The 3'-untranslated region of ADAMTS1 regulates its mRNA stability

ADAMTS1 (a disintegrin and metalloproteinase with thrombospondin motifs 1) is an inflammatory-induced gene. We have previously reported that ADAMTS1 was strongly but transiently expressed in the infarcted heart. In this study, we investigated whether a 3'-untranslated region (UTR) affects the mRNA stability of this gene. When stimulated with tissue necrosis factor (TNF)-alpha, the expression level of ADAMTS1 mRNA rapidly increased, but the induction of ADAMTS1 mRNA peaked at 6h after stimulation, after which the expression levels of ADAMTS1 mRNA decreased. The 3'-UTR ADAMTS1 mRNA contains multiple adenine and uridine-rich elements, suggesting that the 3'-UTR may regulate gene stability. The addition of actinomycin D, an RNA synthesis inhibitor, demonstrated the decay of induced ADAMTS1 mRNA by TNF-alpha. Furthermore, a region containing multiple AUUUA motifs within the ADAMTS1 3'-UTR destabilized transfected Enhanced Green Fluorescence Protein (EGFP) mRNA expression. These results demonstrated that the ADAMTS1 3'-UTR may regulate the expression of ADAMTS1 mRNA.</p

Mechanical strain attenuates cytokine-induced ADAMTS9 expression via transient receptor potential vanilloid type 1

The synovial fluids of patients with osteoarthritis (OA) contain elevated levels of inflammatory cytokines, which induce the expression of a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) and of the matrix metalloproteinase (MMP) in chondrocytes. Mechanical strain has varying effects on organisms depending on the strength, cycle, and duration of the stressor; however, it is unclear under inflammatory stimulation how mechanical strain act on. Here, we show that mechanical strain attenuates inflammatory cytokine-induced expression of matrix-degrading enzymes. Cyclic tensile strain (CTS), as a mechanical stressor, attenuated interleukin (IL)-1β and tumor necrosis factor (TNF)-α-induced mRNA expression of ADAMTS4, ADAMTS9, and MMP-13 in normal chondrocytes (NHAC-kn) and in a chondrocytic cell line (OUMS-27). This effect was abolished by treating cells with mechano-gated channel inhibitors, such as gadolinium, transient receptor potential (TRP) family inhibitor, ruthenium red, and with pharmacological and small interfering RNA-mediated TRPV1 inhibition. Furthermore, nuclear factor κB (NF-κB) translocation from the cytoplasm to the nucleus resulting from cytokine stimulation was also abolished by CTS. These findings suggest that mechanosensors such as the TRPV protein are potential therapeutic targets in treating OA

Osteopontin silencing attenuates bleomycin-induced murine pulmonary fibrosis by regulating epithelial-mesenchymal transition

Idiopathic pulmonary fibrosis (IPF) is the most common and most deadly form of interstitial lung disease. Osteopontin (OPN), a matricellular protein with proinflammatory and profibrotic properties, plays a major role in several fibrotic diseases, including IPF; OPN is highly upregulated in patients' lung samples. In this study, we knocked down OPN in a bleomycin (BLM)-induced pulmonary fibrosis (PF) mouse model using small interfering RNA (siRNA) to determine whether the use of OPN siRNA is an effective therapeutic strategy for IPF. We found that fibrosing areas were significantly smaller in specimens from OPN siRNA-treated mice. The number of alveolar macrophages, neutrophils, and lymphocytes in bronchoalveolar lavage fluid was also reduced in OPN siRNA-treated mice. Regarding the expression of epithelial-mesenchymal transition (EMT)-related proteins, the administration of OPN-siRNA to BLM-treated mice upregulated E-cadherin expression and downregulated vimentin expression. Moreover, in vitro, we incubated the human alveolar adenocarcinoma cell line A549 with transforming growth factor (TGF)-beta 1 and subsequently transfected the cells with OPN siRNA. We found a significant upregulation of Col1A1, fibronectin, and vimentin after TGF-beta 1 stimulation in A549 cells. In contrast, a downregulation of Col1A1, fibronectin, and vimentin mRNA levels was observed in TGF-beta 1-stimulated OPN knockdown A549 cells. Therefore, the downregulation of OPN effectively reduced pulmonary fibrotic and EMT changes both in vitro and in vivo. Altogether, our results indicate that OPN siRNA exerts a protective effect on BLM-induced PF in mice. Our results provide a basis for the development of novel targeted therapeutic strategies for IPF

