1. Molecular Signaling and Dysfunction of the Human Reactive Enteric Glial Cell Phenotype: Implications for GI Infection, IBD, POI, Neurological, Motility, and GI Disorders.
- Author
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Liñán-Rico A, Turco F, Ochoa-Cortes F, Harzman A, Needleman BJ, Arsenescu R, Abdel-Rasoul M, Fadda P, Grants I, Whitaker E, Cuomo R, and Christofi FL
- Subjects
- Adenosine Triphosphate metabolism, Calcium Channels genetics, Carrier Proteins genetics, Caspase 3 genetics, Cells, Cultured, Colon, Sigmoid cytology, Cytokines genetics, Cytokines metabolism, Enteric Nervous System cytology, Gastrointestinal Motility, Granulocyte-Macrophage Colony-Stimulating Factor genetics, Heme Oxygenase-1 genetics, Humans, Intercellular Signaling Peptides and Proteins genetics, Interferon-gamma pharmacology, Jejunum cytology, Lipopolysaccharides pharmacology, Mechanotransduction, Cellular genetics, Nitric Oxide Synthase Type II genetics, Phenotype, Receptors, Purinergic metabolism, S100 Calcium Binding Protein beta Subunit metabolism, Superoxide Dismutase genetics, Transcription Factors genetics, Tryptophan Hydroxylase genetics, Up-Regulation drug effects, Vesicular Transport Proteins genetics, Calcium metabolism, Gastrointestinal Diseases genetics, Gastrointestinal Diseases metabolism, Gene Expression drug effects, Inflammation genetics, Inflammation metabolism, Neuroglia metabolism, Receptors, Purinergic genetics, Signal Transduction genetics
- Abstract
Background: Clinical observations or animal studies implicate enteric glial cells in motility disorders, irritable bowel syndrome, inflammatory bowel disease, gastrointestinal (GI) infections, postoperative ileus, and slow transit constipation. Mechanisms underlying glial responses to inflammation in human GI tract are not understood. Our goal was to identify the "reactive human enteric glial cell (rhEGC) phenotype" induced by inflammation, and probe its functional relevance., Methods: Human enteric glial cells in culture from 15 GI-surgical specimens were used to study gene expression, Ca, and purinergic signaling by Ca/fluo-4 imaging and mechanosensitivity. A nanostring panel of 107 genes was designed as a read out of inflammation, transcription, purinergic signaling, vesicular transport protein, channel, antioxidant, and other pathways. A 24-hour treatment with lipopolysaccharide (200 μg/mL) and interferon-γ (10 μg/mL) was used to induce inflammation and study molecular signaling, flow-dependent Ca responses from 3 mL/min to 10 mL/min, adenosine triphosphate (ATP) release, and ATP responses., Results: Treatment induced a "rhEGC phenotype" and caused up-regulation in messenger RNA transcripts of 58% of 107 genes analyzed. Regulated genes included inflammatory genes (54%/IP10; IFN-γ; CxCl2; CCL3; CCL2; C3; s100B; IL-1β; IL-2R; TNF-α; IL-4; IL-6; IL-8; IL-10; IL-12A; IL-17A; IL-22; and IL-33), purine-genes (52%/AdoR2A; AdoR2B; P2RY1; P2RY2; P2RY6; P2RX3; P2RX7; AMPD3; ENTPD2; ENTPD3; and NADSYN1), channels (40%/Panx1; CHRNA7; TRPV1; and TRPA1), vesicular transporters (SYT1, SYT2, SNAP25, and SYP), transcription factors (relA/relB, SOCS3, STAT3, GATA_3, and FOXP3), growth factors (IGFBP5 and GMCSF), antioxidant genes (SOD2 and HMOX1), and enzymes (NOS2; TPH2; and CASP3) (P < 0.0001). Treatment disrupted Ca signaling, ATP, and mechanical/flow-dependent Ca responses in human enteric glial cells. ATP release increased 5-fold and s100B decreased 33%., Conclusions: The "rhEGC phenotype" is identified by a complex cascade of pro-inflammatory pathways leading to alterations of important molecular and functional signaling pathways (Ca, purinergic, and mechanosensory) that could disrupt GI motility. Inflammation induced a "purinergic switch" from ATP to adenosine diphosphate/adenosine/uridine triphosphate signaling. Findings have implications for GI infection, inflammatory bowel disease, postoperative ileus, motility, and GI disorders., Competing Interests: and The authors have declared that no conflict of interest exists.
- Published
- 2016
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