In this study, we analysed the expression level of sera circulating miRNA-5196 in rheumatoid arthritis (RA) and ankylosing spondylitis (AS) patients before and after tumor necrosis factor (TNF)-α therapy as biomarkers predicting positive treatment outcome. We enrolled 10 RA patients, 13 AS patients, and 12 healthy individuals in the study. The expression of miRNA-5196 was measured by real-time polymerase chain reaction before and after anti-TNF-α therapy. Disease activity of RA patients was assessed using disease activity score 28 (DAS28), whereas ankylosing spondylitis DAS (ASDAS) was used in AS patients. MiRNA-5196 expression was significantly higher in patients with RA and AS before TNF-α therapy than in those following anti-TNF-α therapy and healthy controls. Changes in miRNA-5196 expression positively correlated with delta DAS28 or delta ASDAS, respectively, following TNF-α therapy. In contrast, changes in C-reactive protein (CRP) levels in RA and AS patients did not positively correlate with DAS28 or ASDAS changes. Receiver-operating characteristic analysis showed better diagnostic accuracy of miRNA-5196 expression both in RA (area under curve (AUC) = 0.87, p = 0.055) and AS patients (AUC = 0.90, p = 0.050) compared to CRP levels in RA (AUC = 0.75, p = 0.201) and AS patients (AUC = 0.85, p = 0.086) upon biologic therapy treatment. Finding novel biomarkers, including miRNA-5196 which allow to predict and monitor anti-TNF-α response, would be of clinical value especially during the early phase of RA or AS development.

These results suggest that microRNA-5196 can be used as a potential biomarker characterising SSc. Overall, this study may open new possibilities for the development of microRNA-5196-based diagnostics and therapy in early phases of SSc.

Intratumoral heterogeneity is a driver of breast cancer progression, but the nature of the clonal interactive network involved in this process remains unclear. Here, we optimized the use of optical barcoding to visualize and characterize 31 cancer subclones in vivo. By mapping the clonal composition of thousands of metastases in two clinically relevant sites, the lungs and liver, we found that metastases were highly polyclonal in lungs but not in the liver. Furthermore, the transcriptome of the subclones varied according to their metastatic niche. We also identified a reversible niche-driven signature that was conserved in lung and liver metastases collected during patient autopsies. Among this signature, we found that the tumor necrosis factor–α pathway was up-regulated in lung compared to liver metastases, and inhibition of this pathway affected metastasis diversity. These results highlight that the cellular and molecular heterogeneity observed in metastases is largely dictated by the tumor microenvironment.

The development of therapies that target specific disease subtypes has dramatically improved outcomes for patients with breast cancer. However, survival gains have not been uniform across patients, even within a given molecular subtype. Large collections of publicly available drug screening data matched with transcriptomic measurements have facilitated the development of computational models that predict response to therapy. Here, we generated a series of predictive gene signatures to estimate the sensitivity of breast cancer samples to 90 drugs, comprising FDA-approved drugs or compounds in early development. To achieve this, we used a cell line-based drug screen with matched transcriptomic data to derive in silico models that we validated in large independent datasets obtained from cell lines and patient-derived xenograft (PDX) models. Robust computational signatures were obtained for 28 drugs and used to predict drug efficacy in a set of PDX models. We found that our signature for cisplatin can be used to identify tumors that are likely to respond to this drug, even in absence of the BRCA-1 mutation routinely used to select patients for platinum-based therapies. This clinically relevant observation was confirmed in multiple PDXs. Our study foreshadows an effective delivery approach for precision medicine.

