Attenuating ABHD17 Isoforms Augments the S-acylation and Function of NOD2 and a Subset of Crohn’s Disease-associated NOD2 VariantsSummary

Background & Aims: NOD2 is an intracellular innate immune receptor that detects bacterial peptidoglycan fragments. Although nominally soluble, some NOD2 is associated with the plasma membrane and endosomal compartments for microbial surveillance. This membrane targeting is achieved through p...

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Main Authors: Charneal L. Dixon, Noah R. Martin, Micah J. Niphakis, Benjamin F. Cravatt, Gregory D. Fairn
Format: Article
Language:English
Published: Elsevier 2025-01-01
Series:Cellular and Molecular Gastroenterology and Hepatology
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Online Access:http://www.sciencedirect.com/science/article/pii/S2352345X25000323
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author Charneal L. Dixon
Noah R. Martin
Micah J. Niphakis
Benjamin F. Cravatt
Gregory D. Fairn
author_facet Charneal L. Dixon
Noah R. Martin
Micah J. Niphakis
Benjamin F. Cravatt
Gregory D. Fairn
author_sort Charneal L. Dixon
collection DOAJ
description Background & Aims: NOD2 is an intracellular innate immune receptor that detects bacterial peptidoglycan fragments. Although nominally soluble, some NOD2 is associated with the plasma membrane and endosomal compartments for microbial surveillance. This membrane targeting is achieved through post-translational S-acylation of NOD2 by the protein acyltransferase ZDHHC5. Membrane attachment is necessary to initiate a signaling cascade in response to cytosolic peptidoglycan fragments. Ultimately, this signaling results in the production of antimicrobial peptides and proinflammatory cytokines. In most cases, S-acylation is a reversible post-translational modification with removal of the fatty acyl chain catalyzed by one of several acyl protein thioesterases. Deacylation of NOD2 by such an enzyme will displace it from the plasma membrane and endosomes, thus preventing signaling. Methods: To identify the enzymes responsible for NOD2 deacylation, we used engineered cell lines with RNA interference and small-molecule inhibitors. These approaches were combined with confocal microscopy, acyl-resin-assisted capture, immunoblotting, and cytokine multiplex assays. Results: We identified α/β-hydrolase domain-containing protein 17 isoforms (ABHD17A, ABHD17B, and ABHD17C) as the acyl protein thioesterases responsible for NOD2 deacylation. Inhibiting ABHD17 increased the plasma membrane localization of wild-type NOD2 and a subset of poorly acylated Crohn’s disease-associated variants. This enhanced NOD2 activity, increasing NF-κB activation and pro-inflammatory cytokine production in epithelial cells. Conclusions: These findings demonstrate that ABHD17 isoforms are negative regulators of NOD2. The results also suggest that targeting ABHD17 isoforms could restore functionality to specific Crohn’s disease-associated NOD2 variants, offering a potential therapeutic strategy.
