Search

Background: Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by excess morbidity and mortality from cardiovascular disease. In addition, metabolic alterations have been observed in these patients, which significantly contributes to the cardiovascular risk burden. Thus, the identification of therapies able to mitigate the cardiometabolic alterations in RA is essential. In addition, numerous studies suggest that cardiometabolic risks are mediated through adipocytokines. However, this relationship is not completely defined in RA. Objectives: 1) To evaluate the relationship among cardiometabolic risk factors and the levels of adipocytokines and autoantibodies in RA patients. 2) To analyze the effects anti-TNFα and anti-IL6 therapies on the cardiometabolic alterations. Methods: 1.- A cross-sectional study including 100 RA patients and 50 age-matched healthy donors was carried out. Different parameters related to the cardiometabolic risk were analyzed, including: lipid profile, atherogenic index, ratio ApoB/ApoA, insulin resistance (IR), obesity, hypertension, and the SCORE. Levels of adipocytokines (TNFα, IL6, IL1β, visfatin, adiponectin, leptin and resistin) were evaluated in serum. Carotid intima media thickness (CIMT) was evaluated as atherosclerosis marker. 2.-A prospective study in 15 RA patients before and after 3 months of anti-TNFα therapy and 15 RA patients before and after 3 months of treatment with tocilizumab (TCZ) was performed. All the parameters evaluated in the cross-sectional cohort were tested in this prospective study. Results: RA patients had elevated levels of leptin/adiponectin ratio, visfatin, resistin and inflammatory markers in serum. Our cohort of RA patients displayed increased rates of cardiometabolic risk factors, such as insulin resistance, hypertension, SCORE, pathologic CIMT, atherogenic index and ratio apoB/apoA. The alteration in the levels of adipocytokines were closely related to the autoimmunity, disease activity and clinical inflammatory markers. Of note, visfatin and C3 complement levels were determinant for IR, high levels of SCORE, increased parameters of CVD risk defined by apoB/apoA ratio and pathologic CIMT. Both biological therapies reduced clinical inflammatory markers and disease activity after 3 months of treatment. Anti-TNFα therapy modulated the cytoadipokine profile, reducing serum levels of IL-6, IL-1β, resistin and visfatin, decreasing IR. After treatment with tocilizumab, serum levels of C3 complement, IL-1β and resistin were reduced. Conclusion: 1) Altered adipocytokine profile is closely related to the increased cardiometabolic risk factors associated with RA, where autoimmunity and systemic inflammation play a key role. 2) Anti-TNFα and anti-IL6R therapies, administered for 3 months, could have beneficial effects in the reduction of cardiometabolic risk factors in RA. Acknowledgement: Funded by ISCIII (PI18/00837, CP15/00158, PI17/01316 and RIER RD16/0012/0015) co-funded with FEDER Disclosure of Interests: None declared

Objectives 1) To compare the real-world effectiveness among RA patients starting 3 biologic DMARDs (bDMARDs). 2) To evaluate whether patients’ clinical characteristics influence the response to specific bDMARDs. 3) To analyze the impact of bDMARDs on inflammatory profiles. Methods Eighty RA patients initiating TNFi, 22 patients treated with TCZ and 28 treated with RTX, the last ones after switching from TNFi and given as second bDMARDs, were included in the study. Effectiveness was assessed before and after 3 and 6 months of treatment by the European League Against Rheumatism (EULAR) criteria. Adjusted differences were estimated with multivariable linear regression. Differences in treatment outcomes were calculated as a function of patients’ characteristics, through regression modelling. Extensive analysis of the inflammatory profile using a multiplex were also performed. Results A high proportion of patients starting TNFi showed early EULAR Good Response (65,8% responders at 3 months). After 6 months of therapy this percentage even increased (70,6%), so that tender joints’ counts, swollen joints count, SDAI and HAQ improved significantly. After switching from a first TNFi, RTX treatment promoted better early response (81,8% responders after 3 months) than TCZ (67% responders). Differences remained significant after adjusting for potential confounders. Yet, after 6 months, efficacy was similar among both non-TNFi DMARDs, even improving the effectiveness in TCZ-treated RA patients based on EULAR response rate (DAS28 change: 