Circadian Timekeeping Is Disturbed in Rheumatoid Arthritis at Molecular Level

Vesa-Petteri Kouri,Juri Olkkonen,Emilia Kaivosoja,Jami Mandelin,Yrjö T Konttinen,Iiris Hovatta,Mari Ainola,Juuso Juhila,Oliver Frey

doi:10.1371/journal.pone.0054049

Abstract

IntroductionPatients with rheumatoid arthritis (RA) have disturbances in the hypothalamic-pituitary-adrenal (HPA) axis. These are reflected in altered circadian rhythm of circulating serum cortisol, melatonin and IL-6 levels and in chronic fatigue. We hypothesized that the molecular machinery responsible for the circadian timekeeping is perturbed in RA. The aim of this study was to investigate the expression of circadian clock in RA.MethodsGene expression of thirteen clock genes was analyzed in the synovial membrane of RA and control osteoarthritis (OA) patients. BMAL1 protein was detected using immunohistochemistry. Cell autonomous clock oscillation was started in RA and OA synovial fibroblasts using serum shock. The effect of pro-inflammatory stimulus on clock gene expression in synovial fibroblasts was studied using IL-6 and TNF-α.ResultsGene expression analysis disclosed disconcerted circadian timekeeping and immunohistochemistry revealed strong cytoplasmic localization of BMAL1 in RA patients. Perturbed circadian timekeeping is at least in part inflammation independent and cell autonomous, because RA synovial fibroblasts display altered circadian expression of several clock components, and perturbed circadian production of IL-6 and IL-1β after clock resetting. However, inflammatory stimulus disturbs the rhythm in cultured fibroblasts. Throughout the experiments ARNTL2 and NPAS2 appeared to be the most affected clock genes in human immune-inflammatory conditions.ConclusionWe conclude that the molecular machinery controlling the circadian rhythm is disturbed in RA patients.

Highlights

Patients with rheumatoid arthritis (RA) have disturbances in the hypothalamic-pituitary-adrenal (HPA) axis
We conclude that the molecular machinery controlling the circadian rhythm is disturbed in RA patients
Clock Gene Expression is Disturbed in RA Synovium Driven by our hypothesis that the circadian timekeeping is perturbed in RA, we analyzed expression levels of clock genes in the synovial membrane of RA patients in comparison to OA patients

Summary

Introduction

Patients with rheumatoid arthritis (RA) have disturbances in the hypothalamic-pituitary-adrenal (HPA) axis. These are reflected in altered circadian rhythm of circulating serum cortisol, melatonin and IL-6 levels and in chronic fatigue. We hypothesized that the molecular machinery responsible for the circadian timekeeping is perturbed in RA. The aim of this study was to investigate the expression of circadian clock in RA. Endogenous circadian rhythm is maintained by the central circadian timekeeper, the suprachiasmatic nuclei of the hypothalamus. The clock machinery is composed of heterodimeric transcription factors and transcription regulatory proteins. The expression of these factors oscillates rhythmically over 24-hour cycles [1,2,3]. Altered circadian rhythm can cause e.g., cancer and obesity [5,6]

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