Molecular Signaling and Dysfunction of the Human Reactive Enteric Glial Cell Phenotype: Implications for GI Infection, IBD, POI, Neurological, Motility, and GI Disorders

Andromeda Liñán-Rico, Fabio Turco, Fernando Ochoa-Cortes, Alan Harzman, Bradley J. Needleman, Razvan Arsenescu, Mahmoud Abdel-Rasoul, Paolo Fadda, Iveta Grants, Emmett Whitaker, Rosario Cuomo, Fievos L. Christofi

Research output: Contribution to journalArticle

15 Citations (Scopus)

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 2+, and purinergic signaling by Ca 2+ /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 g/mL) and interferon-γ (10 g/mL) was used to induce inflammation and study molecular signaling, flow-dependent Ca 2+ 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 2+ signaling, ATP, and mechanical/flow-dependent Ca 2+ 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 2+, 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.

Original languageEnglish (US)
Pages (from-to)1812-1834
Number of pages23
JournalInflammatory bowel diseases
Volume22
Issue number8
DOIs
StatePublished - May 31 2016
Externally publishedYes

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Gastrointestinal Motility
Neuroglia
Phenotype
Adenosine Triphosphate
Infection
Inflammation
Genes
Ileus
Inflammatory Bowel Diseases
Transcription Factor RelB
Vesicular Transport Proteins
Transcription Factor RelA
Antioxidants
Interleukin-17
Irritable Bowel Syndrome
Constipation
Interleukin-8
Interleukin-1
Caspase 3
Interleukin-4

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Gastroenterology

Cite this

Liñán-Rico, Andromeda ; Turco, Fabio ; Ochoa-Cortes, Fernando ; Harzman, Alan ; Needleman, Bradley J. ; Arsenescu, Razvan ; Abdel-Rasoul, Mahmoud ; Fadda, Paolo ; Grants, Iveta ; Whitaker, Emmett ; Cuomo, Rosario ; Christofi, Fievos L. / Molecular Signaling and Dysfunction of the Human Reactive Enteric Glial Cell Phenotype : Implications for GI Infection, IBD, POI, Neurological, Motility, and GI Disorders. In: Inflammatory bowel diseases. 2016 ; Vol. 22, No. 8. pp. 1812-1834.
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title = "Molecular Signaling and Dysfunction of the Human Reactive Enteric Glial Cell Phenotype: Implications for GI Infection, IBD, POI, Neurological, Motility, and GI Disorders",
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 2+, and purinergic signaling by Ca 2+ /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 g/mL) and interferon-γ (10 g/mL) was used to induce inflammation and study molecular signaling, flow-dependent Ca 2+ 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 2+ signaling, ATP, and mechanical/flow-dependent Ca 2+ 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 2+, 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.",
author = "Andromeda Li{\~n}{\'a}n-Rico and Fabio Turco and Fernando Ochoa-Cortes and Alan Harzman and Needleman, {Bradley J.} and Razvan Arsenescu and Mahmoud Abdel-Rasoul and Paolo Fadda and Iveta Grants and Emmett Whitaker and Rosario Cuomo and Christofi, {Fievos L.}",
year = "2016",
month = "5",
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doi = "10.1097/MIB.0000000000000854",
language = "English (US)",
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pages = "1812--1834",
journal = "Inflammatory Bowel Diseases",
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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 & Christofi, FL 2016, 'Molecular Signaling and Dysfunction of the Human Reactive Enteric Glial Cell Phenotype: Implications for GI Infection, IBD, POI, Neurological, Motility, and GI Disorders', Inflammatory bowel diseases, vol. 22, no. 8, pp. 1812-1834. https://doi.org/10.1097/MIB.0000000000000854

Molecular Signaling and Dysfunction of the Human Reactive Enteric Glial Cell Phenotype : Implications for GI Infection, IBD, POI, Neurological, Motility, and GI Disorders. / Liñán-Rico, Andromeda; Turco, Fabio; Ochoa-Cortes, Fernando; Harzman, Alan; Needleman, Bradley J.; Arsenescu, Razvan; Abdel-Rasoul, Mahmoud; Fadda, Paolo; Grants, Iveta; Whitaker, Emmett; Cuomo, Rosario; Christofi, Fievos L.

In: Inflammatory bowel diseases, Vol. 22, No. 8, 31.05.2016, p. 1812-1834.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Molecular Signaling and Dysfunction of the Human Reactive Enteric Glial Cell Phenotype

T2 - Implications for GI Infection, IBD, POI, Neurological, Motility, and GI Disorders

AU - Liñán-Rico, Andromeda

AU - Turco, Fabio

AU - Ochoa-Cortes, Fernando

AU - Harzman, Alan

AU - Needleman, Bradley J.

AU - Arsenescu, Razvan

AU - Abdel-Rasoul, Mahmoud

AU - Fadda, Paolo

AU - Grants, Iveta

AU - Whitaker, Emmett

AU - Cuomo, Rosario

AU - Christofi, Fievos L.

PY - 2016/5/31

Y1 - 2016/5/31

N2 - 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 2+, and purinergic signaling by Ca 2+ /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 g/mL) and interferon-γ (10 g/mL) was used to induce inflammation and study molecular signaling, flow-dependent Ca 2+ 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 2+ signaling, ATP, and mechanical/flow-dependent Ca 2+ 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 2+, 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.

AB - 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 2+, and purinergic signaling by Ca 2+ /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 g/mL) and interferon-γ (10 g/mL) was used to induce inflammation and study molecular signaling, flow-dependent Ca 2+ 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 2+ signaling, ATP, and mechanical/flow-dependent Ca 2+ 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 2+, 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.

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