An off-target effect of class A CpG-oligonucleotides on suppressing the cyclic GMP-AMP synthase signaling in fibroblastic reticular cells

BackgroundClass A CpG-oligonucleotides (ODNs), a Toll-like receptor 9 (TLR9) agonist, have been applied for treating inflammatory diseases and cancer in preclinical studies and clinical trials. A recent study has reported that class A ODNs can activate the Cyclic GMP-AMP synthase (cGAS) signaling to...

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Main Authors: Preethi Jayakumar, Ting Jiang, Hai Huang, Meihong Deng
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-04-01
Series:Frontiers in Pharmacology
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Online Access:https://www.frontiersin.org/articles/10.3389/fphar.2025.1576151/full
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author Preethi Jayakumar
Preethi Jayakumar
Ting Jiang
Hai Huang
Hai Huang
Hai Huang
Meihong Deng
Meihong Deng
Meihong Deng
author_facet Preethi Jayakumar
Preethi Jayakumar
Ting Jiang
Hai Huang
Hai Huang
Hai Huang
Meihong Deng
Meihong Deng
Meihong Deng
author_sort Preethi Jayakumar
collection DOAJ
description BackgroundClass A CpG-oligonucleotides (ODNs), a Toll-like receptor 9 (TLR9) agonist, have been applied for treating inflammatory diseases and cancer in preclinical studies and clinical trials. A recent study has reported that class A ODNs can activate the Cyclic GMP-AMP synthase (cGAS) signaling to regulate the inflammatory response in human monocytes. However, it remains unknown whether class A ODNs can activate the cGAS pathways in other cell types, such as fibroblastic reticular cells (FRC), which play critical roles in modulating the immune environments during inflammatory diseases and cancer.MethodsTo understand the role of class A ODN in regulating the cGAS signaling in FRC, we treated mouse FRC and human fibroblast with class A ODN, a cGAS agonist (HT-DNA), and combined class A and HT-DNA.ResultsUnexpectedly, we found that class A ODNs suppress the cGAS level and downstream signaling in human and murine FRC. The class A ODN-induced suppression effect on cGAS is limited in FRC, but not other immune cell types, and is independent of TLR9. Performing pulldown assay and Mass spectrum, we found that class A ODNs regulate the cGAS level post translationally by interacting with cGAS and ZNF598, an E3 ubiquitin ligase.ConclusionOur data reveal an unrecognized off-target effect of class A ODN on suppressing the cGAS signaling in FRCs, which should be considered when designing class A ODN regimens for inflammatory diseases and cancer.
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spelling doaj-art-77c8fe6db9b74b619f69b6c4cc7deb7a2025-08-20T03:13:21ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122025-04-011610.3389/fphar.2025.15761511576151An off-target effect of class A CpG-oligonucleotides on suppressing the cyclic GMP-AMP synthase signaling in fibroblastic reticular cellsPreethi Jayakumar0Preethi Jayakumar1Ting Jiang2Hai Huang3Hai Huang4Hai Huang5Meihong Deng6Meihong Deng7Meihong Deng8Center for Immunology and Inflammation, The Feinstein Institutes for Medical Research, Manhasset, NY, United StatesInstitute of Molecular Medicine, Feinstein Institutes for Medical Research, New York, NY, United StatesTsinghua University School of Medicine, Beijing, ChinaCenter for Immunology and Inflammation, The Feinstein Institutes for Medical Research, Manhasset, NY, United StatesInstitute of Molecular Medicine, Feinstein Institutes for Medical Research, New York, NY, United StatesDepartments of Molecular Medicine and Surgery, Zucker School of Medicine at Hofstra University/Northwell, Hempstead, NY, United StatesCenter for Immunology and Inflammation, The Feinstein Institutes for Medical Research, Manhasset, NY, United StatesInstitute of Molecular Medicine, Feinstein Institutes for Medical Research, New York, NY, United StatesDepartments of Molecular Medicine and Surgery, Zucker School of Medicine at Hofstra University/Northwell, Hempstead, NY, United StatesBackgroundClass A CpG-oligonucleotides (ODNs), a Toll-like receptor 9 (TLR9) agonist, have been applied for treating inflammatory diseases and cancer in preclinical studies and clinical trials. A recent study has reported that class A ODNs can activate the Cyclic GMP-AMP synthase (cGAS) signaling to regulate the inflammatory response in human monocytes. However, it remains unknown whether class A ODNs can activate the cGAS pathways in other cell types, such as fibroblastic reticular cells (FRC), which play critical roles in modulating the immune environments during inflammatory diseases and cancer.MethodsTo understand the role of class A ODN in regulating the cGAS signaling in FRC, we treated mouse FRC and human fibroblast with class A ODN, a cGAS agonist (HT-DNA), and combined class A and HT-DNA.ResultsUnexpectedly, we found that class A ODNs suppress the cGAS level and downstream signaling in human and murine FRC. The class A ODN-induced suppression effect on cGAS is limited in FRC, but not other immune cell types, and is independent of TLR9. Performing pulldown assay and Mass spectrum, we found that class A ODNs regulate the cGAS level post translationally by interacting with cGAS and ZNF598, an E3 ubiquitin ligase.ConclusionOur data reveal an unrecognized off-target effect of class A ODN on suppressing the cGAS signaling in FRCs, which should be considered when designing class A ODN regimens for inflammatory diseases and cancer.https://www.frontiersin.org/articles/10.3389/fphar.2025.1576151/fullclass A ODNTLR9 agonistCGASZNF598fibroblastic reticular cells
spellingShingle Preethi Jayakumar
Preethi Jayakumar
Ting Jiang
Hai Huang
Hai Huang
Hai Huang
Meihong Deng
Meihong Deng
Meihong Deng
An off-target effect of class A CpG-oligonucleotides on suppressing the cyclic GMP-AMP synthase signaling in fibroblastic reticular cells
Frontiers in Pharmacology
class A ODN
TLR9 agonist
CGAS
ZNF598
fibroblastic reticular cells
title An off-target effect of class A CpG-oligonucleotides on suppressing the cyclic GMP-AMP synthase signaling in fibroblastic reticular cells
title_full An off-target effect of class A CpG-oligonucleotides on suppressing the cyclic GMP-AMP synthase signaling in fibroblastic reticular cells
title_fullStr An off-target effect of class A CpG-oligonucleotides on suppressing the cyclic GMP-AMP synthase signaling in fibroblastic reticular cells
title_full_unstemmed An off-target effect of class A CpG-oligonucleotides on suppressing the cyclic GMP-AMP synthase signaling in fibroblastic reticular cells
title_short An off-target effect of class A CpG-oligonucleotides on suppressing the cyclic GMP-AMP synthase signaling in fibroblastic reticular cells
title_sort off target effect of class a cpg oligonucleotides on suppressing the cyclic gmp amp synthase signaling in fibroblastic reticular cells
topic class A ODN
TLR9 agonist
CGAS
ZNF598
fibroblastic reticular cells
url https://www.frontiersin.org/articles/10.3389/fphar.2025.1576151/full
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