Effects of Biological Therapies on Molecular Features of Rheumatoid Arthritis

Rheumatoid arthritis (RA) is an autoimmune and chronic inflammatory disease primarily affecting the joints, and closely related to specific autoantibodies that mostly target modified self-epitopes. Relevant findings in the field of RA pathogenesis have been described. In particular, new insights come from studies on synovial fibroblasts and cells belonging to the innate and adaptive immune system, which documented the aberrant production of inflammatory mediators, oxidative stress and NETosis, along with relevant alterations of the genome and on the regulatory epigenetic mechanisms. In recent years, the advances in the understanding of RA pathogenesis by identifying key cells and cytokines allowed the development of new targeted disease-modifying antirheumatic drugs (DMARDs). These drugs considerably improved treatment outcomes for the majority of patients. Moreover, numerous studies demonstrated that the pharmacological therapy with biologic DMARDs (bDMARDs) promotes, in parallel to their clinical efficacy, significant improvement in all these altered molecular mechanisms. Thus, continuous updating of the knowledge of molecular processes associated with the pathogenesis of RA, and on the specific effects of bDMARDs in the correction of their dysregulation, are essential in the early and correct approach to the treatment of this complex autoimmune disorder. The present review details basic mechanisms related to the physiopathology of RA, along with the core mechanisms of response to bDMARDs

Therapeutic Potential and Immunomodulatory Role of Coenzyme Q10 and Its Analogues in Systemic Autoimmune Diseases

Coenzyme Q10 (CoQ10) is a mitochondrial electron carrier and a powerful lipophilic antioxidant located in membranes and plasma lipoproteins. CoQ10 is endogenously synthesized and obtained from the diet, which has raised interest in its therapeutic potential against pathologies related to mitochondrial dysfunction and enhanced oxidative stress. Novel formulations of solubilized CoQ10 and the stabilization of reduced CoQ10 (ubiquinol) have improved its bioavailability and efficacy. Synthetic analogues with increased solubility, such as idebenone, or accumulated selectively in mitochondria, such as MitoQ, have also demonstrated promising properties. CoQ10 has shown beneficial effects in autoimmune diseases. Leukocytes from antiphospholipid syndrome (APS) patients exhibit an oxidative perturbation closely related to the prothrombotic status. In vivo ubiquinol supplementation in APS modulated the overexpression of inflammatory and thrombotic risk-markers. Mitochondrial abnormalities also contribute to immune dysregulation and organ damage in systemic lupus erythematosus (SLE). Idebenone and MitoQ improved clinical and immunological features of lupus-like disease in mice. Clinical trials and experimental models have further demonstrated a therapeutic role for CoQ10 in Rheumatoid Arthritis, multiple sclerosis and type 1 diabetes. This review summarizes the effects of CoQ10 and its analogs in modulating processes involved in autoimmune disorders, highlighting the potential of these therapeutic approaches for patients with immune-mediated diseases

The clinical and molecular cardiometabolic fingerprint of an exploratory psoriatic arthritis cohort is associated with the disease activity and differentially modulated by methotrexate and apremilast

Objectives: (1) To evaluate clinical and molecular cardiovascular disease (CVD) signs and their relationship with psoriatic arthritis (PsA) features and (2) to identify a clinical patient profile susceptible to benefit from methotrexate (MTX) and/or apremilast regarding CVD risk. Methods: This cross-sectional study included 100 patients with PsA and 100 age-matched healthy donors. In addition, an exploratory cohort of 45 biologically naïve patients treated for 6 months with apremilast, MTX or combined therapy according to routine clinical practice was recruited. Extensive clinical and metabolic profiles were obtained. Ninety-nine surrogate CVD-related molecules were analysed in plasma and peripheral blood mononuclear cells (PBMCs). Hard cluster analysis was performed to identify the clinical and molecular phenotypes. Mechanistic studies were performed on adipocytes. Results: Cardiometabolic comorbidities were associated with disease activity and long-term inflammatory status. Thirty-five CVD-related proteins were altered in the plasma and PBMCs of PsA patients and were associated with the key clinical features of the disease. Plasma levels of some of the CVD-related molecules might distinguish insulin-resistant patients (MMP-3, CD163, FABP-4), high disease activity (GAL-3 and FABP-4) and poor therapy outcomes (CD-163, LTBR and CNTN-1). Hard cluster analysis identified two phenotypes of patients according to the rates of cardiometabolic comorbidities with distinctive clinical and molecular responses to each treatment. Conclusions: (1) Novel CVD-related proteins associated with clinical features could be emerging therapeutic targets in the context of PsA and (2) the pleiotropic action of apremilast could make it an excellent choice for the management of PsA patients with high CVD risk, targeting metabolic alterations and CVD-related molecules

Complement component 3 as biomarker of disease activity and cardiometabolic risk factor in rheumatoid arthritis and spondyloarthritis.

