TY - JOUR
T1 - High-Resolution mRNA and Secretome Atlas of Human Enteroendocrine Cells
AU - Beumer, Joep
AU - Puschhof, Jens
AU - Bauza-Martinez, Julia
AU - Martinez-Silgado, Adriana
AU - Elmentaite, Rasa
AU - James, Kylie R.
AU - Ross, Alexander
AU - Hendriks, Delilah
AU - Artegiani, Benedetta
AU - Busslinger, Georg A.
AU - Ponsioen, Bas
AU - Andersson-Rolf, Amanda
AU - Saftien, Aurelia
AU - Boot, Charelle
AU - Kretzschmar, Kai
AU - Geurts, Maarten H.
AU - Bar-Ephraim, Yotam E.
AU - Pleguezuelos-Manzano, Cayetano
AU - Post, Yorick
AU - Begthel, Harry
AU - van der Linden, Franka
AU - Lopez-Iglesias, Carmen
AU - van de Wetering, Willine J.
AU - van der Linden, Reinier
AU - Peters, Peter J.
AU - Heck, Albert J. R.
AU - Goedhart, Joachim
AU - Snippert, Hugo
AU - Zilbauer, Matthias
AU - Teichmann, Sarah A.
AU - Wu, Wei
AU - Clevers, Hans
N1 - Funding Information:
We thank Anko de Graaff and the Hubrecht Imaging Centre (HIC) for microscopy assistance, Single Cell Discoveries for single-cell sequencing, and Folkert Morsink and Johan Offerhaus of the University Medical Center Utrecht for providing sections of human intestinal biopsies. J.B.-M. A.J.R.H. and W.W. acknowledge financial support from the Horizon 2020 program INFRAIA project Epic-XS (Project 823839) and the NWO funded Netherlands Proteomics Centre through the National Road Map for Large-scale Infrastructures program X-Omics (Project 184.034.019). This work was supported by NETRF/Petersen Accelerator (J.B.), CRUK grant OPTIMISTICC (C10674/A27140) (J.P. and C.P.-M.), Netherlands Organ-on-Chip Initiative (024.003.001) from the Netherlands Organisation for Scientific Research (NWO) (J.P. A.M.-S. and C.P.-M.), and ERC Advanced Grant Agreement no. 67013e (H.C.). We thank the Microscopy CORE Lab at M4I Maastricht University for their support in electron microscopy. J.B. J.P. and H.C. conceptualized the project, designed the experiments, interpreted the results, and wrote the manuscript. J.B.-M. W.W. and A.J.R.H. performed the proteomic and secretomic experiments and analysis. A.M.S. assisted in cell culture experiments supervised by J.B. J.P. and H.C.; J.B. and J.P generated and analyzed the organoid-derived EEC single-cell atlas. R.E. K.R.J. A.R. M.Z. and S.A.T. generated and analyzed the primary human EEC single-cell dataset. G.A.B. generated and provided the neurogenin-3 overexpression vector. D.H. and B.A. generated and provided the targeting vectors for CRISPR-HOT-mediated reporter organoids. A.A.-R. and M.H.G provided different constructs. A.S. C.B. Y.P. and C.P.-M. contributed to functional EEC assays. F.v.d.L. and J.G. provided the Tq-Ca-FLITS reporter. B.P. and H.S. performed imaging and quantification of calcium responses. Y.E.B.-E. and R.v.d.L assisted with FACS experiments. K.K. provided organoid lines. C.L.-I. W.J.v.d.W. and P.J.P. performed transmission electron microscopy. H.C. is inventor on several patents related to organoid technology; his full disclosure is given at https://www.uu.nl/staff/JCClevers/. H.C. is founder of OrganoidZ, which employs organoids for drug development. J.B. J.P. and H.C. are inventors on patents related to this work.
Publisher Copyright:
© 2020 Elsevier Inc.
PY - 2020/6/11
Y1 - 2020/6/11
N2 - Enteroendocrine cells (EECs) sense intestinal content and release hormones to regulate gastrointestinal activity, systemic metabolism, and food intake. Little is known about the molecular make-up of human EEC subtypes and the regulated secretion of individual hormones. Here, we describe an organoid-based platform for functional studies of human EECs. EEC formation is induced in vitro by transient expression of NEUROG3. A set of gut organoids was engineered in which the major hormones are fluorescently tagged. A single-cell mRNA atlas was generated for the different EEC subtypes, and their secreted products were recorded by mass-spectrometry. We note key differences to murine EECs, including hormones, sensory receptors, and transcription factors. Notably, several hormone-like molecules were identified. Inter-EEC communication is exemplified by secretin-induced GLP-1 secretion. Indeed, individual EEC subtypes carry receptors for various EEC hormones. This study provides a rich resource to study human EEC development and function.
AB - Enteroendocrine cells (EECs) sense intestinal content and release hormones to regulate gastrointestinal activity, systemic metabolism, and food intake. Little is known about the molecular make-up of human EEC subtypes and the regulated secretion of individual hormones. Here, we describe an organoid-based platform for functional studies of human EECs. EEC formation is induced in vitro by transient expression of NEUROG3. A set of gut organoids was engineered in which the major hormones are fluorescently tagged. A single-cell mRNA atlas was generated for the different EEC subtypes, and their secreted products were recorded by mass-spectrometry. We note key differences to murine EECs, including hormones, sensory receptors, and transcription factors. Notably, several hormone-like molecules were identified. Inter-EEC communication is exemplified by secretin-induced GLP-1 secretion. Indeed, individual EEC subtypes carry receptors for various EEC hormones. This study provides a rich resource to study human EEC development and function.
KW - GLUCAGON-LIKE PEPTIDE-1
KW - SMALL-INTESTINE
KW - CHROMOGRANIN-A
KW - STEM-CELLS
KW - GENE
KW - RECEPTOR
KW - PROTEIN
KW - SEROTONIN
KW - DIFFERENTIATION
KW - EXPRESSION
U2 - 10.1016/j.cell.2020.04.036
DO - 10.1016/j.cell.2020.04.036
M3 - Article
C2 - 32407674
SN - 0092-8674
VL - 181
SP - 1291-1306.e19
JO - Cell
JF - Cell
IS - 6
ER -