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.

在单细胞层面上进行分子评估，能够加速生物学研究，通过提供疾病与健康状态下细胞组织结构和组织异质性的详细评估。人类肾脏拥有复杂的多元细胞状态，其功能各异，得益于近年来组织蛋白质组学在近乎单细胞水平上的技术进步，这些功能大多现已得到充分利用。本文讨论了首次将基于质谱（MS）的蛋白质组学方法应用于分析正常人类肾脏亚部分区域的奠基性步骤，作为肾脏精准医学计划（KPMP）的一部分。通过约30-40个激光捕获的肾脏细胞微切片，我们鉴定出超过2500种人类蛋白，这些蛋白特异性地分布在肾脏近端肾小管（PT；n = 25蛋白）和肾小球（Glom；n = 67蛋白）区域，并具有独特的代谢功能。这项试点研究为我们应用近乎单细胞蛋白质组学工作流程，在更大规模上分析其他肾脏微区室，揭示正常肾脏亚细胞功能的紊乱以及急性与慢性肾脏疾病不同病因提供了路线图。