Funder: Europeo de Desarrollo Regional de la Unión EuropeaFunder: Rheumatic Diseases NetworkOBJECTIVE: To analyze the relationship between complement component 3 (C3) and the prevalence of cardiometabolic risk factors and disease activity in the rheumatic diseases having the highest rates of cardiovascular morbidity and mortality: rheumatoid arthritis (RA), psoriatic arthritis (PsA) and axial spondyloarthritis (axSpA). METHODS: This is a cross-sectional study including 200 RA, 80 PsA, 150 axSpA patients and 100 healthy donors. The prevalence of cardiometabolic risk factors [obesity, insulin resistance, type 2 diabetes mellitus, hyperlipidemia, apolipoprotein B/apolipoprotein A (apoB/apoA) and atherogenic risks and hypertension] was analyzed. Serum complement C3 levels, inflammatory markers and disease activity were evaluated. Cluster analysis was performed to identify different phenotypes. Receiver operating characteristic (ROC) curve analysis to assess the accuracy of complement C3 as biomarker of insulin resistance and disease activity was carried out. RESULTS: Levels of complement C3, significantly elevated in RA, axSpA and PsA patients, were associated with the prevalence of cardiometabolic risk factors. Hard clustering analysis identified two distinctive phenotypes of patients depending on the complement C3 levels and insulin sensitivity state. Patients from cluster 1, characterized by high levels of complement C3 displayed increased prevalence of cardiometabolic risk factors and high disease activity. ROC curve analysis showed that non-obesity related complement C3 levels allowed to identify insulin resistant patients. CONCLUSIONS: Complement C3 is associated with the concomitant increased prevalence of cardiometabolic risk factors in rheumatoid arthritis and spondyloarthritis. Thus, complement C3 should be considered a useful marker of insulin resistance and disease activity in these rheumatic disorders

Advances in Understanding the Role of NRF2 in Liver Pathophysiology and Its Relationship with Hepatic-Specific Cyclooxygenase-2 Expression

Oxidative stress and inflammation play an important role in the pathophysiological changes of liver diseases. Nuclear factor erythroid 2-related factor 2 (NRF2) is a transcription factor that positively regulates the basal and inducible expression of a large battery of cytoprotective genes, thus playing a key role in protecting against oxidative damage. Cyclooxygenase-2 (COX-2) is a key enzyme in prostaglandin biosynthesis. Its expression has always been associated with the induction of inflammation, but we have shown that, in addition to possessing other benefits, the constitutive expression of COX-2 in hepatocytes is beneficial in reducing inflammation and oxidative stress in multiple liver diseases. In this review, we summarized the role of NRF2 as a main agent in the resolution of oxidative stress, the crucial role of NRF2 signaling pathways during the development of chronic liver diseases, and, finally we related its action to that of COX-2, where it appears to operate as its partner in providing a hepatoprotective effect

Liver spheroids: a robust human in vitro system for testing the therapeutic options of cyclooxygenase 2 in NAFLD/NASH

