Background/Purpose: 1. To identify and characterize specific immunological, inflammatory and epigenetic determinants involved in the enhanced cardiovascular (CV)-risk present in Rheumatoid Arthritis (RA) patients. 2. To evaluate the effects of biological drugs on the reestablishment of this altered profile.

Methods: Serum samples from 52 healthy donors (HD) and 188 selected RA patients -including 100 patients that had suffered previous CV events and 88 without previous CV events- were studied. Inflammatory and oxidative stress biomolecules and Netosis-derived products were quantified, and the CV-Risk Score was calculated following EULAR recommendations. Carotid intima-media thickness (CIMT) was evaluated as early atherosclerosis marker. miRNnomes were identified using next-generation sequencing miRNA assay. The in vivo effects of biologic drugs such as Infliximab (IFX), Tocilizumab (TCZ) and Rituximab (RTX) were evaluated before and after 6 months of therapy in parallel cohorts of 45, 25 and 27 patients, respectively.

Results: Circulating biomolecules related to inflammation -interleukins, chemokines, adhesion molecules-, Netosis -cell-free nucleosomes and elastase- and oxidative stress -lipoperoxides and 8-hydroxy-2′-deoxyguanosine (8-OHdG)- were found coordinately altered in the serum of RA patients. 104 circulating miRNAs were found altered in RA patients. Functional classification (Ingenuity Pathway analysis, IPA) recognized their potential involvement in inflammatory response, immunological and hematological diseases, and correlation analyses established the relationship of a number of them with the biomolecules found altered in RA patients. Non-supervised hard clustering analysis differentiated 3 clusters representing different CV-risk profile groups: a) RA patients with elevated CV-risk score ( >9), positive for autoantibodies (RF and ACPAs) and with a high incidence (53 %) of CV events; b) RA patients with medium-low CV-risk score (< 4) but positive for autoantibodies and also with a high incidence (47%) of CV events, thus suggesting a relevant role for autoimmunity in this increased CV-risk; and c) RA patients negative for autoantibodies with medium-low CV-risk score and without previous CV events. Those three phenotypes of RA patients further displayed distinctive and specific molecular shapes. In vivo treatments with IFX, TCZ and RTX reduced disease activity and induced the re-establishment of normal levels in these altered biomolecules in an inhibitor-dependent manner.

Conclusion:

1. Phenotype-based clustering combined with the analysis of molecular profiles might help to define precise CV-risk profiles in RA patients. 2. Specific mediators of autoimmunity, inflammation, oxidative damage and Netosis, along with the miRNAs modulating their expression, coordinately contribute to a higher CV-risk score in RA patients. 3. Biological drugs, most likely though distinctive molecular mechanisms, restore the normal levels of these altered biomolecules, reducing the CV risk in RA patients. Funded by PI-0285-2017, ISCIII, PI18/00837 and RIER RD16/0012/0015 co-funded with FEDER

ACR Meeting Abstracts - https://acrabstracts.org/abstract/phenotype-based-clustering-along-with-analysis-of-molecular-profile-might-help-to-define-precise-cv-risk-profiles-in-ra-patients/