Significance

This report reveals a mechanism by which cigarette smoke (CS) could exacerbate local inflammatory disease. CS is a key environmental pollutant affecting millions of people globally and continues to be of considerable interest to the biomedical communities. We found that CS activates the AhR on Th17 cells, leading to the up-regulation of miR-132, which is then packaged into extracellular vesicles that induce osteoclastogenesis via the suppression of Cox2 that catalyzes prostaglandins. Clinically, rheumatoid arthritis (RA) patients who smoke express a higher level of miRNA-132 compared to nonsmoking RA patients. This finding not only reveals a mechanism of CS signaling but also may provide a potential target for therapeutic intervention for inflammatory disease in general and RA in particular.

Abstract

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by joint destruction and severe morbidity. Cigarette smoking (CS) can exacerbate the incidence and severity of RA. Although Th17 cells and the Aryl hydrocarbon receptor (AhR) have been implicated, the mechanism by which CS induces RA development remains unclear. Here, using transcriptomic analysis, we show that microRNA-132 is specifically induced in Th17 cells in the presence of either AhR agonist or CS-enriched medium. miRNA-132 thus induced is packaged into extracellular vesicles produced by Th17 and acts as a proinflammatory mediator increasing osteoclastogenesis through the down-regulation of COX2. In vivo, articular knockdown of miR-132 in murine arthritis models reduces the number of osteoclasts in the joints. Clinically, RA patients express higher levels of miR-132 than do healthy individuals. This increase is further elevated by cigarette smoking. Together, these results reveal a hitherto unrecognized mechanism by which CS could exacerbate RA and further advance understanding of the impact of environmental factors on the pathogenesis of chronic inflammatory diseases.

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Data Availability

Microarray data analysis data have been deposited in the Gene Expression Omnibus database (GSE149169).

Acknowledgments

We thank Sergio L. C. Almeida, Patricia Rolim, and members of the Rheumatoid Arthritis team for their advice and clinical assistance; Dr. Fausto Almeida research group for support with Nanosight analysis; Ieda R. S. Schivo, Sergio R. Rosa, Ana Katia dos Santos, Denise Ferraz, and Tadeu F. Vieira (in memory) for technical assistance. This work was supported by grants from the São Paulo Research Foundation (FAPESP, 2012/02438-0 and 2013/08216-2 Center for Research in Inflammatory Disease) and from the University of São Paulo (NAP-DIN, 11.1.21625.01.0).

Supporting Information

Appendix (PDF)

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Information & Authors

Information

Published in

The cover image for PNAS Vol.118; No.1
Proceedings of the National Academy of Sciences
Vol. 118 | No. 1
January 5, 2021
PubMed: 33443169

Classifications

Data Availability

Microarray data analysis data have been deposited in the Gene Expression Omnibus database (GSE149169).

Submission history

Received: August 15, 2020
Accepted: November 18, 2020
Published online: December 28, 2020
Published in issue: January 5, 2021

Change history

October 18, 2021: Figure 3 and the SI Appendix have been updated; please see accompanying Correction for details. Previous version (December 28, 2020)

Keywords

  1. cigarette smoke
  2. rheumatoid arthritis
  3. Th17
  4. exosomes
  5. osteoclastogenesis

Acknowledgments

We thank Sergio L. C. Almeida, Patricia Rolim, and members of the Rheumatoid Arthritis team for their advice and clinical assistance; Dr. Fausto Almeida research group for support with Nanosight analysis; Ieda R. S. Schivo, Sergio R. Rosa, Ana Katia dos Santos, Denise Ferraz, and Tadeu F. Vieira (in memory) for technical assistance. This work was supported by grants from the São Paulo Research Foundation (FAPESP, 2012/02438-0 and 2013/08216-2 Center for Research in Inflammatory Disease) and from the University of São Paulo (NAP-DIN, 11.1.21625.01.0).

Notes

This article is a PNAS Direct Submission.

Authors

Affiliations

Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, CEP 14049-900, Brazil;
Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, CEP 14049-900, Brazil;
Raphael S. Peres
Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, CEP 14049-900, Brazil;
Fausto Almeida
Departament of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, CEP 14049-900, Brazil;
Sandra Y. Fukada
Department of BioMolecular Sciences, School of Pharmaceutical Sciences, University of São Paulo, São Paulo, SP, CEP 14040-903, Brazil;
Tarcilia A. Silva
Department of Oral Surgery and Pathology, School of Dentistry, Universidade Federal de Minas Gerais, Belo Horizonte, MG, CEP 31270-901, Brazil;
Daniele C. Nascimento
Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, CEP 14049-900, Brazil;
Nerry T. Cecilio
Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, CEP 14049-900, Brazil;
Jhimmy Talbot
Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, CEP 14049-900, Brazil;
Department of Internal Medicine, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, CEP 14049-900, Brazil;
Molecular Immunogenetics Group, Department of Genetics, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, CEP 14049-900, Brazil;
José Carlos Alves-Filho
Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, CEP 14049-900, Brazil;
Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, CEP 14049-900, Brazil;
Department of Internal Medicine, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, CEP 14049-900, Brazil;
Division of Immunology, Infection and Inflammation, Glasgow Biomedical Research Centre, University of Glasgow, Glasgow G12 8TA, United Kingdom
Fernando Q. Cunha1 [email protected]
Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, CEP 14049-900, Brazil;

Notes

1
To whom correspondence may be addressed. Email: [email protected] or [email protected].
Author contributions: P.B.D., S.Y.F., T.A.S., R.D.O., J.C.A.-F., T.M.C., P.L.-J., F.Y.L., and F.Q.C. designed research; F.Y.L. and F.Q.C. supervised the project; P.B.D., K.A.d.L., R.S.P., F.A., D.C.N., N.T.C., and J.T. performed research; G.A.P. contributed new reagents/analytic tools; P.B.D., K.A.d.L., R.S.P., F.A., S.Y.F., T.A.S., D.C.N., N.T.C., J.T., R.D.O., J.C.A.-F., T.M.C., P.L.-J., F.Y.L., and F.Q.C. analyzed data; and P.B.D., F.Y.L., and F.Q.C. wrote the paper.

Competing Interests

The authors declare no competing interest.

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    Cigarette smoke induces miR-132 in Th17 cells that enhance osteoclastogenesis in inflammatory arthritis
    Proceedings of the National Academy of Sciences
    • Vol. 118
    • No. 1

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