Trabajo presentado en el FASEB Science Research Conference: The Liver Biology Conference: Fundamental Mechanisms and Translational Applications,[Background]: NAFLD is the most prevalent chronic liver disease in the world and is considered the hepatic manifestation of metabolic syndrome. It begins with an accumulation of fat in the liver, which can lead to cellular damage and inflammation (NASH) and, if unaddressed, progresses to liver fibrosis, where parenchymal tissue is replaced by extracellular matrix. Cyclooxygenase (COX) is a key regulatory step in the biosynthesis of prostanoids. The COX-2 isoform is expressed and induced by different stimuli in various tissues and cell types; however, in liver, COX-2 expression is restricted to those situations in which proliferation or dedifferentiation occur (1). Our previous results revealed that COX-2 expression in hepatocytes protects against hyperglycemia-induced liver damage and against peripheral insulin resistance and adiposity in mice subjected to a high-fat diet (2). Furthermore, COX-2 expression in hepatocytes protects against experimental nonalcoholic steatohepatitis and fibrosis in mice (3). [Aims]: Since our previous results are performed in mice, we wanted to bridge from animal models to a more translational research, close to human. Therefore, we wanted to establish an in vitro model based on 3D human cell culture to evaluate the role of COX-2, specifically the role of prostaglandins, in a human model of NAFLD/NASH. [Methods]: To analyze the role of COX-2 in steatosis and fibrosis, we established a 3D spheroid culture composed of HepG2 and LX-2 cells. These spheroids were treated with palmitic acid and TGF¿ to induce NAFLD/NASH and fibrosis, respectively. They were then treated with PGE2 to evaluate their effect in this model. [Results]: In summary, our results demonstrate that PGE2 treatment reduces lipid levels after fatty acid exposure. Moreover, it reduces fatty acid-induced cell damage and insulin resistance. Furthermore, PGE2 slows fibrotic progression by decreasing stellate cells activation after TGF¿ stimulation. Conclusions: These preliminary results indicate that a therapeutic strategy for the treatment of NAFLD/NASH may be COX-2-derived prostaglandin therapy.This work was supported by SAF2016-75004-R and PID2019-108977RB-I00 (MINECO, Spain) and CIBERehd (ISCIII, Spain). M F-A. and M L-T are recipient of FPI fellowship from MINECO (BES-2017-081928 and PRE2020-094885, respectively). We thank FEDER for financial support

COX-2 como agente terapeútico en la fisiopatología de hígado graso no alcohólico (G01 49 P 041)

Trabajo presentado en el 44º Congreso de la SEBBM, celebrado en Málaga (España) del 06 al 09 de septiembre de 2022.(G01 49 P 041) [Introducción]: El hígado graso no alcohólico (EHGNA) es actualmente la enfermedad hepática crónica más prevalente. El espectro de EHGNA abarca desde esteatosis, a esteatohepatitis, cirrosis, y finalmente hepatocarcinoma. La ciclooxigenasa 2 (COX-2), enzima de creciente interés por su papel dual en hígado, se ha detectado en EHGNA. Sin embargo, su papel en dicha patología es aún controvertido. Nuestros estudios previos han demostrado que la expresión constitutiva de COX-2 en los hepatocitos protege frente al daño hepático inducido por hiperglucemia, resistencia a insulina y la acumulación lipídica en ratones con dieta rica en grasa. [Objetivo]: Evaluar el papel de COX-2 como potencial diana terapéutica en la patología de EHGNA a través de la generación de un modelo murino transgénico inducible (ihCOX-2-Tg), en el que se provoque (antes de la expresión de la COX-2) esteatosis y fibrosis a través de la alimentación con una dieta rica en grasa y la administración de tetracloruro de carbono (CCL4), respectivamente. Materiales: La esteatosis y fibrosis moderada propias de la patología de EHGNA se lograron a través de una dieta rica en grasa (L-Amino Acid Diet With 60 kcal% Fat With 0.1% Methionine and No Added Choline, A06071302i Research Diet) durante 6 semanas. La fibrosis severa se provocó a través de la administración intraperitoneal (IP) de CCL4 durante 9 semanas. La expresión de COX-2 se indujo en el modelo ihCOX-2-Tg mediante la administración intravenosa (IV) de 1011 copias de adenovirus asociados serotipo 8 (AAV8) CRE a la semana o a las 3 semanas del inicio de la dieta o tras 4 semanas de tratamiento con CCL4. El daño hepático se evaluó mediante parámetros bioquímicos e histológicos, así como ensayos de biología molecular. [Resultados]: Nuestro modelo ihCOX-2-Tg expresó COX-2 hepática tras la inoculación de AAV8-CRE. Además, los adenovirus asociados empleados presentaron tropismo hepático. Nuestros resultados preliminares indicaron que los ratones inoculados con AAV8-CRE presentaron un menor porcentaje de fibrosis hepática, que se acompañó de una disminución en el tamaño de las gotas lipídicas y de marcadores de fibrosis y acumulación lipídica, así como de un aumento de las PGs hepáticas respecto a aquellos ratones inoculados con el control AAV8-GFP. Conclusiones: Nuestros resultados preliminares evidencian el prometedor papel de COX-2 como diana terapéutica en EHGNA a través de la amortiguación de la esteatosis y fibrosis hepática en nuestro modelo murino

