The transcriptional response of normal and rheumatoid arthritis synovial fibroblasts to hypoxia

Rey, Manuel; Izquierdo, Elena; Usategui, Alicia; Gonzalo, Elena Sierra; Blanco, Francisco J.; Acquadro, Francesco; Pablos, José L.

doi:10.1002/art.27750

Cited by 50 publications

(36 citation statements)

References 68 publications

(58 reference statements)

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“…The cellular response to hypoxia is mainly driven by the transcription factor HIF [25]. Thus, in this study, measurement of HIF-1α protein in the presence of HMGB1 was considered as a marker of hypoxia.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of HMGB1-Induced Angiogenesis by Cilostazol via SIRT1 Activation in Synovial Fibroblasts from Rheumatoid Arthritis

et al. 2014

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High mobility group box chromosomal protein 1 (HMGB-1) released from injured cells plays an important role in the development of arthritis. This study investigated the anti-angiogenic effects of cilostazol in collagen-induced arthritis (CIA) of mice, and the underlying mechanisms involved. The expressions of HIF-1α, VEGF, NF-κB p65 and SIRT1 in synovial fibroblasts obtained from rheumatoid arthritis (RA) patients were assessed by Western blotting, and in vitro and in vivo angiogenesis were analyzed. Tube formations by human microvascular endothelial cells (HMVECs) were significantly increased by direct exposure to HMGB1 or to conditioned medium derived from HMGB1-stimulated RA fibroblasts, and these increases were attenuated by cilostazol, the latter of which was blocked by sirtinol. HMGB1 increased the expression of HIF-1α and VEGF and concomitantly increased nuclear NF-κB p65 and DNA binding activity, but these effects of HMGB1 were inhibited by cilostazol. SIRT1 protein expression was time-dependently decreased (3–24 hr) by HMGB1, which was recovered by pretreatment with cilostazol (1–30 µM) or resveratrol, accompanying with increased SIRT1 deacetylase activity. In the tibiotarsal joint tissues of CIA mice treated with vehicle, HIF-1α- and VEGF-positive spots and CD31 staining were markedly exaggerated, whereas SIRT1 immunofluorescence was diminished. These variables were wholly reversed in cilostazol (30 mg/kg/day)-treated mice. Furthermore, number of blood vessels stained by von Willebrand factor antibody was significantly lower in cilostazol-treated CIA mice. Summarizing, cilostazol activated SIRT1 and inhibited NF-κB-mediated transcription, thereby suppressing the expression of HIF-1α and VEGF. In addition, cilostazol caused HIF-1α deacetylation by enhancing SIRT1 activity and reduced VEGF production, thereby had an anti-angiogenic effect in vitro studies and in CIA murine model.

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Section: Discussionmentioning

confidence: 99%

Inhibition of HMGB1-Induced Angiogenesis by Cilostazol via SIRT1 Activation in Synovial Fibroblasts from Rheumatoid Arthritis

et al. 2014

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“…Previous investigations have demonstrated very low levels of HIF-1α protein under normal O 2 conditions in transformed cell lines 19, 20 . For the purpose of evaluating the potential role of HIF-1α activity in normoxia, we analyzed the mRNA expression of target genes known to be regulated by HIF-1α in hypoxia 21, 22 . In addition to the strong down-regulation of HIF-1α mRNA (83.2 ± 4%, mean ± SEM), we found a considerable down-regulation of HIF-1α-regulated genes such as pyruvate dehydrogenase kinase 1 (PDK1) (60.4 ± 2.8%, mean ± SEM), GAPDH (29 ± 10.4%, mean ± SEM), apelin (APLN) (60.6 ± 6%, mean ± SEM) and inhibin beta (INHBB) (52.9 ± 6.9%, mean ± SEM).…”

Section: Resultsmentioning

confidence: 99%

Hif-1α Knockdown Reduces Glycolytic Metabolism and Induces Cell Death of Human Synovial Fibroblasts Under Normoxic Conditions

Rey

Valín

Usategui

et al. 2017

Sci Rep

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Increased glycolysis and HIF-1α activity are characteristics of cells under hypoxic or inflammatory conditions. Besides, in normal O2 environments, elevated rates of glycolysis support critical cellular mechanisms such as cell survival. The purpose of this study was to analyze the contribution of HIF-1α to the energy metabolism and survival of human synovial fibroblasts (SF) under normoxic conditions. HIF-1α was silenced using lentiviral vectors or small-interfering RNA (siRNA) duplexes. Expression analysis by qRT-PCR and western blot of known HIF-1α target genes in hypoxia demonstrated the presence of functional HIF-1α in normoxic SF and confirmed the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a HIF-1α target even in normoxia. HIF-1α silencing induced apoptotic cell death in cultured SF and, similarly, treatment with glycolytic, but not with OXPHOS inhibitors, induced SF death. Finally, in vivo HIF-1α targeting by siRNA showed a significant reduction in the viability of human SF engrafted into a murine air pouch. Our results demonstrate that SF are highly dependent on glycolytic metabolism and that HIF-1α plays a regulatory role in glycolysis even under aerobic conditions. Local targeting of HIF-1α provides a feasible strategy to reduce SF hyperplasia in chronic arthritic diseases.

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“…The main mechanism that promotes the development of new vessel formation is hypoxia [97, 98]. Hypoxic state leads to the activation of HIF, which activates the expression of HIF-responsive genes, including vascular endothelial growth factor (VEGF), which are important in synovium angiogenic processes and RA perpetuation [99].…”

Section: Metabolic Changes Deregulate Fls Phenotype After Activationmentioning

confidence: 99%

Fibroblast-like synoviocyte metabolism in the pathogenesis of rheumatoid arthritis

et al. 2017

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An increasing number of studies show how changes in intracellular metabolic pathways alter tumor and immune cell function. However, little information about metabolic changes in other cell types, including synovial fibroblasts, is available. In rheumatoid arthritis (RA), fibroblast-like synoviocytes (FLS) are the most common cell type at the pannus–cartilage junction and contribute to joint destruction through their production of cytokines, chemokines, and matrix-degrading molecules and by migrating and invading joint cartilage. In this review, we show that these cells differ from healthy synovial fibroblasts, not only in their marker expression, proto-oncogene expression, or their epigenetic changes, but also in their intracellular metabolism. These metabolic changes must occur due to the stressful microenvironment of inflamed tissues, where concentrations of crucial nutrients such as glucose, glutamine, and oxygen are spatially and temporally heterogeneous. In addition, these metabolic changes will increase metabolite exchange between fibroblast and other synovial cells, which can potentially be activated. Glucose and phospholipid metabolism as well as bioactive lipids, including sphingosine-1-phosphate and lysophosphatidic acid, among others, are involved in FLS activation. These metabolic changes likely contribute to FLS involvement in aspects of immune response initiation or abnormal immune responses and strongly contribute to joint destruction.

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The transcriptional response of normal and rheumatoid arthritis synovial fibroblasts to hypoxia

Cited by 50 publications

References 68 publications

Inhibition of HMGB1-Induced Angiogenesis by Cilostazol via SIRT1 Activation in Synovial Fibroblasts from Rheumatoid Arthritis

Inhibition of HMGB1-Induced Angiogenesis by Cilostazol via SIRT1 Activation in Synovial Fibroblasts from Rheumatoid Arthritis

Hif-1α Knockdown Reduces Glycolytic Metabolism and Induces Cell Death of Human Synovial Fibroblasts Under Normoxic Conditions

Fibroblast-like synoviocyte metabolism in the pathogenesis of rheumatoid arthritis

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