title: Twist1 and Twist2 Contribute to Cytokine Downregulation following Chronic NOD2 Stimulation of Human Macrophages through the Coordinated Regulation of Transcriptional Repressors and Activators.
abstract: Proper regulation of microbial-induced cytokines is critical to intestinal immune homeostasis. Acute stimulation of nucleotide-binding oligomerization domain 2 (NOD2), the Crohn's disease-associated sensor of bacterial peptidoglycan, induces cytokines. However, chronic NOD2 stimulation in macrophages decreases cytokines upon pattern recognition receptor (PRR) restimulation; cytokine attenuation to PRR stimulation is similarly observed in intestinal macrophages. The role for the transcriptional repressors Twist1 and Twist2 in regulating PRR-induced cytokine outcomes is poorly understood and has not been reported for NOD2. We found that Twist1 and Twist2 were required for optimal cytokine downregulation during acute and, particularly, chronic NOD2 stimulation of human macrophages. Consistently, Twist1 and Twist2 expression was increased after chronic NOD2 stimulation; this increased expression was IL-10 and TGF-β dependent. Although Twist1 and Twist2 did not coregulate each other's expression, they cooperated to enhance binding to cytokine promoters after chronic NOD2 stimulation. Moreover, Twist1 and Twist2 contributed to enhance expression and promoter binding of the proinflammatory inhibitor c-Maf and the transcriptional repressor Bmi1. Restoring c-Maf and Bmi1 expression in Twist-deficient macrophages restored NOD2-induced cytokine downregulation. Furthermore, with chronic NOD2 stimulation, Twist1 and Twist2 contributed to the decreased expression and cytokine promoter binding of the transcriptional activators activating transcription factor 4, C/EBPα, Runx1, and Runx2. Knockdown of these transcriptional activators in Twist-deficient macrophages restored cytokine downregulation after chronic NOD2 stimulation. Finally, NOD2 synergized with additional PRRs to increase Twist1 and Twist2 expression and Twist-dependent pathways. Therefore, after chronic NOD2 stimulation Twist1 and Twist2 coordinate the regulation of both transcriptional activators and repressors, thereby mediating optimal cytokine downregulation.
authors.affiliation: Department of Internal Medicine, Yale University, New Haven, CT 06520.
authors.forename: Shasha
authors.initials: S
authors.lastname: Zheng
authors.affiliation: Department of Internal Medicine, Yale University, New Haven, CT 06520.
authors.forename: Matija
authors.initials: M
authors.lastname: Hedl
authors.affiliation: Department of Internal Medicine, Yale University, New Haven, CT 06520 clara.abraham@yale.edu.
authors.forename: Clara
authors.initials: C
authors.lastname: Abraham
mesh_terms: D000119:Acetylmuramyl-Alanyl-Isoglutamine / Q000494:pharmacology; D051701:Activating Transcription Factor 4 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000494:pharmacology; D022762:CCAAT-Enhancer-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D050659:Core Binding Factor Alpha 1 Subunit / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D050676:Core Binding Factor Alpha 2 Subunit / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D005786:Gene Expression Regulation; D006801:Humans; D016753:Interleukin-10 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D008262:Macrophage Activation / Q000187:drug effects*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D053473:Nod2 Signaling Adaptor Protein / Q000819:agonists / Q000235:genetics / Q000276:immunology*; D009687:Nuclear Proteins / Q000235:genetics / Q000276:immunology*; D063150:Polycomb Repressive Complex 1 / Q000235:genetics / Q000276:immunology; D061251:Primary Cell Culture; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D051257:Proto-Oncogene Proteins c-maf / Q000235:genetics / Q000276:immunology; D034741:RNA, Small Interfering / Q000235:genetics / Q000276:immunology; D012097:Repressor Proteins / Q000235:genetics / Q000276:immunology*; D015398:Signal Transduction; D014158:Transcription, Genetic; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D051879:Twist-Related Protein 1 / Q000235:genetics / Q000276:immunology*
chemical_list: C497109:ATF4 protein, human; D057134:Antibodies, Neutralizing; C069077:BMI1 protein, human; D022762:CCAAT-Enhancer-Binding Proteins; C515848:CEBPA protein, human; D050659:Core Binding Factor Alpha 1 Subunit; D050676:Core Binding Factor Alpha 2 Subunit; C508609:IL10 protein, human; C495607:MAF protein, human; C429018:NOD2 protein, human; D053473:Nod2 Signaling Adaptor Protein; D009687:Nuclear Proteins; D051257:Proto-Oncogene Proteins c-maf; D034741:RNA, Small Interfering; C493728:RUNX1 protein, human; C493709:RUNX2 protein, human; D012097:Repressor Proteins; C103173:TWIST1 protein, human; C486274:TWIST2 protein, human; D016212:Transforming Growth Factor beta; D051879:Twist-Related Protein 1; D016753:Interleukin-10; D051701:Activating Transcription Factor 4; D000119:Acetylmuramyl-Alanyl-Isoglutamine; D063150:Polycomb Repressive Complex 1