Ana Rebane1, Toomas Runnel2, Alar Aab3, Julia Maslovskaja3, Beate Rückert4, Maya Zimmermann4, Mario Plaas5, Jaanika Kärner3, Angela Treis4, Maire Pihlap6, Uku Haljasorg6, Helen Hermann6, Nikoletta Nagy7, Lajos Kemeny7, Triin Erm8, Külli Kingo9, Mei Li10, Mark P Boldin11, Cezmi A Akdis4. 1. Swiss Institute of Allergy and Asthma Research (SIAF), University of Zürich, Davos, Switzerland; Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia. Electronic address: ana.rebane@ut.ee. 2. Swiss Institute of Allergy and Asthma Research (SIAF), University of Zürich, Davos, Switzerland; Institute of Molecular and Cellular Biology, University of Tartu, Tartu, Estonia. 3. Swiss Institute of Allergy and Asthma Research (SIAF), University of Zürich, Davos, Switzerland; Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia. 4. Swiss Institute of Allergy and Asthma Research (SIAF), University of Zürich, Davos, Switzerland. 5. Transgenic Technology Core Laboratory, University of Tartu, Tartu, Estonia. 6. Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia. 7. Department of Dermatology and Allergology, University of Szeged, Szeged, Hungary; Dermatological Research Group of the Hungarian Academy of Sciences, Szeged, Hungary. 8. Department of Pathology, Tartu University Hospital, Tartu, Estonia. 9. Department of Dermatology and Venereology, University of Tartu, Tartu, Estonia; Dermatology Clinic, Tartu University Hospital, Tartu, Estonia. 10. Institut de Génétique et de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique/Institut National de la Santé et de la Recherche Médicale/Université de Strasbourg, Illkirch, France. 11. Department of Molecular and Cellular Biology, Beckman Research Institute, City of Hope, Duarte, Calif.
Abstract
BACKGROUND: Chronic skin inflammation in atopic dermatitis (AD) is associated with elevated expression of proinflammatory genes and activation of innate immune responses in keratinocytes. microRNAs (miRNAs) are short, single-stranded RNA molecules that silence genes via the degradation of target mRNAs or inhibition of translation. OBJECTIVE: The aim of this study was to investigate the role of miR-146a in skin inflammation in AD. METHODS: RNA and protein expression was analyzed using miRNA and mRNA arrays, RT-quantitative PCR, Western blotting, and immunonohistochemistry. Transfection of miR-146a precursors and inhibitors into human primary keratinocytes, luciferase assays, and MC903-dependent mouse model of AD were used to study miR-146a function. RESULTS: We show that miR-146a expression is increased in keratinocytes and chronic lesional skin of patients with AD. miR-146a inhibited the expression of numerous proinflammatory factors, including IFN-γ-inducible and AD-associated genes CCL5, CCL8, and ubiquitin D (UBD) in human primary keratinocytes stimulated with IFN-γ, TNF-α, or IL-1β. In a mouse model of AD, miR-146a-deficient mice developed stronger inflammation characterized by increased accumulation of infiltrating cells in the dermis, elevated expression of IFN-γ, CCL5, CCL8, and UBD in the skin, and IFN-γ, IL-1β, and UBD in draining lymph nodes. Both tissue culture and in vivo experiments in mice demonstrated that miR-146a-mediated suppression in allergic skin inflammation partially occurs through direct targeting of upstream nuclear factor kappa B signal transducers caspase recruitment domain-containing protein 10 and IL-1 receptor-associated kinase 1. In addition, human CCL5 was determined as a novel, direct target of miR-146a. CONCLUSION: Our data demonstrate that miR-146a controls nuclear factor kappa B-dependent inflammatory responses in keratinocytes and chronic skin inflammation in AD.
BACKGROUND:Chronic skin inflammation in atopic dermatitis (AD) is associated with elevated expression of proinflammatory genes and activation of innate immune responses in keratinocytes. microRNAs (miRNAs) are short, single-stranded RNA molecules that silence genes via the degradation of target mRNAs or inhibition of translation. OBJECTIVE: The aim of this study was to investigate the role of miR-146a in skin inflammation in AD. METHODS: RNA and protein expression was analyzed using miRNA and mRNA arrays, RT-quantitative PCR, Western blotting, and immunonohistochemistry. Transfection of miR-146a precursors and inhibitors into human primary keratinocytes, luciferase assays, and MC903-dependent mouse model of AD were used to study miR-146a function. RESULTS: We show that miR-146a expression is increased in keratinocytes and chronic lesional skin of patients with AD. miR-146a inhibited the expression of numerous proinflammatory factors, including IFN-γ-inducible and AD-associated genes CCL5, CCL8, and ubiquitin D (UBD) in human primary keratinocytes stimulated with IFN-γ, TNF-α, or IL-1β. In a mouse model of AD, miR-146a-deficientmice developed stronger inflammation characterized by increased accumulation of infiltrating cells in the dermis, elevated expression of IFN-γ, CCL5, CCL8, and UBD in the skin, and IFN-γ, IL-1β, and UBD in draining lymph nodes. Both tissue culture and in vivo experiments in mice demonstrated that miR-146a-mediated suppression in allergic skin inflammation partially occurs through direct targeting of upstream nuclear factor kappa B signal transducers caspase recruitment domain-containing protein 10 and IL-1 receptor-associated kinase 1. In addition, humanCCL5 was determined as a novel, direct target of miR-146a. CONCLUSION: Our data demonstrate that miR-146a controls nuclear factor kappa B-dependent inflammatory responses in keratinocytes and chronic skin inflammation in AD.
Authors: Helen Vaher; Toomas Runnel; Egon Urgard; Alar Aab; Gemma Carreras Badosa; Julia Maslovskaja; Kristi Abram; Liisi Raam; Bret Kaldvee; Tarmo Annilo; Eric R Tkaczyk; Toivo Maimets; Cezmi A Akdis; Külli Kingo; Ana Rebane Journal: Allergy Date: 2019-06-06 Impact factor: 13.146
Authors: Helen Hermann; Toomas Runnel; Alar Aab; Hansjörg Baurecht; Elke Rodriguez; Nathaniel Magilnick; Egon Urgard; Liisi Šahmatova; Ele Prans; Julia Maslovskaja; Kristi Abram; Maire Karelson; Bret Kaldvee; Paula Reemann; Uku Haljasorg; Beate Rückert; Paulina Wawrzyniak; Michael Weichenthal; Ulrich Mrowietz; Andre Franke; Christian Gieger; Jonathan Barker; Richard Trembath; Lam C Tsoi; James T Elder; Eric R Tkaczyk; Kai Kisand; Pärt Peterson; Külli Kingo; Mark Boldin; Stephan Weidinger; Cezmi A Akdis; Ana Rebane Journal: J Invest Dermatol Date: 2017-06-06 Impact factor: 8.551
Authors: Ian A Myles; Kelli W Williams; Jensen D Reckhow; Momodou L Jammeh; Nathan B Pincus; Inka Sastalla; Danial Saleem; Kelly D Stone; Sandip K Datta Journal: JCI Insight Date: 2016-07-07