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Monomeric C reactive protein (mCRP) regulates inflammatory responses in human and mouse chondrocytes

Laboratory Investigation (2021)Cite this article

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Abstract

C-reactive protein (CRP) is an acute-phase protein that is used as an established biomarker to follow disease severity and progression in a plethora of inflammatory diseases. However, its pathophysiologic mechanisms of action are still poorly defined and remain elusive. CRP, in its pentameric form, exhibits weak anti-inflammatory activity. On the contrary, the monomeric isoform (mCRP) exhibits potent pro-inflammatory properties in endothelial cells, leukocytes, and platelets. So far, no data exists regarding mCRP effects in human or mouse chondrocytes. This work aimed to verify the pathophysiological relevance of mCRP in the etiology and/or progression of osteoarthritis (OA). We investigated the effects of mCRP in cultured human primary chondrocytes and in the chondrogenic ATDC5 mouse cell line. We determined mRNA and protein levels of relevant factors involved in inflammatory responses and the modulation of nitric oxide synthase type II (NOS2), an early inflammatory molecular target. We demonstrate, for the first time, that monomeric C reactive protein increases NOS2, COX2, MMP13, VCAM1, IL-6, IL-8, and LCN2 expression in human and murine chondrocytes. We also demonstrated that NF-kB is a key factor in the intracellular signaling of mCRP-driven induction of pro-inflammatory and catabolic mediators in chondrocytes. We concluded that mCRP exerts a sustained catabolic effect on human and murine chondrocytes, increasing the expression of inflammatory mediators and proteolytic enzymes, which can promote extracellular matrix (ECM) breakdown in healthy and OA cartilage. In addition, our results implicate the NF-kB signaling pathway in catabolic effects mediated by mCRP.

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Fig. 1: mCRP pro-inflammatory effect on human primary osteoarthritis chondrocytes.
Fig. 2: mCRP pro-inflammatory effect on human primary healthy chondrocytes.
Fig. 3: mCRP pro-inflammatory effect on murine chondrocyte cell line.
Fig. 4: Nf-κβ role in mCRP signaling: Nf-κβ inhibition with PDTC in murine chondrocyte cell line.

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Acknowledgements

OG and FL are Staff Personnel (I3SNS stable Researcher) of Xunta de Galicia (Servizo Galego de Saude, SERGAS) through a research-staff contract (ISCIII/SERGAS). VF is currently a “Sara Borrell” Researcher funded by ISCIII and FEDER (CD16/00111). RG and JC are “Miguel Servet” Researchers funded by Instituto de Salud Carlos III (ISCIII) and FEDER. CRF is a pre-doctoral research scholar funded by ISCIII and FEDER (Exp.18/00188). OG, MAGG, and RG are members of RETICS Programme, RD16/0012/0014 (RIER: Red de Investigación en Inflamación y Enfermedades Reumáticas) via Instituto de Salud Carlos III (ISCIII) and FEDER. FL is a member of CIBERCV (Centro de Investigación Biomédica en Red de Enfermedades Cardiovasculares). The work of OG and JP (PI17/00409), RG (PI16/01870) and FL (PI18/00821 and CB16/11/00226) was funded by Instituto de Salud Carlos III and FEDER. OG is a beneficiary of a project funded by Research Executive Agency of the European Union in the framework of MSCA-RISE Action of the H2020 Programme (Project number 734899). OG is beneficiary of a grant funded by Xunta de Galicia, Consellería de Educación, Universidade e Formación Profesional and Consellería de Economía, Emprego e Industria (GAIN), GPC IN607B2019/10.

Funding

The funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the manuscript.

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Affiliations

  1. SERGAS (Servizo Galego de Saude) and IDIS (Instituto de Investigación Sanitaria de Santiago), NEIRID Lab (Neuroendocrine Interactions in Rheumatology and Inflammatory Diseases), Research Laboratory 9, Santiago University Clinical Hospital, Santiago de Compostela, Spain

    Clara Ruiz-Fernández, María Gonzalez-Rodríguez, Vera Francisco, Javier Conde, Jesús Pino & Oreste Gualillo

  2. International PhD School of the University of Santiago de Compostela (EDIUS), Doctoral Programme in Medicine Clinical Research, Santiago de Compostela, Spain

    Clara Ruiz-Fernández

  3. Roosevelt University, College of Pharmacy, Schaumburg, IL, USA

    Ibraheem M. Rajab & Lawrence A. Potempa

  4. Musculoskeletal Pathology Laboratory, IDIS, Santiago University Clinical Hospital, Santiago de Compostela, Spain

    Rodolfo Gómez

  5. Molecular and Cellular Cardiology Group, SERGAS (Servizo Galego de Saude) and IDIS (Instituto de Investigación Sanitaria de Santiago), Research Laboratory 7, Santiago University Clinical Hospital, Santiago de Compostela, Spain

    Francisca Lago

  6. Research Unit of Medical Imaging, Physics, and Technology, Faculty of Medicine, University of Oulu, Oulu, Finland

    Ali Mobasheri

  7. Department of Regenerative Medicine, State Research Institute Centre for Innovative Medicine, Vilnius, Lithuania

    Ali Mobasheri

  8. University Medical Center Utrecht, Departments of Orthopedics, Rheumatology and Clinical Immunology, Utrecht, The Netherlands

    Ali Mobasheri

  9. Hospital Universitario Marqués de Valdecilla, Epidemiology, Genetics and Atherosclerosis Research Group on Systemic Inflammatory Diseases, IDIVAL, University of Cantabria, Avenida de Valdecilla s/n, Santander, Cantabria, Spain

    Miguel Angel Gonzalez-Gay

  10. SERGAS, Santiago University Clinical Hospital, Division of Rheumatology, Santiago de Compostela, Spain

    Antonio Mera

Authors
  1. Clara Ruiz-Fernández
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  2. María Gonzalez-Rodríguez
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  3. Vera Francisco
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Contributions

CRF, MG, and VF participated in acquisition of data, analysis, and interpretation of data and critical revision of the manuscript. AM, RG, JC, FL, JP, MAGG, and AM participated in the acquisition of data and samples, drafting of the manuscript, and statistical analysis. IMR participated in C-rmCRP preparation and editing the manuscript. LAP participated in the analysis and interpretation of data and drafting/editing the manuscript. OG participated in the conception and design of the study, in the analysis and interpretation of data, critical revision of the manuscript, and scientific supervision of experiments.

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Correspondence to Oreste Gualillo.

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Ruiz-Fernández, C., Gonzalez-Rodríguez, M., Francisco, V. et al. Monomeric C reactive protein (mCRP) regulates inflammatory responses in human and mouse chondrocytes. Lab Invest (2021). https://doi.org/10.1038/s41374-021-00584-8

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