Dysregulated circadian rhythm pathway in human osteoarthritis: NR1D1 and BMAL1 suppression alters TGF-β signaling in chondrocytes
Autor: | R. Akagi, Oscar Alvarez-Garcia, Kathleen M. Fisch, Andrew I. Su, Y. Muramatsu, Martin Lotz, Yorikazu Akatsu, Takahisa Sasho, Masahiko Saito, Takeshi Teramura |
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Rok vydání: | 2016 |
Předmět: |
0301 basic medicine
Adult Cartilage Articular Male endocrine system medicine.medical_specialty Small interfering RNA Adolescent Biomedical Engineering Chondrocyte Article 03 medical and health sciences Young Adult 0302 clinical medicine Chondrocytes Rheumatology Transforming Growth Factor beta Internal medicine medicine Humans Orthopedics and Sports Medicine RNA Messenger Regulation of gene expression Messenger RNA biology Cartilage homeostasis Cartilage RNA ARNTL Transcription Factors Transforming growth factor beta Middle Aged Osteoarthritis Knee Cell biology Circadian Rhythm 030104 developmental biology Endocrinology medicine.anatomical_structure Gene Expression Regulation Gene Knockdown Techniques Nuclear Receptor Subfamily 1 Group D Member 1 biology.protein Female 030217 neurology & neurosurgery Signal Transduction |
Zdroj: | Osteoarthritis and cartilage. 25(6) |
ISSN: | 1522-9653 |
Popis: | Summary Objectives Circadian rhythm (CR) was identified by RNA sequencing as the most dysregulated pathway in human osteoarthritis (OA) in articular cartilage. This study examined circadian rhythmicity in cultured chondrocytes and the role of the CR genes NR1D1 and BMAL1 in regulating chondrocyte functions. Methods RNA was extracted from normal and OA-affected human knee cartilage ( n = 14 each). Expression levels of NR1D1 and BMAL1 mRNA and protein were assessed by quantitative PCR and immunohistochemistry. Human chondrocytes were synchronized and harvested at regular intervals to examine circadian rhythmicity in RNA and protein expression. Chondrocytes were treated with small interfering RNA (siRNA) for NR1D1 or BMAL1, followed by RNA sequencing and analysis of the effects on the transforming growth factor beta (TGF-β) pathway. Results NR1D1 and BMAL1 mRNA and protein levels were significantly reduced in OA compared to normal cartilage. In cultured human chondrocytes, a clear circadian rhythmicity was observed for NR1D1 and BMAL1. Increased BMAL1 expression was observed after knocking down NR1D1, and decreased NR1D1 levels were observed after knocking down BMAL1. Sequencing of RNA from chondrocytes treated with NR1D1 or BMAL1 siRNA identified 330 and 68 significantly different genes, respectively, and this predominantly affected the TGF-β signaling pathway. Conclusions The CR pathway is dysregulated in OA cartilage. Interference with circadian rhythmicity in cultured chondrocytes affects TGF-β signaling, which is a central pathway in cartilage homeostasis. |
Databáze: | OpenAIRE |
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