Single-Nucleus and In Situ RNA–Sequencing Reveal Cell Topographies in the Human Pancreas

作者：Luca Tosti, Yan Hang, Olivia Debnath, Sebastian Tiesmeyer, Timo B. Trefzer, Katja Steiger, Foo Wei Ten, Soeren Lukassen, Simone Ballke, Anja A. Kühl, Simone Spieckermann, Rita Bottino, Naveed Ishaque, Wilko Weichert, Seung K. Kim, Roland Eils, Christian Conrad · 发表于：Gastroenterology · 年份：2020 · DOI：10.1053/j.gastro.2020.11.010 · 被引用次数：249 · 研究领域：Single-cell and spatial transcriptomics、Pancreatic function and diabetes、Pancreatic and Hepatic Oncology Research

Background & Aims Molecular evidence of cellular heterogeneity in the human exocrine pancreas has not been yet established because of the local concentration and cascade of hydrolytic enzymes that can rapidly degrade cells and RNA upon pancreatic resection. We sought to better understand the heterogeneity and cellular composition of the pancreas in neonates and adults in healthy and diseased conditions using single-cell sequencing approaches. Methods We innovated single-nucleus RNA-sequencing protocols and profiled more than 120,000 cells from pancreata of adult and neonatal human donors. We validated the single-nucleus findings using RNA fluorescence in situ hybridization, in situ sequencing, and computational approaches. Results We created the first comprehensive atlas of human pancreas cells including epithelial and nonepithelial constituents, and uncovered 3 distinct acinar cell types, with possible implications for homeostatic and inflammatory processes of the pancreas. The comparison with neonatal single-nucleus sequencing data showed a different cellular composition of the endocrine tissue, highlighting the tissue dynamics occurring during development. By applying spatial cartography, involving cell proximity mapping through in situ sequencing, we found evidence of specific cell type neighborhoods, dynamic topographies in the endocrine and exocrine pancreas, and principles of morphologic organization of the organ. Furthermore, similar analyses in chronic pancreatitis b...