Impaired microRNA processing in neutrophils from rheumatoid arthritis patients confers their pathogenic profile. Modulation by biological therapies

Haematologica. 2020 Sep 1;105(9):2250-2261. doi: 10.3324/haematol.2018.205047.

Abstract

The aim of this study was to investigate the microRNA (miRNA) expression pattern in neutrophils from rheumatoid arthritis (RA) patients and its contribution to their pathogenic profile and to analyze the effect of specific autoantibodies or inflammatory components in the regulation of miRNA in RA neutrophils and its modulation by biological therapies. Neutrophils were isolated from paired peripheral blood (PB) and synovial fluid samples of 40 patients with RA and from PB of 40 healthy donors. A miRNA array was performed using nCounter technology. Neutrophils from healthy donors were treated in vitrowith antibodies to citrullinated protein antigens isolated from RA patients and tumor necrosis factor-a (TNF-a) or interleukin-6. A number of cytokines and chemokines were analyzed. In vitro treatments of RA-neutrophils with tocilizumab or infliximab were carried out. Transfections with pre-miRNA and DICER downregulation experiments were further performed. RA-neutrophils showed a global downregulation of miRNA and genes involved in their biogenesis, alongside with an upregulation of various potential mRNA targets related to migration and inflammation. Decreased levels of miRNA and DICER correlated with autoimmunity, inflammation and disease activity. Citrullinated protein antigens and TNF-a decreased the expression of numerous miRNA and their biogenesis-related genes, increasing their potential mRNA targets. Infliximab reversed those effects. Transfections with pre-miRNA-223, -126 and -148a specifically modulated genes regulating inflammation, survival and migration whereas DICER depletion influenced the inflammatory profile of neutrophils. Taken together RA-neutrophils exhibited a global low abundance of miRNA induced by autoantibodies and inflammatory markers, which potentially contributed to their pathogenic activation. miRNA biogenesis was significantly impaired in RAneutrophils and further associated with a greater downregulation of miRNA mainly related to migration and inflammation in synovial fluid neutrophils. Finally, anti-TNF-a and anti-interleukin-6 receptor treatments can modulate miRNA levels in the neutrophils, minimizing their inflammatory profile.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arthritis, Rheumatoid* / drug therapy
  • Arthritis, Rheumatoid* / genetics
  • Biological Therapy
  • Humans
  • MicroRNAs* / genetics
  • Neutrophils
  • Tumor Necrosis Factor-alpha / genetics

Substances

  • MicroRNAs
  • Tumor Necrosis Factor-alpha

Grants and funding

Funding: This work was supported by grants from the Instituto de Salud Carlos III (CP15/00158, PI17/01316 and PI18/00837), cofinanciado por el fondo europeo de desarrollo regional de la Union Europea, ‘una manera de hacer Europa’, Spain, the Regional Health System (ref. PI 0285 2017), and the Spanish Inflammatory and Rheumatic Diseases Network (RIER, RD16/0012/0015). CL-P was supported by contracts from both the Junta de Andalucia and Spanish Ministry of Science and Innovation (Ramon y Cajal). NB was supported by Ministry of Health postdoctoral fellowship (Miguel Servet Programme). YJ-G was supported by a contract from the University of Cordoba [co-funded by the Research Plan of the University of Cordoba and the Operating Program of the European Regional Development Funds (ERDF) for Andalusia. CL-P was supported by a contract from the Junta de Andalucia] (Nicolas Monardes Programme). YJ-G was supported by a contract from the University of Cordoba (co-funded by the Research Plan of the University of Cordoba and the Operating Program of the European Regional Development Funds [ERDF]) for Andalusia).