A genome-wide siRNA screen reveals positive and negative regulators of the NOD2 and NF-κB signaling pathways.

Abstract

The cytoplasmic receptor NOD2 (nucleotide-binding oligomerization domain 2) senses peptidoglycan fragments and triggers host defense pathways, including activation of nuclear factor κB (NF-κB) signaling, which lead to inflammatory immune responses. Dysregulation of NOD2 signaling is associated with inflammatory diseases, such as Crohn's disease and Blau syndrome. We used a genome-wide small interfering RNA screen to identify regulators of the NOD2 signaling pathway. Several genes associated with Crohn's disease risk were identified in the screen. A comparison of candidates from this screen with other "omics" data sets revealed interconnected networks of genes implicated in NF-κB signaling, thus supporting a role for NOD2 and NF-κB pathways in the pathogenesis of Crohn's disease. Many of these regulators were validated in secondary assays, such as measurement of interleukin-8 secretion, which is partially dependent on NF-κB. Knockdown of putative regulators in human embryonic kidney 293 cells followed by stimulation with tumor necrosis factor-α revealed that most of the genes identified were general regulators of NF-κB signaling. Overall, the genes identified here provide a resource to facilitate the elucidation of the molecular mechanisms that regulate NOD2- and NF-κB-mediated inflammation.

• Authors: Neil Warner, Aaron Burberry, Luigi Franchi, Yun-Gi Kim, Christine McDonald, Maureen A Sartor, Gabriel Núñez
• Journal: Science signaling (Sci Signal) Vol. 6 Issue 258 Pg. rs3 (Jan 15 2013) ISSN: 1937-9145 [Electronic] United States
• PMID: 23322906 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• I-kappa B Proteins
• NF-kappa B
• NOD2 protein, human
• Nod2 Signaling Adaptor Protein
• RNA, Small Interfering
• Tumor Necrosis Factor-alpha
• Acetylmuramyl-Alanyl-Isoglutamine
• NADPH Oxidases
• neutrophil cytosolic factor 1
• Mitogen-Activated Protein Kinases

Topics

• Acetylmuramyl-Alanyl-Isoglutamine (pharmacology)
• Animals
• Blotting, Western
• Bone Marrow Cells (cytology, drug effects, metabolism)
• Cell Survival (genetics)
• Cells, Cultured
• Crohn Disease (genetics)
• Gene Expression Regulation (drug effects)
• Gene Regulatory Networks (drug effects)
• Genome, Human (genetics)
• HEK293 Cells
• Humans
• I-kappa B Proteins (metabolism)
• Macrophages (cytology, drug effects, metabolism)
• Mice
• Mice, Knockout
• Mitogen-Activated Protein Kinases (metabolism)
• Models, Genetic
• NADPH Oxidases (deficiency, genetics)
• NF-kappa B (genetics, metabolism)
• Nod2 Signaling Adaptor Protein (genetics, metabolism)
• RNA Interference
• RNA, Small Interfering (genetics)
• Signal Transduction (drug effects, genetics)
• Tumor Necrosis Factor-alpha (pharmacology)