Table_3_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.

在单细胞层面上对分子进行评估，能够显著加速生物学研究，因为它提供了关于疾病与健康状态下细胞组织结构及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，许多功能如今可通过近期在组织蛋白质组学方面的技术进步，在近乎单细胞水平上得到完全的利用。本文探讨了首次将基于质谱（MS）的蛋白质组学方法应用于分析正常人类肾脏亚区的基础步骤，这构成了肾脏精准医学项目（KPMP）的一部分。通过使用约30-40个激光捕获的肾脏细胞微切片，我们鉴定出了超过2500种人类蛋白质，并针对肾脏近端肾小管（PT；n = 25种蛋白质）和肾小球（Glom；n = 67种蛋白质）区域及其独特的代谢功能进行了特异性分析。这一初步研究为我们提供了应用近乎单细胞蛋白质组学工作流程分析其他肾脏微区室的路线图，以更大规模地揭示正常肾脏中肾亚细胞功能的紊乱以及急性与慢性肾脏病不同病因的奥秘。