Potential of a Novel Chemical Compound Targeting Matrix Metalloprotease-13 for Early Osteoarthritis: An In Vitro Study

Osteoarthritis is a progressive disease characterized by cartilage destruction in the joints. Matrix metalloproteinases (MMPs) and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTSs) play key roles in osteoarthritis progression. In this study, we screened a chemical compound library to identify new drug candidates that target MMP and ADAMTS using a cytokine-stimulated OUMS-27 chondrosarcoma cells. By screening PCR-based mRNA expression, we selected 2-(8-methoxy-2-methyl-4-oxoquinolin-1(4H)-yl)-N-(3-methoxyphenyl) acetamide as a potential candidate. We found that 2-(8-methoxy-2-methyl-4-oxoquinolin-1(4H)-yl)-N-(3-methoxyphenyl) acetamide attenuated IL-1 beta-induced MMP13 mRNA expression in a dose-dependent manner, without causing serious cytotoxicity. Signaling pathway analysis revealed that 2-(8-methoxy-2-methyl-4-oxoquinolin-1(4H)-yl)-N-(3-methoxyphenyl) acetamide attenuated ERK- and p-38-phosphorylation as well as JNK phosphorylation. We then examined the additive effect of 2-(8-methoxy-2-methyl-4-oxoquinolin-1(4H)-yl)-N-(3-methoxyphenyl) acetamide in combination with low-dose betamethasone on IL-1 beta-stimulated cells. Combined treatment with 2-(8-methoxy-2-methyl-4-oxoquinolin-1(4H)-yl)-N-(3-methoxyphenyl) acetamide and betamethasone significantly attenuated MMP13 and ADAMTS9 mRNA expression. In conclusion, we identified a potential compound of interest that may help attenuate matrix-degrading enzymes in the early osteoarthritis-affected joints

Tumor growth inhibitory effect of ADAMTS1 is accompanied by the inhibition of tumor angiogenesis

Angiogenesis plays an important role in tumor progression. Several reports have demonstrated that a disintegrin and metalloproteinase with thrombospondin motifs1 (ADAMTS1) inhibited angiogenesis via multiple mechanisms. The aim of this study was to investigate the effect of ADAMTS1 on endothelial cells in vitro and on tumor growth with regard to angiogenesis in vivo. We examined the effects of the transfection of ADAMTS1 using two constructs, full-length ADAMTS1 (full ADAMTS1) and catalytic domain-deleted ADAMTS1 (delta ADAMTS1). Transfection of both the full ADAMTS1 and delta ADAMTS1 gene constructs demonstrated the secretion of tagged-ADAMTS1 protein into the conditioned medium, so we examined the effects of ADAMTS1-containing conditioned medium on endothelial cells. Both types of conditioned media inhibited endothelial tube formation, and this effect was completely abolished after immunoprecipitation of the secreted protein from the medium. Both types of conditioned media also inhibited endothelial cell migration and proliferation. We then examined the impact of ADAMTS1 on endothelial cell apoptosis. Both conditioned media increased the number of Annexin V-positive endothelial cells and caspase-3 activity and this effect was attenuated when z-vad was added. These results indicated that ADAMTS1 induced endothelial cell apoptosis. We next examined the effects of ADAMTS1 gene transfer into tumor-bearing mice. Both full ADAMTS1 and delta ADAMTS1 significantly inhibited the subcutaneous tumor growth. Collectively, our results demonstrated that ADAMTS1 gene transfer inhibited angiogenesis in vitro and in vivo, likely as a result of the induction of endothelial cell apoptosis by ADAMTS1 that occurs independent of the protease activity

