An intact canonical NF-κB pathway is required for inflammatory gene expression in response to hypoxia.

Susan F Fitzpatrick, Murtaza Tambuwala, Ulrike Bruning, Bettina Schaible, Carsten C Scholz, Annette Byrne, Aisling O'Connor, William M Gallagher, Colin R Lenihan, John F Garvey, Katherine Howell, Padraic G Fallon, Eoin P Cummins, Cormac T Taylor

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    Abstract

    Hypoxia is a feature of the microenvironment in a number of chronic inflammatory conditions due to increased metabolic activity and disrupted perfusion at the inflamed site. Hypoxia contributes to inflammation through the regulation of gene expression via key oxygen-sensitive transcriptional regulators including the hypoxia-inducible factor (HIF) and NF-κB. Recent studies have revealed a high degree of interdependence between HIF and NF-κB signaling; however, the relative contribution of each to hypoxia-induced inflammatory gene expression remains unclear. In this study, we use transgenic mice expressing luciferase under the control of NF-κB to demonstrate that hypoxia activates NF-κB in the heart and lungs of mice in vivo. Using small interfering RNA targeted to the p65 subunit of NF-κB, we confirm a unidirectional dependence of hypoxic HIF-1α accumulation upon an intact canonical NF-κB pathway in cultured cells. Cyclooxygenase-2 and other key proinflammatory genes are transcriptionally induced by hypoxia in a manner that is both HIF-1 and NF-κB dependent, and in mouse embryonic fibroblasts lacking an intact canonical NF-κB pathway, there is a loss of hypoxia-induced inflammatory gene expression. Finally, under conditions of hypoxia, HIF-1α and the p65 subunit of NF-κB directly bind to the cyclooxygenase-2 promoter. These results implicate an essential role for NF-κB signaling in inflammatory gene expression in response to hypoxia both through the regulation of HIF-1 and through direct effects upon target gene expression.
    LanguageEnglish
    Pages1091-1096
    JournalJ Immunol
    Volume186
    Issue number2
    DOIs
    Publication statusPublished - 13 Dec 2010

    Fingerprint

    Gene Expression
    Hypoxia-Inducible Factor 1
    Cyclooxygenase 2
    Hypoxia
    Gene Expression Regulation
    Luciferases
    Small Interfering RNA
    Transgenic Mice
    Cultured Cells
    Perfusion
    Fibroblasts
    Oxygen
    Inflammation
    Lung
    Genes

    Cite this

    Fitzpatrick, S. F., Tambuwala, M., Bruning, U., Schaible, B., Scholz, C. C., Byrne, A., ... Taylor, C. T. (2010). An intact canonical NF-κB pathway is required for inflammatory gene expression in response to hypoxia. J Immunol, 186(2), 1091-1096. https://doi.org/10.4049/jimmunol.1002256
    Fitzpatrick, Susan F ; Tambuwala, Murtaza ; Bruning, Ulrike ; Schaible, Bettina ; Scholz, Carsten C ; Byrne, Annette ; O'Connor, Aisling ; Gallagher, William M ; Lenihan, Colin R ; Garvey, John F ; Howell, Katherine ; Fallon, Padraic G ; Cummins, Eoin P ; Taylor, Cormac T. / An intact canonical NF-κB pathway is required for inflammatory gene expression in response to hypoxia. In: J Immunol. 2010 ; Vol. 186, No. 2. pp. 1091-1096.
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    abstract = "Hypoxia is a feature of the microenvironment in a number of chronic inflammatory conditions due to increased metabolic activity and disrupted perfusion at the inflamed site. Hypoxia contributes to inflammation through the regulation of gene expression via key oxygen-sensitive transcriptional regulators including the hypoxia-inducible factor (HIF) and NF-κB. Recent studies have revealed a high degree of interdependence between HIF and NF-κB signaling; however, the relative contribution of each to hypoxia-induced inflammatory gene expression remains unclear. In this study, we use transgenic mice expressing luciferase under the control of NF-κB to demonstrate that hypoxia activates NF-κB in the heart and lungs of mice in vivo. Using small interfering RNA targeted to the p65 subunit of NF-κB, we confirm a unidirectional dependence of hypoxic HIF-1α accumulation upon an intact canonical NF-κB pathway in cultured cells. Cyclooxygenase-2 and other key proinflammatory genes are transcriptionally induced by hypoxia in a manner that is both HIF-1 and NF-κB dependent, and in mouse embryonic fibroblasts lacking an intact canonical NF-κB pathway, there is a loss of hypoxia-induced inflammatory gene expression. Finally, under conditions of hypoxia, HIF-1α and the p65 subunit of NF-κB directly bind to the cyclooxygenase-2 promoter. These results implicate an essential role for NF-κB signaling in inflammatory gene expression in response to hypoxia both through the regulation of HIF-1 and through direct effects upon target gene expression.",
    author = "Fitzpatrick, {Susan F} and Murtaza Tambuwala and Ulrike Bruning and Bettina Schaible and Scholz, {Carsten C} and Annette Byrne and Aisling O'Connor and Gallagher, {William M} and Lenihan, {Colin R} and Garvey, {John F} and Katherine Howell and Fallon, {Padraic G} and Cummins, {Eoin P} and Taylor, {Cormac T}",
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    Fitzpatrick, SF, Tambuwala, M, Bruning, U, Schaible, B, Scholz, CC, Byrne, A, O'Connor, A, Gallagher, WM, Lenihan, CR, Garvey, JF, Howell, K, Fallon, PG, Cummins, EP & Taylor, CT 2010, 'An intact canonical NF-κB pathway is required for inflammatory gene expression in response to hypoxia.', J Immunol, vol. 186, no. 2, pp. 1091-1096. https://doi.org/10.4049/jimmunol.1002256