5.09±1.4 to 2.7±1,17) vs RTX-treated patients (DAS28 change: 4.8±1.3 to 3-1±1.3). Patients older, and those suffering arterial hypertension (AH) or hyperlipidemia showed less effectiveness for both, TNFi and non-TNFi DMARDs, most remarkable for RTX and after 6 months of treatment. Analysis of the inflammatory profile, by evaluating 27 key biomarkers of inflammation, showed, in patients’ responders to therapy, specific profiles depending on the biologic DMARD administered. Thus, after 6 months of RTX, a generalized reduction in the expression levels of 22/27 proteins evaluated was observed, while TCZ mainly inhibited proteins involved in cell maturation, proliferation and leukocytes recruitment (IL6, GCSF, IP-10, IFNγ, RANTES). On the other hand, TNFi mainly reduced molecules related to leukocytes’ recruitment (IL-2, IL-8, MIP1β, RANTES). Conclusion 1) Treatment outcomes among RA patients treated with non-TNFi bDMARDs in standard clinical practice, are in line with a superior early effectiveness of RTX compared with TCZ. However, effectiveness in TCZ-treated RA patients increases after 6 months of treatment vs RTX. 2) Patients characteristics influence the response to bDMARDs, suggesting the need of evaluating the benefit risk/ratio based on a personalized clinical evaluation. 3) Anti-inflammatory benefits associated with bDMARDs treatments can differ despite similar drug efficacy against RA, indicating that drug-specific mechanisms are involved in regulating these processes. Acknowledgement Funded by PI-0285-2017, ISCIII, PI15/00837 and RIER RD16/0012/0015, co-funded with FEDER Disclosure of Interests Chary Lopez-Pedrera: None declared, Nuria Barbarroja Puerto: None declared, Carlos Perez-Sanchez: None declared, Alejandro Ibanez-Costa: None declared, Maria Luque-Tevar: None declared, Alejandra Patino-Trives: None declared, Ivan Arias de la Rosa: None declared, Maria del Carmen Abalos-Aguilera: None declared, Rafaela Ortega Castro: None declared, Alejandro Escudero Contreras: None declared, Perez Sanchez Laura: None declared, Carlos Rodriguez-Escalera: None declared, Dolores Ruiz-Montesinos: None declared, Carmen Maria Romero-Barco: None declared, Natalia Mena-Vazquez: None declared, Julia Uceda Speakers bureau: abbie, pfizer, novartis, janmsen, Ma Dolores Toledo-Coello: None declared, M angeles Aguirre-Zamorano: None declared, Eduardo Collantes Estevez: None declared

Objectives: 1. To characterize the mRNAs and microRNAs transcriptomes of monocytes from systemic lupus erythematosus (SLE) patients and their association with the pathophysiology of the disease. 2. To evaluate the role of anti-dsDNA antibodies in the regulation of these processes. 3. To investigate the molecular mechanisms involved in the efficacy of Fluvastatin in preventing the atherothrombotic risk. Methods: Monocytes from peripheral blood of 81 SLE patients and 40 healthy donors (HD) were purified by negative immunomagnetic selection. Then, gene expression microarray (Agilent G4112F platform) and nCounter microRNA expression arrays (Nanostring) were performed. Functional categorization of altered genes and miRNAs was made using IPA software, and interaction networks were identified. Genes and miRNAs integrating the networks were validated in the whole cohorts by RT-PCR. Predicted miRNA-mRNA interactions were tested by microRNA over-expression or inhibition experiments. Serum and cellular inflammatory and oxidative profiles were evaluated by multiplex assay, PCR and specific commercial kits, respectively; phosphorylation status of intracellular proteins was analyzed by PathScan array. To evaluate the clinical significance of the parameters analyzed, correlation and association studies were performed. Mechanistic studies were developed to typify the specific effects of the anti-dsDNA antibodies on monocytes. Besides, the beneficial effects of ex vivo Fluvastatin treatment on the monocyte molecular profiles were assessed. Results: Microarray identified 553 altered genes in SLE monocytes. Relevant biofunctions and disorders on which these genes were involved included inflammatory, immunological, cardiovascular, neurological, renal and reproductive disease. Analysis of miRNA profiles showed altered expression of 35 miRNAs in SLE monocytes. Sixty-one genes were inversely correlated and predicted as CVD-related target genes of 26 differentially expressed miRNAs. Transfection studies confirmed the relationship between specific miRNAs and their identified target genes. Association of these genes and miRNAs with the anti-dsDNA positivity, early