋白质组学方法首次应用于正常人体肾脏亚区域分析的基础流程步骤。本研究通过对约30至40个激光捕获显微切割的肾脏细胞进行分析，成功鉴定出超过2500个人类蛋白质，其中包括25种特异性表达于近端肾小管（proximal tubular, PT；n=25）区域、67种特异性表达于肾小球（glomerular, Glom；n=67）区域的蛋白质，以及这些区域特有的代谢功能。本先导研究为规模化应用近乎单细胞分辨率的蛋白质组学流程分析其他肾脏微区室提供了实践指南，助力解析正常肾脏以及急性、慢性肾脏病不同病因下的肾脏亚细胞功能扰动。