Marie-Louise Frémond1, Carolina Uggenti2, Lien Van Eyck2, Isabelle Melki3, Vincent Bondet4, Naoki Kitabayashi2, Christina Hertel5, Adrian Hayday5, Bénédicte Neven6, Yoann Rose2, Darragh Duffy7, Yanick J Crow8, Mathieu P Rodero2. 1. Paris Descartes University, Sorbonne-Paris-Cité, Institut Imagine, INSERM UMR 1163, Laboratory of Neurogenetics and Neuroinflammation, and Pediatric Hematology-Immunology and Rheumatology Department, Hôpital Necker-Enfants Malades, AP-HP, Paris, France. 2. Paris Descartes University, Sorbonne-Paris-Cité, Institut Imagine and INSERM UMR 1163, Laboratory of Neurogenetics and Neuroinflammation, Paris, France. 3. Paris Descartes University, Sorbonne-Paris-Cité, Institut Imagine, INSERM UMR 1163, Laboratory of Neurogenetics and Neuroinflammation, Pediatric Hematology-Immunology and Rheumatology Department, Hôpital Necker-Enfants Malades, AP-HP, and General Pediatrics, Infectious Diseases and Internal Medicine Department, Hôpital Robert Debré, AP-HP, Paris, France. 4. INSERM UMR 1223 and Laboratory of Dendritic Cell Immunobiology, Institut Pasteur, Paris, France. 5. ImmunoQure, Du¨sseldorf, Germany. 6. Paris Descartes University, Sorbonne-Paris-Cité, Institut Imagine, Pediatric Hematology-Immunology and Rheumatology Department, Hôpital Necker-Enfants Malades, AP-HP, and INSERM UMR 1163, Laboratory of Immunogenetics of Pediatric Autoimmune Diseases, Paris, France. 7. Darragh Duffy, PhD: INSERM UMR 1223, and Laboratory of Dendritic Cell Immunobiology, Institut Pasteur, and Center for Translational Research, Institut Pasteur, Paris, France. 8. Paris Descartes University, Sorbonne-Paris-Cité, Institut Imagine, INSERM UMR 1163, Laboratory of Neurogenetics and Neuroinflammation, Paris, France, and University of Manchester, Manchester, UK.
Abstract
OBJECTIVE: Gain-of-function mutations in TMEM173, encoding the stimulator of interferon (IFN) genes (STING) protein, underlie a novel type I interferonopathy that is minimally responsive to conventional immunosuppressive therapies and associated with high frequency of childhood morbidity and mortality. STING gain-of-function causes constitutive oversecretion of IFN. This study was undertaken to determine the effects of a TANK-binding kinase 1 (TBK-1)/IKKɛ inhibitor (BX795) on secretion and signaling of IFN in primary peripheral blood mononuclear cells (PBMCs) from patients with mutations in STING. METHODS: PBMCs from 4 patients with STING-associated disease were treated with BX795. The effect of BX795 on IFN pathways was assessed by Western blotting and an IFNβ reporter assay, as well as by quantification of IFNα in cell lysates, staining for STAT-1 phosphorylation, and measurement of IFN-stimulated gene (ISG) messenger RNA (mRNA) expression. RESULTS: Treatment of PBMCs with BX795 inhibited the phosphorylation of IFN regulatory factor 3 and IFNβ promoter activity induced in HEK 293T cells by cyclic GMP-AMP or by genetic activation of STING. In vitro exposure to BX795 inhibited IFNα production in PBMCs of patients with STING-associated disease without affecting cell survival. In addition, BX795 decreased STAT-1 phosphorylation and ISG mRNA expression independent of IFNα blockade. CONCLUSION: These findings demonstrate the effect of BX795 on reducing type I IFN production and IFN signaling in cells from patients with gain-of-function mutations in STING. A combined inhibition of TBK-1 and IKKɛ therefore holds potential for the treatment of patients carrying STING mutations, and may also be relevant in other type I interferonopathies.
OBJECTIVE: Gain-of-function mutations in TMEM173, encoding the stimulator of interferon (IFN) genes (STING) protein, underlie a novel type I interferonopathy that is minimally responsive to conventional immunosuppressive therapies and associated with high frequency of childhood morbidity and mortality. STING gain-of-function causes constitutive oversecretion of IFN. This study was undertaken to determine the effects of a TANK-binding kinase 1 (TBK-1)/IKKɛ inhibitor (BX795) on secretion and signaling of IFN in primary peripheral blood mononuclear cells (PBMCs) from patients with mutations in STING. METHODS: PBMCs from 4 patients with STING-associated disease were treated with BX795. The effect of BX795 on IFN pathways was assessed by Western blotting and an IFNβ reporter assay, as well as by quantification of IFNα in cell lysates, staining for STAT-1 phosphorylation, and measurement of IFN-stimulated gene (ISG) messenger RNA (mRNA) expression. RESULTS: Treatment of PBMCs with BX795 inhibited the phosphorylation of IFN regulatory factor 3 and IFNβ promoter activity induced in HEK 293T cells by cyclic GMP-AMP or by genetic activation of STING. In vitro exposure to BX795 inhibited IFNα production in PBMCs of patients with STING-associated disease without affecting cell survival. In addition, BX795 decreased STAT-1 phosphorylation and ISG mRNA expression independent of IFNα blockade. CONCLUSION: These findings demonstrate the effect of BX795 on reducing type I IFN production and IFN signaling in cells from patients with gain-of-function mutations in STING. A combined inhibition of TBK-1 and IKKɛ therefore holds potential for the treatment of patients carrying STING mutations, and may also be relevant in other type I interferonopathies.
Authors: M Javad Wahadat; Iris L A Bodewes; Naomi I Maria; Cornelia G van Helden-Meeuwsen; Annette van Dijk-Hummelman; Eline C Steenwijk; Sylvia Kamphuis; Marjan A Versnel Journal: Arthritis Res Ther Date: 2018-01-10 Impact factor: 5.156
Authors: Christina Hertel; Dmytro Fishman; Anna Lorenc; Annamari Ranki; Kai Krohn; Pärt Peterson; Kai Kisand; Adrian Hayday Journal: Elife Date: 2019-06-27 Impact factor: 8.140
Authors: Alice Lepelley; Maria José Martin-Niclós; Melvin Le Bihan; Joseph A Marsh; Carolina Uggenti; Gillian I Rice; Vincent Bondet; Darragh Duffy; Jonny Hertzog; Jan Rehwinkel; Serge Amselem; Siham Boulisfane-El Khalifi; Mary Brennan; Edwin Carter; Lucienne Chatenoud; Stéphanie Chhun; Aurore Coulomb l'Hermine; Marine Depp; Marie Legendre; Karen J Mackenzie; Jonathan Marey; Catherine McDougall; Kathryn J McKenzie; Thierry Jo Molina; Bénédicte Neven; Luis Seabra; Caroline Thumerelle; Marie Wislez; Nadia Nathan; Nicolas Manel; Yanick J Crow; Marie-Louise Frémond Journal: J Exp Med Date: 2020-11-02 Impact factor: 14.307