RNA sequencing unravels novel L cell constituents and mechanisms of GLP-1 secretion in human gastric bypass-operated intestine.

Autor: Miskelly MG; Neuroendocrine Cell Biology, Lund University Diabetes Centre, Lund University, Malmö, Sweden., Lindqvist A; Neuroendocrine Cell Biology, Lund University Diabetes Centre, Lund University, Malmö, Sweden., Piccinin E; Department of Translational Biomedicine and Neuroscience, University of Bari 'Aldo Moro', Bari, Italy.; Department of Interdisciplinary Medicine, University of Bari 'Aldo Moro', Bari, Italy., Hamilton A; Molecular Metabolism, Lund University Diabetes Centre, Lund University, Malmö, Sweden.; Islet Cell Exocytosis, Lund University Diabetes Centre, Lund University, Malmö, Sweden., Cowan E; Islet Cell Exocytosis, Lund University Diabetes Centre, Lund University, Malmö, Sweden., Nergård BJ; Aleris Obesitas, Lund, Sweden., Del Giudice R; Department of Experimental Medical Science, Lund University, Lund, Sweden.; Department of Biomedical Science and Biofilms - Research Center for Biointerfaces, Malmö University, Malmö, Sweden., Ngara M; Neuroendocrine Cell Biology, Lund University Diabetes Centre, Lund University, Malmö, Sweden., Cataldo LR; Molecular Metabolism, Lund University Diabetes Centre, Lund University, Malmö, Sweden.; Novo Nordisk Foundation Centre for Basic Metabolic Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark., Kryvokhyzha D; Bioinformatics Unit, Lund University Diabetes Centre, Lund University, Malmö, Sweden., Volkov P; Bioinformatics Unit, Lund University Diabetes Centre, Lund University, Malmö, Sweden., Engelking L; Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX, USA.; Department of Molecular Genetics, University of Texas Southwestern Medical Center, Dallas, TX, USA., Artner I; Endocrine Cell Differentiation and Function, Stem Cell Centre, Lund University, Malmö, Sweden., Lagerstedt JO; Islet Cell Exocytosis, Lund University Diabetes Centre, Lund University, Malmö, Sweden.; Department of Experimental Medical Science, Lund University, Lund, Sweden., Eliasson L; Islet Cell Exocytosis, Lund University Diabetes Centre, Lund University, Malmö, Sweden., Ahlqvist E; Genomics, Diabetes and Endocrinology, Lund University Diabetes Centre, Lund University, Malmö, Sweden., Moschetta A; Department of Interdisciplinary Medicine, University of Bari 'Aldo Moro', Bari, Italy.; INBB National Institute for Biostructure and Biosystems, Rome, Italy., Hedenbro J; Department of Surgery, Department of Clinical Sciences Lund, Lund University, Lund, Sweden., Wierup N; Neuroendocrine Cell Biology, Lund University Diabetes Centre, Lund University, Malmö, Sweden. nils.wierup@med.lu.se.
Jazyk: angličtina
Zdroj: Diabetologia [Diabetologia] 2024 Feb; Vol. 67 (2), pp. 356-370. Date of Electronic Publication: 2023 Nov 30.
DOI: 10.1007/s00125-023-06046-8
Abstrakt: Aims/hypothesis: Roux-en-Y gastric bypass surgery (RYGB) frequently results in remission of type 2 diabetes as well as exaggerated secretion of glucagon-like peptide-1 (GLP-1). Here, we assessed RYGB-induced transcriptomic alterations in the small intestine and investigated how they were related to the regulation of GLP-1 production and secretion in vitro and in vivo.
Methods: Human jejunal samples taken perisurgically and 1 year post RYGB (n=13) were analysed by RNA-seq. Guided by bioinformatics analysis we targeted four genes involved in cholesterol biosynthesis, which we confirmed to be expressed in human L cells, for potential involvement in GLP-1 regulation using siRNAs in GLUTag and STC-1 cells. Gene expression analyses, GLP-1 secretion measurements, intracellular calcium imaging and RNA-seq were performed in vitro. OGTTs were performed in C57BL/6j and iScd1 -/- mice and immunohistochemistry and gene expression analyses were performed ex vivo.
Results: Gene Ontology (GO) analysis identified cholesterol biosynthesis as being most affected by RYGB. Silencing or chemical inhibition of stearoyl-CoA desaturase 1 (SCD1), a key enzyme in the synthesis of monounsaturated fatty acids, was found to reduce Gcg expression and secretion of GLP-1 by GLUTag and STC-1 cells. Scd1 knockdown also reduced intracellular Ca 2+ signalling and membrane depolarisation. Furthermore, Scd1 mRNA expression was found to be regulated by NEFAs but not glucose. RNA-seq of SCD1 inhibitor-treated GLUTag cells identified altered expression of genes implicated in ATP generation and glycolysis. Finally, gene expression and immunohistochemical analysis of the jejunum of the intestine-specific Scd1 knockout mouse model, iScd1 -/- , revealed a twofold higher L cell density and a twofold increase in Gcg mRNA expression.
Conclusions/interpretation: RYGB caused robust alterations in the jejunal transcriptome, with genes involved in cholesterol biosynthesis being most affected. Our data highlight SCD as an RYGB-regulated L cell constituent that regulates the production and secretion of GLP-1.
(© 2023. The Author(s).)
Databáze: MEDLINE
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