Background/Purpose: This study aimed to 1) Analyze the circulating inflammatory profile of Rheumatoid Arthritis (RA) patients, in order to recognize distinctive clinical phenotypes associated with CV risk; 2) Evaluate the modulatory effects of TNF inhibitors (TNFi), JAK-STAT inhibitors (JAKInibs) and IL-6R inhibitors (IL6Ri); 3) Characterize underlying molecular mechanisms involved in endothelial dysfunction.

Methods: Two hundred and eight RA patients and 45 healthy donors (HD) were recruited. Serum inflammatory profile was assessed by analyzing 27 cytokines/chemokines -Luminex assays-, and biomolecules related to NETosis and oxidative stress -by base-plate kits-. Parallel extensive clinical analyses were performed. TNFi, JAKinibs and IL-6Ri effects were evaluated, respectively, in 45, 20 and 17 RA patients after 6 months. Lastly, mechanistic in vitro studies were developed in cultured endothelial cells (ECs) and changes in protein expression were evaluated in cell lysates by proximity extension assay technology, analyzing a panel of 88 proteins related to CV disease.

Results: Unsupervised-clustering identified 3 clusters representing specific molecular profiles. Clinically, even in the presence of similar disease score (DAS28) and positivity for autoantibodies, cluster 1 (C1) identified RA-patients expressing high inflammatory mediators’ levels, the highest CV-risk score, and a preponderance of atheroma plaques. Conversely, RA-patients conforming C3 showed the lowest inflammatory profile and the lowest CV-risk score. Lastly, C2 characterized an intermediate phenotype. Comparative analyses with a cohort of 98 RA patients presenting previous CV events, demonstrated that their inflammatory profile mimicked that found in C1, supporting the association of this altered shape with the CV status. In vivo, both biological and targeted-synthetic DMARDs’ therapy reduced DAS28-score and re-established normal levels of several altered biomolecules, reflecting a key role in the CV-risk control.

In vitro, RA patients’ serum pool from cluster 1 promoted in cultured ECs increased expression of several CV-related proteins, further prevented-albeit in an specific way- by the pre-incubation with TNFi (Etanercept), JAKinibs (Baricitinib) and anti-IL6Ri (Tocilizumab).

Conclusion:

1. The systemic inflammatory profile of RA identified patients’ subgroups with enhanced CV-risk, not associated with their disease activity status.

2. Both biological and targeted-synthetic DMARDs re-established normal levels of circulating inflammatory biomolecules, reducing the CV-risk in RA.

3. In vitro studies revealed that RA-serum inflammatory mediators directly induced endothelial damage which might be prevented by effect of both, biological and targeted-synthetic DMARDs’ therapy.

Thus, the analysis of the RA patients circulating molecular profile might contribute to improving the personalized clinical management of these patients and their CV risk.

Supported by ISCIII (PI21/005991 and RICOR-RD21/0002/0033) co-financed by FEDER, Fundacion Andaluza de Reumatología (FAR) and Consejería de Conocimiento, Investigación y Universidad de la Junta de Andalucía (P20_01367).

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ACR Meeting Abstracts - https://acrabstracts.org/abstract/unsupervised-clustering-analysis-of-circulating-molecular-profiles-in-rheumatoid-arthritis-patients-defines-precise-cv-risk-effects-of-tnfi-il6ri-and-jakinibs/