Cell-Free DNA Derived From Neutrophils Triggers Type 1 Interferon Signature in Neuromyelitis Optica Spectrum Disorder.


Journal

Neurology(R) neuroimmunology & neuroinflammation
ISSN: 2332-7812
Titre abrégé: Neurol Neuroimmunol Neuroinflamm
Pays: United States
ID NLM: 101636388

Informations de publication

Date de publication:
05 2022
Historique:
received: 27 10 2021
accepted: 12 01 2022
entrez: 25 2 2022
pubmed: 26 2 2022
medline: 28 4 2022
Statut: epublish

Résumé

Recently accumulating evidence suggests the pivotal role of type 1 interferon (IFN-1) signature in the pathogenesis of neuromyelitis optica spectrum disorder (NMOSD). However, the mechanism of the initial trigger that augments IFN-1 pathway in the peripheral immune system of NMOSD has yet to be elucidated. Clinical samples were obtained from 32 patients with aquaporin-4 antibody-positive NMOSD and 23 healthy subjects. IFN-1 induction in peripheral blood mononuclear cells (PBMCs) by serum-derived cell-free DNA (cfDNA) was assessed in combination with blockades of DNA sensors in vitro. CfDNA fraction was analyzed for DNA methylation profiles by bisulfite sequencing, elucidating the cellular origin of cfDNA. The induction of neutrophil extracellular trap related cell death (NETosis) was further analyzed in NMOSD and control groups, and the efficacy of pharmacologic intervention of NETosis was assessed. Enhanced IFN-1 induction by cfDNA derived from NMOSD was observed in PBMCs with cofactor of LL37 antimicrobial peptide. DNase treatment, cGAS inhibitor, and Toll-like receptor 9 antagonist efficiently inhibited IFN-1 production. DNA methylation pattern of cfDNA in patients with NMOSD demonstrated that the predominant cellular source of cfDNA was neutrophils. Whole blood transcriptome analysis also revealed neutrophil activation in NMOSD. In addition, enhanced NETosis induction was observed with NMOSD-derived sera, and efficient pharmacologic inhibition of NETosis with dipyridamole was observed. Our study highlights the previously unrevealed role of cfDNA predominantly released by neutrophil in the induction of IFN-1 signature in NMOSD and further indicate a novel pharmacologic target in NMOSD.

Sections du résumé

BACKGROUND AND OBJECTIVES
Recently accumulating evidence suggests the pivotal role of type 1 interferon (IFN-1) signature in the pathogenesis of neuromyelitis optica spectrum disorder (NMOSD). However, the mechanism of the initial trigger that augments IFN-1 pathway in the peripheral immune system of NMOSD has yet to be elucidated.
METHODS
Clinical samples were obtained from 32 patients with aquaporin-4 antibody-positive NMOSD and 23 healthy subjects. IFN-1 induction in peripheral blood mononuclear cells (PBMCs) by serum-derived cell-free DNA (cfDNA) was assessed in combination with blockades of DNA sensors in vitro. CfDNA fraction was analyzed for DNA methylation profiles by bisulfite sequencing, elucidating the cellular origin of cfDNA. The induction of neutrophil extracellular trap related cell death (NETosis) was further analyzed in NMOSD and control groups, and the efficacy of pharmacologic intervention of NETosis was assessed.
RESULTS
Enhanced IFN-1 induction by cfDNA derived from NMOSD was observed in PBMCs with cofactor of LL37 antimicrobial peptide. DNase treatment, cGAS inhibitor, and Toll-like receptor 9 antagonist efficiently inhibited IFN-1 production. DNA methylation pattern of cfDNA in patients with NMOSD demonstrated that the predominant cellular source of cfDNA was neutrophils. Whole blood transcriptome analysis also revealed neutrophil activation in NMOSD. In addition, enhanced NETosis induction was observed with NMOSD-derived sera, and efficient pharmacologic inhibition of NETosis with dipyridamole was observed.
DISCUSSION
Our study highlights the previously unrevealed role of cfDNA predominantly released by neutrophil in the induction of IFN-1 signature in NMOSD and further indicate a novel pharmacologic target in NMOSD.

Identifiants

pubmed: 35210295
pii: 9/3/e1149
doi: 10.1212/NXI.0000000000001149
pmc: PMC8874356
pii:
doi:

Substances chimiques

Cell-Free Nucleic Acids 0
Interferons 9008-11-1

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Informations de copyright

Copyright © 2022 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the American Academy of Neurology.

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Auteurs

Hisashi Murata (H)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

Makoto Kinoshita (M)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan. mkinoshita@neurol.med.osaka-u.ac.jp.

Yoshiaki Yasumizu (Y)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

Daisuke Motooka (D)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

Shohei Beppu (S)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

Naoyuki Shiraishi (N)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

Yasuko Sugiyama (Y)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

Keigo Kihara (K)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

Satoru Tada (S)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

Toru Koda (T)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

Hachiro Konaka (H)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

Hyota Takamatsu (H)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

Atsushi Kumanogoh (A)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

Tatsusada Okuno (T)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

Hideki Mochizuki (H)

From the Department of Neurology (Hisashi Murata, M.K., Y.Y., S.B., N.S., Y.S., K.K., S.T., T.K., T.O., Hideki Mochizuki), Graduate School of Medicine, Osaka University; Department of Experimental Immunology (Y.Y.), WPI Immunology Frontier Research Center, Osaka University; Integrated Frontier Research for Medical Science Division (Y.Y., D.M., A.K., Hideki Mochizuki), Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University; Genome Information Research Center (D.M.), Research Institute for Microbial Diseases, Osaka University; and Department of Respiratory Medicine and Clinical Immunology (H.K., H.T., A.K.), Graduate School of Medicine, Osaka University, Suita, Japan.

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