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Mitochondrial mutagenesis correlates with the local inflammatory environment in arthritis
  1. Leonard C Harty1,
  2. Monika Biniecka1,
  3. Jacintha O'Sullivan2,
  4. Edward Fox3,
  5. Kevin Mulhall4,
  6. Douglas J Veale1,
  7. Ursula Fearon1
  1. 1Translation Rheumatology Research Group, Dublin Academic Medical Centre, St Vincent's University Hospital, Elm Park, Dublin, Ireland
  2. 2Department of Surgery, Institute of Molecular Medicine, Trinity College Dublin, St James's Hospital, Dublin, Ireland
  3. 3Department of Pathology, University of Washington, Seattle, Washington, USA
  4. 4Department of Orthopaedics, Dublin Academic Medical Centre, Mater Misericordiae University Hospital, Dublin, Ireland
  1. Correspondence to Dr Ursula Fearon, Translational Rheumatology Group, Dublin Academic Medical Centre, St Vincent's University Hospital and The Conway Institute of Biomolecular and Biomedical Research, Dublin 4, Ireland; ursula.fearon{at}ucd.ie

Abstract

Background To examine the association between mitochondrial mutagenesis and the proinflammatory microenvironment in patients with inflammatory arthritis.

Methods Fifty patients with inflammatory arthritis underwent arthroscopy and synovial tissue biopsies, synovial fluid and clinical assessment were obtained. Fifteen patients pre/post-TNFi therapy were also recruited. Normal synovial biopsies were obtained from 10 subjects undergoing interventional arthroscopy. Macroscopic synovitis/vascularity was measured by visual analogue scale. Cell-specific markers CD3 (T cells) and CD68 (macrophages) were quantified by immunohistology. TNFα, IL-6, IFNγ and IL-1β were measured in synovial fluids by MSD multiplex assays. Synovial tissue mitochondrial mutagenesis was quantified using a mitochondrial random mutation capture assay (RMCA). The direct effect of TNFα on oxidative stress and mitochondrial function was assessed in primary cultures of rheumatoid arthritis synovial fibroblast cells (RASFCs). Mitochondrial mutagenesis, reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and mitochondrial mass (MM) were quantified using the RMCA and specific cell fluorescent probes.

Results A significant increase in mtDNA mutation frequency was demonstrated in inflamed synovial tissue compared with control (p<0.05), an effect that was independent of age. mtDNA mutations positively correlated with macroscopic synovitis (r=0.52, p<0.016), vascularity (r=0.54, p<0.01) and with synovial fluid cytokine levels of TNFα (r=0.74, p<0.024) and IFNγ (r=0.72, p<0.039). mtDNA mutation frequency post-TNFi therapy was significantly lower in patients with a DAS<3.2 (p<0.05) and associated with clinical and microscopic measures of disease (p<0.05). In vitro TNFα significantly induced mtDNA mutations, ROS, MM and MMP in RASFCs (all p<0.05).

Conclusion High mitochondrial mutations are strongly associated with synovial inflammation showing a direct link between mitochondrial mutations and key proinflammatory pathways.

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Footnotes

  • Funding This study was funded by the Centocor Newman Fellowship, the Health Research Board of Ireland and EU FP6 AutoCure.

  • Competing interest None.

  • Patient consent Informed consent obtained.

  • Ethics approval This study was conducted in accordance with the Declaration of Helsinki and approved by the St Vincent's University Hospital medical research and ethics committee.

  • Provenance and peer review Not commissioned; externally peer reviewed.

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