    An intact canonical NF-κB pathway is required for inflammatory gene expression in response to hypoxia. / Fitzpatrick, Susan F; Tambuwala, Murtaza; Bruning, Ulrike; Schaible, Bettina; Scholz, Carsten C; Byrne, Annette; O'Connor, Aisling; Gallagher, William M; Lenihan, Colin R; Garvey, John F; Howell, Katherine; Fallon, Padraic G; Cummins, Eoin P; Taylor, Cormac T.

    In: J Immunol, Vol. 186, No. 2, 13.12.2010, p. 1091-1096.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - An intact canonical NF-κB pathway is required for inflammatory gene expression in response to hypoxia.

    AU - Fitzpatrick, Susan F

    AU - Tambuwala, Murtaza

    AU - Bruning, Ulrike

    AU - Schaible, Bettina

    AU - Scholz, Carsten C

    AU - Byrne, Annette

    AU - O'Connor, Aisling

    AU - Gallagher, William M

    AU - Lenihan, Colin R

    AU - Garvey, John F

    AU - Howell, Katherine

    AU - Fallon, Padraic G

    AU - Cummins, Eoin P

    AU - Taylor, Cormac T

    PY - 2010/12/13

    Y1 - 2010/12/13

    N2 - Hypoxia is a feature of the microenvironment in a number of chronic inflammatory conditions due to increased metabolic activity and disrupted perfusion at the inflamed site. Hypoxia contributes to inflammation through the regulation of gene expression via key oxygen-sensitive transcriptional regulators including the hypoxia-inducible factor (HIF) and NF-κB. Recent studies have revealed a high degree of interdependence between HIF and NF-κB signaling; however, the relative contribution of each to hypoxia-induced inflammatory gene expression remains unclear. In this study, we use transgenic mice expressing luciferase under the control of NF-κB to demonstrate that hypoxia activates NF-κB in the heart and lungs of mice in vivo. Using small interfering RNA targeted to the p65 subunit of NF-κB, we confirm a unidirectional dependence of hypoxic HIF-1α accumulation upon an intact canonical NF-κB pathway in cultured cells. Cyclooxygenase-2 and other key proinflammatory genes are transcriptionally induced by hypoxia in a manner that is both HIF-1 and NF-κB dependent, and in mouse embryonic fibroblasts lacking an intact canonical NF-κB pathway, there is a loss of hypoxia-induced inflammatory gene expression. Finally, under conditions of hypoxia, HIF-1α and the p65 subunit of NF-κB directly bind to the cyclooxygenase-2 promoter. These results implicate an essential role for NF-κB signaling in inflammatory gene expression in response to hypoxia both through the regulation of HIF-1 and through direct effects upon target gene expression.

    AB - Hypoxia is a feature of the microenvironment in a number of chronic inflammatory conditions due to increased metabolic activity and disrupted perfusion at the inflamed site. Hypoxia contributes to inflammation through the regulation of gene expression via key oxygen-sensitive transcriptional regulators including the hypoxia-inducible factor (HIF) and NF-κB. Recent studies have revealed a high degree of interdependence between HIF and NF-κB signaling; however, the relative contribution of each to hypoxia-induced inflammatory gene expression remains unclear. In this study, we use transgenic mice expressing luciferase under the control of NF-κB to demonstrate that hypoxia activates NF-κB in the heart and lungs of mice in vivo. Using small interfering RNA targeted to the p65 subunit of NF-κB, we confirm a unidirectional dependence of hypoxic HIF-1α accumulation upon an intact canonical NF-κB pathway in cultured cells. Cyclooxygenase-2 and other key proinflammatory genes are transcriptionally induced by hypoxia in a manner that is both HIF-1 and NF-κB dependent, and in mouse embryonic fibroblasts lacking an intact canonical NF-κB pathway, there is a loss of hypoxia-induced inflammatory gene expression. Finally, under conditions of hypoxia, HIF-1α and the p65 subunit of NF-κB directly bind to the cyclooxygenase-2 promoter. These results implicate an essential role for NF-κB signaling in inflammatory gene expression in response to hypoxia both through the regulation of HIF-1 and through direct effects upon target gene expression.

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    DO - 10.4049/jimmunol.1002256

    M3 - Article

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