Background and objectivesSystemic sclerosis (SSc) is a chronic multisystem autoimmune disease characterised by skin and  internal organs fibrosis and immune abnormalities. Recent evidence suggests that activated circulating monocytes from SSc patients play a role in SSc pathogenesis due to enhanced expression of tissue inhibitor of metalloproteinases 1 (TIMP-1), IL-8 and reactive oxygen species (ROS) induction, which contribute to fibrosis progression and chronic inflammation. The exact factors driving TIMP-1, IL-8 and ROS secretion are still unknown.The aim of this study was to investigate the expression pattern of profibrotic IL-8, TIMP-1, AP1 transcription factor-Fra2 and ROS induction in peripheral blood monocytes following DZNep (histone methyltransferase inhibitor) and TLR8 agonist (ssRNA) stimulation.Materials and methodsThe expression of Fra2, IL-8 and TIMP-1 and anti-oxidant superoxide dismutase 1 (SOD1) was measured by qRT-PCR in stimulated and unstimulated HC (n = 14) and SSc (n = 17) monocytes. Generation of ROS was determined using luciferase based assay. Fra2 DNA-binding activity was measured in AP-1 transcription assay in monocytic U937 cell line following epigenetic and TLR8 modifications. The level of anti-fibrotic miRNA-5196, which is predicted to bind and inhibit 3’UTR of Fra-2 gene, was also determined in HC and SSc monocytes.ResultsCombination of DZNep+TLR8 enhanced Fra2 (2-fold, p = 0.02), TIMP-1 (2-fold) and IL-8 (7.87-fold, p < 0.001) expression in SSc monocytes. Fra2 DNA-binding activity was 1.5-fold increased upon stimulation. Secreted level of TIMP-1 was 1.46-fold higher in SSc monocytes compared to unstimulated cells. Generated ROS was 2.21-fold (p = 0.0395) higher following DZNep+TLR8 stimulation in monocytic U937 cells. In contrast, miRNA-5196 expression was 2.13-fold decreased in SSc monocytes upon DZNep+TLR8 stimulation. Also the level of SOD1 was decreased in HC and SSc monocytes following stimulation, 2.16-fold (p = 0.025) and 1.56-fold, respectively.ConclusionsThese data suggest that DZNep and TLR8 agonist are able to enhance pro-fibrotic TIMP-1, IL-8 and oxidative stress generation. As opposed by the decrease of anti-fibrotic miRNA-5196 and anti-oxidant SOD1 expression in SSc monocytes, which might be used as a potential modulators of fibrogenesis in SSc.Supported by Homing Plus grant/2013–8/4 from Foundation for Polish Science and UMO-2015/16/S/NZ6/00041 from National Science Centre, Poland.

Background and objectives Several recent studies highlighted the importance of special gene loci related to gout and hyperuricaemia. However, it is remarkable that only a minor fraction of hyperuricemic population produces the symptoms of gout. Discussing the pathomechnism of gouty inflammation a potential autoinflammatory explanation has emerged recently. Induced by MSU crystals, the pathologic activity of a multi-protein complex (NLRP3 inflammasome) might be responsible for the increased activation of interleukin 1-beta (IL-1b), a cytokine playing a central role in inflamation. Toll-like receptors (TLR2-4) have also been studied for the stepped-up production of pre-IL-1b, the pre form of the active metabolit. Human erythrocytes express numerous membrane proteins, several of them already identified as homologue proteins to those with specific function in other tissue, such as urate transporters.Our objective was to verify genetical variations, single nucleotid polymorphisms (SNP) leading to hyperuricemia and gout. We also analysed these gene loci coded proteins linked to elevated urate level, using a unique technique (EryTest) based on the presence of special membrane proteins on human erythrocytes. Materials and methods We studied the results of three groups. Patients with gout, control patients with hyperuricemia but with no arthritic event and patients with normal serum uric acid level. Specific monoclonal antibodies and IgG control were used for quantitative flow cytometry to determine proteins. Polymorphism specific and control primers were used for polymerase chain reaction (PCR) detecting SNPs. Results Increased level of CARD8 polymorphism was found in the gout group compared to the two control groups. Decreased ABCG2 protein expression was detected in heterozygous individuals in gouty and hyperuricemic groups compared to normouricemic controls.Conclusions We presume that depending on complex genetic and environmental factors, patients with CARD8 polymorphisms may have increased inflammatory response and those with decreased ABCG2 protein expression may have higher serum urate levels. From the clinical introduction of EryTest, a simple and rapid clinical method in the evaluation of hyperuricemia is expected. To confirm recent results of genetic variability and the clinical use of EryTest, further investigation is planned. The Fibrous connection A3.1Background and objectives To investigate whether epigenetic changes induced by 3'deazaneplanocin -DZNep and TLR signalling pathway can modulate monocytes to produce tissue-inhibitor of metalloproteinase-1 (TIMP-1) via Fra2 (Fos-related antigen 2) activity, a novel downstream mediator promoting fibrogenesis. Methods AP-1 and TIMP-1 expression following TLR8 treatment was measured by qRT-PCR in monocytes from Systemic sclerosis (SSc) patients (n = 13) and healthy controls (HC) (n = 13). TIMP-1 promoter activity was measured in U397 monocytic cell line using luciferase reporter assay. The effect of DZNep treatment on inhibition of tri-methylation of lysine 27 on h...