atherosclerosis and nephropathy, along with correlations with disease activity (SLEDAI), activation of some intracellular signaling proteins, and levels of serum inflammatory and oxidative markers were demonstrated. In vitro studies demonstrated the specific modulation of several genes/miRNAs by anti-dsDNA, along with the increase of prothrombotic and proinflammatory mediators, the induction of apoptosis and the phosphorylation of intracellular proteins participating in renal and CVD-related signaling pathways. Besides, treatment of HD-monocytes with SLE patients’ serum after Fluvastatin supplementation prevented the proinflammatory altered gene/miRNA profiles induced by serum from those patients before treatment. Conclusion: 1. Gene and microRNA expression profiles allowed the identification of relevant genes and pathways altered in monocytes of SLE patients, associated with the pathogenesis of the disease, and modulated by anti-dsDNA. 2. Specific microRNA-miRNA regulatory networks control the biological processes and factors related to the CV pathology in SLE, which are prevented by Fluvastatin treatment. Acknowledgement: Funded by ISCIII, PI15/01333, PI18/00837 and RIER RD16/0012/0015 co-funded with FEDER. Disclosure of Interests: None declared

Background The Splicing, a process of removing introns of RNA to produce mature messenger RNA, is exerted by an intricate macromolecular complex, the spliceosome. The alternative splicing, key mechanism that provides transcription and protein diversity, is dysregulated in various genetic, metabolic and inflammatory diseases. Splicing variants could contribute to the development and severity of pathologies of crucial importance. Objectives The aim of this study was the identification and characterization of alterations in the splicing machinery of leukocyte subsets of Rheumatoid Arthritis (RA) patients, as well as their influence on the activity of the disease and its atherothrombotic profile. Methods Monocytes, lymphocytes and neutrophils from 74 RA patients and 29 healthy donors (HD) were purified, and 45 elements of the splicing machinery were evaluated using a microfluidic qPCR array (Fluidigm). In parallel, the clinical status of the patients and the carotid intima-media thickness (CIMT) were assessed, along with an extensive serological evaluation, including autoantibodies, oxidative stress markers, inflammatory molecules and prothrombotic mediators. Correlation and association studies and logistic models among those clinical and analytical parameters were performed. Finally, leukocytes from HD were incubated with purified ACPAs-IgG from RA patients and changes promoted in both, splicing machinery and leukocyte inflammatory profile, were assessed. Results A significantly altered expression of spliceosome components was found in all the leukocyte subsets: 21, 17 and 27 components were differentially expressed in monocytes, lymphocytes and neutrophils, respectively, in RA patient’s vs HD. The levels of those altered components were associated to ACPA positivity, high disease activity (DAS28>5.1), early atherosclerosis and radiological involvement. Correlations with inflammatory mediators, oxidative stress markers and NETosis were further identified. Interestingly, eight components, belonging to the major spliceosome (SNRNP70, SNRNP200 and U2AF2), minor spliceosome (RNU4ATAC), and splicing factors (RBM3, RBM17, KHDRS1 and SRSF10) were found simultaneously altered in the three leukocyte subtypes. Logistic regression models, built with levels of simultaneously altered spliceosome components, generated several signatures related to RA disease features that were able to: i) discriminate between RA patients and HD; ii) classify patients with high disease activity (DAS28>5.1); iii) recognize patients with radiological involvement; and iv) identify patients showing atheroma plaques. In vitro studies showed that the purified ACPAs of RA patients modulated the expression of these components. Conclusion 1) The splicing machinery is profoundly altered in leukocytes from RA patients and closely related to the activity of the disease, including inflammation, autoimmunity and articular involvement, as well as with the occurrence of early atherosclerosis. 2) Evaluation of the common altered components of the spliceosome in leukocytes subsets might be used to typify the disease, avoiding complementary ultrasound or radiological tests. Acknowledgement MINECO (FJCI-2016-30825) and ISCIII (PI18/00837 and RIER RD16/0012/0015) co-funded with FEDER. Disclosure of Interests None declared