Liver spheroids: a robust human in vitro system for testing the therapeutic options of cyclooxygenase 2 in NAFLD/NASH

Trabajo presentado en las XV Jornadas CIBER Enfermedades Hepáticas y Digestivas: Jornades científiques del CIBERehd, celebradas en modalidad virtual y de forma presencial en Barcelona (España) del 08 al 09 de noviembre de 2021.Cyclooxygenase (COX) is a key enzyme in the biosynthesis of prostanoids. Prostaglandins are involved in multiple homeostatic processes, as well as playing an important role in the onset of inflammation. COX-2 is an isoform that is expressed and induced by different stimuli in various tissues and cell types; however, in liver, COX-2 expression is restricted to those situations where proliferation and dedifferentiation occur (1). Our previous results have shown that COX-2 expression in hepatocytes protects against hyperglycemia-induced liver damage, peripheral insulin resistance and adiposity in mice fed a high-fat diet (2), and also protects against experimental non-alcoholic steatohepatitis and fibrosis (3).This work was supported by SAF2016-75004-R and PID2019-108977RB-I00 (MINECO, Spain) and CIBERehd (ISCIII, Spain). M F-A. and M L-T are recipient of FPI fellowship from MINECO (BES-2017-081928 and PRE2020-094885, respectively). We thank FEDER for financial support

Early restoration of immune and vascular phenotypes in systemic lupus erythematosus and rheumatoid arthritis patients after B cell depletion.

This translational multi-centre study explored early changes in serologic variables following B lymphocyte depletion by rituximab (RTX) treatment in systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) patients and investigated in vitro effects on the activity of other immune cells and the vascular endothelium. Eighty-five SLE patients, seventy-five RA patients and ninety healthy donors were enrolled. Two additional cohorts of selected SLE and RA patients were treated with RTX for 3 months. Changes in circulating levels of inflammatory mediators, oxidative stress markers and NETosis-derived bioproducts were evaluated. Serum miRNomes were identified by next-generation sequencing, and RTX-induced changes were delineated. Mechanistic in vitro studies were performed to assess activity profiles. Altered inflammatory, oxidative and NETosis-derived biomolecules were found in SLE and RA patients, closely interconnected and associated to specific miRNA profiles. RTX treatment reduced SLE and RA patients' disease activity, linked to a prominent alteration in those biomolecules and the reversal of altered regulating miRNAs. In vitro studies showed inhibition of NETosis and decline of pro-inflammatory profiles of leucocytes and human umbilical vein endothelial cells (HUVECs) after B cell depletion. This study provides evidence supporting an early RTX-induced re-setting of the pro-inflammatory status in SLE and RA, involving a re-establishment of the homeostatic equilibrium in immune system and the vascular wall

Early restoration of immune and vascular phenotypes in systemic lupus erythematosus and rheumatoid arthritis patients after B cell depletion

This translational multi-centre study explored early changes in serologic variables following B lymphocyte depletion by rituximab (RTX) treatment in systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) patients and investigated in vitro effects on the activity of other immune cells and the vascular endothelium. Eighty-five SLE patients, seventy-five RA patients and ninety healthy donors were enrolled. Two additional cohorts of selected SLE and RA patients were treated with RTX for 3 months. Changes in circulating levels of inflammatory mediators, oxidative stress markers and NETosis-derived bioproducts were evaluated. Serum miRNomes were identified by next-generation sequencing, and RTX-induced changes were delineated. Mechanistic in vitro studies were performed to assess activity profiles. Altered inflammatory, oxidative and NETosis-derived biomolecules were found in SLE and RA patients, closely interconnected and associated to specific miRNA profiles. RTX treatment reduced SLE and RA patients' disease activity, linked to a prominent alteration in those biomolecules and the reversal of altered regulating miRNAs. In vitro studies showed inhibition of NETosis and decline of pro-inflammatory profiles of leucocytes and human umbilical vein endothelial cells (HUVECs) after B cell depletion. This study provides evidence supporting an early RTX-induced re-setting of the pro-inflammatory status in SLE and RA, involving a re-establishment of the homeostatic equilibrium in immune system and the vascular wall