Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Personalized cardiovascular risk assessment in Rheumatoid Arthritis patients using circulating molecular profiles and their modulation by TNFi, IL6Ri, and JAKinibs.

doi: 10.1016/j.biopha.2024.116357

Figure Lengend Snippet: Fig. 5. Serum from RA patients with high CVD risk induced the upregulation of CVD-related markers in endothelial cells (ECs) and monocytes, which was prevented by biological and targeted synthetic DMARDs.- (A and C) Heat map showing the changes promoted in CV- related proteins on endothelial cells (A) and monocytes (C) treated in vitro with serum from RA patients belonging to cluster 3 (showing the highest inflammatory profile) in comparison with serum from HD. Levels of inflammatory proteins are expressed as NPX (normalized protein expression, arbitrary units on a Log 2 scale), and have undergone a clustering analysis to aid interpretation. Proteins showing significant alterations (FDR-adjusted p value <0.05) are detached in bold. (B and D) Predicted and validated protein-protein interactions among proteins potentially modulated by RA-C3 serum, using the STRING platform. Protein networks showing the relationship between differen tially expressed proteins are displayed. Below are displayed tables containing functional enrichment analysis of biological processes by using the Gene ontology platform. (E and G) Changes in cardiovascular mediators’ levels in cell lysates of endothelial cells and monocytes cultured with HD or RA-C3 serum, either, in the presence or in the absence of TNFi, IL6Ri or JAKinibs. *p<0.05, **p<0.01, ***p<0.001. (F and H) Venn diagram showing the common and differential proteins modulated in endothelial cells and monocytes by TNFi, IL6Ri or JAKinibs.

Article Snippet: Neutrophils and monocytes purified from HD and primary human umbilical vein endothelial cells (HUVECs) were subjected to a 12-h treatment (neutrophils) or 24-h treatment (monocytes and HUVECs)at 37 ◦C. with the serum from HD, the serum of RA patients that had suffered previous CV events (RA-CVD), or the serum from patients belonging to the cluster 3 (RA-C3), either in the presence or absence of TNFi (Etanercept, - European Pharmacopoeia reference standard, Strasbourg, France- 10 ug/mL), IL6Ri (Sarilumab, -Selleckchem, Planegg, Germany- 10 ug/mL,) or JAKinibs (Baricitinib, -Tocris Bioscience, Bristol, UK- 10 ug/mL).

Techniques: In Vitro, Comparison, Expressing, Protein-Protein interactions, Functional Assay, Cell Culture

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Personalized cardiovascular risk assessment in Rheumatoid Arthritis patients using circulating molecular profiles and their modulation by TNFi, IL6Ri, and JAKinibs.

doi: 10.1016/j.biopha.2024.116357

Figure Lengend Snippet: Fig. 4. Changes in clinical and molecular profiles of rheumatoid arthritis patients treated with b-DMARDs or ts-DMARDs. (A) RA patients’ (n = 83) changes in clinical features at 6 months of TNFi, IL6Ri and JAKinibs therapies, including Disease Activity Score (DAS28), Clinical Disease Activity Index (CDAI), and Simple Disease Activity index (SDAI). At the initial timepoint (t0), each data point corresponds to an individual patient, with a subsequent connection established to the respective measurement obtained at the 6-month interval. (B) Heat map showing differential levels of circulating inflammatory proteins in plasma of patients with RA after 6 months of TNFi, IL6Ri and JAKinibs therapies (Δ T6-T0). Levels of inflammatory proteins are expressed as log 2,and have undergone a clustering analysis to aid interpretation. (C) RA patients’ changes in NETosis bioproducts (elastase and nucleosomes) and lipoperoxides after 6 months of TNFi, IL6Ri and JAKinibs therapies. * p ≤0.05, ** p ≤0.01, *** p ≤0.001, **** p ≤0.0001.

Article Snippet: Neutrophils and monocytes purified from HD and primary human umbilical vein endothelial cells (HUVECs) were subjected to a 12-h treatment (neutrophils) or 24-h treatment (monocytes and HUVECs)at 37 ◦C. with the serum from HD, the serum of RA patients that had suffered previous CV events (RA-CVD), or the serum from patients belonging to the cluster 3 (RA-C3), either in the presence or absence of TNFi (Etanercept, - European Pharmacopoeia reference standard, Strasbourg, France- 10 ug/mL), IL6Ri (Sarilumab, -Selleckchem, Planegg, Germany- 10 ug/mL,) or JAKinibs (Baricitinib, -Tocris Bioscience, Bristol, UK- 10 ug/mL).

Techniques: Activity Assay, Clinical Proteomics

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Personalized cardiovascular risk assessment in Rheumatoid Arthritis patients using circulating molecular profiles and their modulation by TNFi, IL6Ri, and JAKinibs.

doi: 10.1016/j.biopha.2024.116357

Figure Lengend Snippet: Fig. 6. Serum from RA patients with high CVD risk induced NETosis and neutrophils activation, which was prevented by b-DMARDs and ts-DMARDs.- (A- B) Bar graphs showing the changes promoted in gene expression levels of several activation markers on neutrophils treated in vitro with serum from RA patients belonging to cluster 3 (showing the highest inflammatory profile) either in the presence or in the absence of TNFi, IL6Ri or JAKinibs inhibitors. All experiments were compared with neutrophils treated with HD serum, set at 100% in each panel. (Significance at p<0.05, a, vs neutrophils treated with HD serum; b, vs neutrophils treated with RA-C3 serum). (C) Representative micrographs of neutrophil extracellular traps (NETs) from HD neutrophils treated in vitro with serum from RA patients belonging to cluster 3 (showing the highest inflammatory profile) either in the presence or in the absence of TNFi, IL6Ri or JAKinibs inhibitors. NETs were visualized by using a Nikon Eclipse-Ti-S fluorescence microscope 20x objective. Scale bar 10 micrometers. (D) Concentration of cell-free nucleosomes and cell-free elastase in supernatants from cell cultures of HD neutrophils performed as above described (Significance at p<0.05, a, vs neutrophils treated with HD serum; b, vs neutrophils treated with RA-C3 serum).

Article Snippet: Neutrophils and monocytes purified from HD and primary human umbilical vein endothelial cells (HUVECs) were subjected to a 12-h treatment (neutrophils) or 24-h treatment (monocytes and HUVECs)at 37 ◦C. with the serum from HD, the serum of RA patients that had suffered previous CV events (RA-CVD), or the serum from patients belonging to the cluster 3 (RA-C3), either in the presence or absence of TNFi (Etanercept, - European Pharmacopoeia reference standard, Strasbourg, France- 10 ug/mL), IL6Ri (Sarilumab, -Selleckchem, Planegg, Germany- 10 ug/mL,) or JAKinibs (Baricitinib, -Tocris Bioscience, Bristol, UK- 10 ug/mL).

Techniques: Activation Assay, Gene Expression, In Vitro, Fluorescence, Microscopy, Concentration Assay