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spelling doaj-art-a7a24924b45d4e499e3e0db3137b7fdd2025-08-20T03:08:42ZengElsevierCellular and Molecular Gastroenterology and Hepatology2352-345X2025-01-0119610149110.1016/j.jcmgh.2025.101491Attenuating ABHD17 Isoforms Augments the S-acylation and Function of NOD2 and a Subset of Crohn’s Disease-associated NOD2 VariantsSummaryCharneal L. Dixon0Noah R. Martin1Micah J. Niphakis2Benjamin F. Cravatt3Gregory D. Fairn4Department of Pathology, Dalhousie University, Halifax, Nova Scotia, Canada; Keenan Research Centre for Biomedical Science, St Michael’s Hospital, Unity Health Toronto, Toronto, Ontario, CanadaDepartment of Pathology, Dalhousie University, Halifax, Nova Scotia, CanadaLundbeck La Jolla Research Center, Inc, San Diego, CaliforniaDepartment of Chemistry, The Scripps Research Institute, La Jolla, CaliforniaDepartment of Pathology, Dalhousie University, Halifax, Nova Scotia, Canada; Keenan Research Centre for Biomedical Science, St Michael’s Hospital, Unity Health Toronto, Toronto, Ontario, Canada; Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, Nova Scotia, Canada; Correspondence Address correspondence to: Gregory D. Fairn, Department of Pathology, Dalhousie University, Sir Charles Tupper Medical Building, Room 11B, 5850 College Street, PO Box 15000, Halifax, Nova Scotia, Canada B3H 4R2; tel: (902) 221-3719.Background & Aims: NOD2 is an intracellular innate immune receptor that detects bacterial peptidoglycan fragments. Although nominally soluble, some NOD2 is associated with the plasma membrane and endosomal compartments for microbial surveillance. This membrane targeting is achieved through post-translational S-acylation of NOD2 by the protein acyltransferase ZDHHC5. Membrane attachment is necessary to initiate a signaling cascade in response to cytosolic peptidoglycan fragments. Ultimately, this signaling results in the production of antimicrobial peptides and proinflammatory cytokines. In most cases, S-acylation is a reversible post-translational modification with removal of the fatty acyl chain catalyzed by one of several acyl protein thioesterases. Deacylation of NOD2 by such an enzyme will displace it from the plasma membrane and endosomes, thus preventing signaling. Methods: To identify the enzymes responsible for NOD2 deacylation, we used engineered cell lines with RNA interference and small-molecule inhibitors. These approaches were combined with confocal microscopy, acyl-resin-assisted capture, immunoblotting, and cytokine multiplex assays. Results: We identified α/β-hydrolase domain-containing protein 17 isoforms (ABHD17A, ABHD17B, and ABHD17C) as the acyl protein thioesterases responsible for NOD2 deacylation. Inhibiting ABHD17 increased the plasma membrane localization of wild-type NOD2 and a subset of poorly acylated Crohn’s disease-associated variants. This enhanced NOD2 activity, increasing NF-κB activation and pro-inflammatory cytokine production in epithelial cells. Conclusions: These findings demonstrate that ABHD17 isoforms are negative regulators of NOD2. The results also suggest that targeting ABHD17 isoforms could restore functionality to specific Crohn’s disease-associated NOD2 variants, offering a potential therapeutic strategy.http://www.sciencedirect.com/science/article/pii/S2352345X25000323ABHD17Acyl Protein ThioesteraseCrohn’s DiseaseIL-8InflammationNOD2
spellingShingle Charneal L. Dixon
Noah R. Martin
Micah J. Niphakis
Benjamin F. Cravatt
Gregory D. Fairn
Attenuating ABHD17 Isoforms Augments the S-acylation and Function of NOD2 and a Subset of Crohn’s Disease-associated NOD2 VariantsSummary
Cellular and Molecular Gastroenterology and Hepatology
ABHD17
Acyl Protein Thioesterase
Crohn’s Disease
IL-8
Inflammation
NOD2
title Attenuating ABHD17 Isoforms Augments the S-acylation and Function of NOD2 and a Subset of Crohn’s Disease-associated NOD2 VariantsSummary
title_full Attenuating ABHD17 Isoforms Augments the S-acylation and Function of NOD2 and a Subset of Crohn’s Disease-associated NOD2 VariantsSummary
title_fullStr Attenuating ABHD17 Isoforms Augments the S-acylation and Function of NOD2 and a Subset of Crohn’s Disease-associated NOD2 VariantsSummary
title_full_unstemmed Attenuating ABHD17 Isoforms Augments the S-acylation and Function of NOD2 and a Subset of Crohn’s Disease-associated NOD2 VariantsSummary
title_short Attenuating ABHD17 Isoforms Augments the S-acylation and Function of NOD2 and a Subset of Crohn’s Disease-associated NOD2 VariantsSummary
title_sort attenuating abhd17 isoforms augments the s acylation and function of nod2 and a subset of crohn s disease associated nod2 variantssummary
topic ABHD17
Acyl Protein Thioesterase
Crohn’s Disease
IL-8
Inflammation
NOD2
url http://www.sciencedirect.com/science/article/pii/S2352345X25000323
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