Association between rs11362 polymorphism in the beta-defensin 1 (DEFB1) gene and dental caries: A meta-analysis

Hatipoglu, Omer/0000-0002-4628-8551WOS: 000572657700008PubMed: 32603779Objectives: Beta-defensin 1, encoded by the DEFB1 gene, is an important molecule that confers protection from dental caries. Numerous studies have been conducted on the rs11362 polymorphism in the DEFB1 gene. We evaluated the results from studies that have investigated the association between rs11362 polymorphism and dental caries, through a meta-analysis. Methods: This meta-analysis was designed according to the PRISMA statement guideline. Electronic databases (PubMed, Web of Science, Scopus, and Cochrane Library) were scanned by two independent researchers. the publication bias was determined by statistical analyses using funnel plot, Egger regression test, and Begg and Mazumdar rank correlation test. Heterogeneity was evaluated using the chi-square test, tau-square, and Higgins I-2 test. Odds ratio (OR) was used to measure the effect size. Results: Rank correlation and regression procedures showed the absence of publication bias in the meta-analysis (p > 0.05). the DEFB1 rs11362 polymorphism in the heterozygous (CC vs. CT: OR = 2.20, 95% confidence interval (CI): 1.17, 4.10; p = 0.014) and dominant (CC vs. CT + TT: OR = 3.11, 95% CI: 1.18, 8.21; p = 0.022) models in the permanent dentition subgroup showed significant differences. However, there was no significant difference between any model in either the deciduous dentition (p > 0.05) or the mixed dentition subgroups (p > 0.05). Conclusions: This meta-analysis suggests that the DEFB1 rs11362 polymorphism is associated with dental caries in permanent dentition. Moreover, individuals with the TT genotype were found to have seven times higher risk of dental caries than individuals with the CC genotype. There was no such association or statistical difference observed for deciduous and mixed dentitions. (C) 2020 Japanese Association for Oral Biology. Published by Elsevier B.V. All rights reserved

Effects of the carbonic anhydrase VI gene polymorphisms on dental caries: A meta-analysis

Hatipoglu, Omer/0000-0002-4628-8551WOS: 000507597600010PubMed: 31895503Background. Carbonic anhydrase VI (CA VI) is considered to greatly participate in the buffering of saliva, ion transport, the regulation of pH, secretory processes, and saliva production. Various studies have been conducted to investigate the relationship between CA VI and dental caries. Objectives. the goal of this study was to make a meta-analysis of studies that examined the effects of the CA VI gene polymorphisms on dental caries. Material and methods. the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement guide was followed. Electronic databases (PubMed, Web of Science, Scopus, and Cochrane Library) were scanned by 2 independent researchers. the funnel plot, Egger's regression and Begg and Mazumdar's rank correlation test were used to determine publication bias. Cohen's d was used to measure the effect size. Results. Four studies were included in the meta-analysis; a total of 3 polymorphisms (rs2274327, rs2274328, rs2274333) and a total of 13 polymorphism models were analyzed. According to Egger's regression and the Begg and Mazumdar's test, the meta-analysis had no significant publication bias (p > 0.05). the highest susceptibility effect was noticed in the rs2274328 (AA vs CC) model (d = 0.18; 95% CI (confidence interval): -1.77, 2.13), but this effect was not significant (p = 0.237), and the highest protective effect was observed in the rs2274328 (AA vs AC) model (d = -0.13, 95% CI: -1.36, 1.11), but this effect was not significant, either (p = 0.195). No association was found between any of the polymorphism models and dental caries (p > 0.05). Conclusions. Even though CA VI plays an important role in the buffering of saliva, it was shown that polymorphisms in the CA VI gene did not affect the process of dental caries