A majority of Brazilian patients with rheumatoid arthritis HLA-DRB1 alleles carry both the HLA-DRB1 shared epitope and anti-citrunillated peptide antibodies

The objective of the present study was to evaluate the contribution of the shared epitope (SE), the rheumatoid arthritis (RA) protection model, and the occurrence of anti-cyclic citrullinated peptide (anti-CCP) antibodies in RA patients from a genetically diverse population. One hundred and forty Brazilian RA patients and 161 matched controls were typed for HLA-DRB1 alleles using amplified DNA hybridized with sequence-specific oligonucleotide probes or primers. Patients were stratified according to the presence or absence of SE (DRB1*0401, *0404, *0405, *0101, *1001, and *1402), of the DERAA alleles (DRB1*0103, *0402, *1102, *1103, *1301, *1302, and *1304), and X (all other alleles). Anti-CCP antibodies were measured by ELISA. The combined frequency of SE-positive alleles was significantly greater (76.4 vs 23.6%, P < 0.0001) than the controls. The SE/SE and SE/X genotypes were over-represented (P < 0.0001, OR = 6.02) and DERAA/X was under-represented in RA patients (P < 0.001, OR = 0.49), whereas the frequencies of the SE/DERAA, X/X and X/DERAA genotypes were not significantly different from controls. The frequency of anti-CCP antibodies was higher in SE-positive patients than in SE-negative patients (64.6 vs 44.7%, P = 0.03; OR = 2.25). Although the Brazilian population is highly miscegenated, the results of this study support the findings observed in most genetically homogeneous populations with RA; however, they are not mutually exclusive but rather complementary. The participation of DRB1-DERAA alleles in protection against RA was also observed (OR = 0.4; 95%CI = 0.23-0.68).CNPqFAEP

Lapachol, a compound targeting pyrimidine metabolism, ameliorates experimental autoimmune arthritis

Abstract\ud \ud Background\ud The inhibition of pyrimidine biosynthesis by blocking the dihydroorotate dehydrogenase (DHODH) activity, the prime target of leflunomide (LEF), has been proven to be an effective strategy for rheumatoid arthritis (RA) treatment. However, a considerable proportion of RA patients are refractory to LEF. Here, we investigated lapachol (LAP), a natural naphthoquinone, as a potential DHODH inhibitor and addressed its immunosuppressive properties.\ud \ud \ud Methods\ud Molecular flexible docking studies and bioactivity assays were performed to determine the ability of LAP to interact and inhibit DHODH. In vitro studies were conducted to assess the antiproliferative effect of LAP using isolated lymphocytes. Finally, collagen-induced arthritis (CIA) and antigen-induced arthritis (AIA) models were employed to address the anti-arthritic effects of LAP.\ud \ud \ud Results\ud We found that LAP is a potent DHODH inhibitor which had a remarkable ability to inhibit both human and murine lymphocyte proliferation in vitro. Importantly, uridine supplementation abrogated the antiproliferative effect of LAP, supporting that the pyrimidine metabolic pathway is the target of LAP. In vivo, LAP treatment markedly reduced CIA and AIA progression as evidenced by the reduction in clinical score, articular tissue damage, and inflammation.\ud \ud \ud Conclusions\ud Our findings propose a binding model of interaction and support the ability of LAP to inhibit DHODH, decreasing lymphocyte proliferation and attenuating the severity of experimental autoimmune arthritis. Therefore, LAP could be considered as a potential immunosuppressive lead candidate with potential therapeutic implications for RA.The research leading to these results received funding from the European\ud Union Seventh Framework Programme (FP7-2007-2013) under grant\ud agreement n° HEALTH-F4-2011-281608 (TIMER), from the São Paulo Research\ud Foundation (FAPESP) under grant agreements n° 2009/54014-7, 2011/1967-0,\ud 2012/25075-0, 2014/50265-3, 2012/20990-2 and 2013/08216-2 (Center for\ud Research in Inflammatory Diseases), and from the University of São Paulo\ud NAP-DIN and NPPNS under grant agreement n° 11.1.21625.01.0 and\ud 2012.1.17587.1.1, respectivel

Machine learning techniques for computer-aided classification of active inflammatory sacroiliitis in magnetic resonance imaging

Abstract Background Currently, magnetic resonance imaging (MRI) is used to evaluate active inflammatory sacroiliitis related to axial spondyloarthritis (axSpA). The qualitative and semiquantitative diagnosis performed by expert radiologists and rheumatologists remains subject to significant intrapersonal and interpersonal variation. This encouraged us to use machine-learning methods for this task. Methods In this retrospective study including 56 sacroiliac joint MRI exams, 24 patients had positive and 32 had negative findings for inflammatory sacroiliitis according to the ASAS group criteria. The dataset was randomly split with ~ 80% (46 samples, 20 positive and 26 negative) as training and ~ 20% as external test (10 samples, 4 positive and 6 negative). After manual segmentation of the images by a musculoskeletal radiologist, multiple features were extracted. The classifiers used were the Support Vector Machine, the Multilayer Perceptron (MLP), and the Instance-Based Algorithm, combined with the Relief and Wrapper methods for feature selection. Results Based on 10-fold cross-validation using the training dataset, the MLP classifier obtained the best performance with sensitivity = 100%, specificity = 95.6% and accuracy = 84.7%, using 6 features selected by the Wrapper method. Using the test dataset (external validation) the same MLP classifier obtained sensitivity = 100%, specificity = 66.7% and accuracy = 80%. Conclusions Our results show the potential of machine learning methods to identify SIJ subchondral bone marrow edema in axSpA patients and are promising to aid in the detection of active inflammatory sacroiliitis on MRI STIR sequences. Multilayer Perceptron (MLP) achieved the best results

Lapachol, a compound targeting pyrimidine metabolism, ameliorates experimental autoimmune arthritis

The inhibition of pyrimidine biosynthesis by blocking the dihydroorotate dehydrogenase (DHODH) activity, the prime target of leflunomide (LEF), has been proven to be an effective strategy for rheumatoid arthritis (RA) treatment. However, a considerable proportion of RA patients are refractory to LEF. Here, we investigated lapachol (LAP), a natural naphthoquinone, as a potential DHODH inhibitor and addressed its immunosuppressive properties.Molecular flexible docking studies and bioactivity assays were performed to determine the ability of LAP to interact and inhibit DHODH. In vitro studies were conducted to assess the antiproliferative effect of LAP using isolated lymphocytes. Finally, collagen-induced arthritis (CIA) and antigen-induced arthritis (AIA) models were employed to address the anti-arthritic effects of LAP.We found that LAP is a potent DHODH inhibitor which had a remarkable ability to inhibit both human and murine lymphocyte proliferation in vitro. Importantly, uridine supplementation abrogated the antiproliferative effect of LAP, supporting that the pyrimidine metabolic pathway is the target of LAP. In vivo, LAP treatment markedly reduced CIA and AIA progression as evidenced by the reduction in clinical score, articular tissue damage, and inflammation.Our findings propose a binding model of interaction and support the ability of LAP to inhibit DHODH, decreasing lymphocyte proliferation and attenuating the severity of experimental autoimmune arthritis. Therefore, LAP could be considered as a potential immunosuppressive lead candidate with potential therapeutic implications for RA