THU0525 DEVELOPMENT OF A PREDICTIVE TOOL FOR RESPONSE TO ANTI-TNF-ALPHA THERAPY IN JIA USING GENE EXPRESSION PROFILES IN PERIPHERAL DERIVED MONONUCLEAR CELLS
Authors: Jedidja Baaij, Rianne Scholman, Rae Yeung, Trang Duong, Nico Wulffraat, Joost F. Swart, Sebastian Vastert, Sytze De Roock
Journal: Annals of the Rheumatic Diseases (2019)

<h3>Background:</h3> Heterogeneity in response to biological medication is a major challenge in the management of Juvenile Idiopathic Arthritis (JIA) (1). Biomarkers such as cytokines or whole blood RNA expression profiles that predict therapy efficacy prior to treatment pose a solution and could enable precision medicine. It was previously demonstrated that whole blood gene expression profiles can predict response to anti-TNF-α therapy in patients with JIA (2). These findings currently await validation in a multicenter prospective cohort study. Harmonization of data from different cohorts, however, is often hampered by site-dependent differences in biological sample collection and processing (3). For instance, the biological material that is retrospectively available from biobanks depends on what is stored exactly, e.g. whole blood versus peripheral blood mononuclear cells (PBMC). Furthermore, different blood collection tubes containing different additives can affect sample quality at an early stage and make interchangeability or comparability of data impossible. <h3>Objectives:</h3> We evaluated the comparability of gene expression profiles when whole blood from patients with JIA is collected in different RNA collection tubes and processed accordingly. Next, we will investigate the predictive capacity for therapy response of RNA expression profiles from frozen PBMC of 63 JIA patients. <h3>Methods:</h3> Peripheral blood from 11 children with non-systemic JIA with active disease was collected in PAXgene (PreAnalytiX) and Tempus (Applied Biosystems) tubes. All tubes were subsequently stored in the freezer at -80 °C. After thawing, RNA from the PAXgene tubes was extracted with use of the PAXgene Blood RNA Kit and RNA from the Tempus tubes was isolated by using the Tempus Preserved Blood RNA Purification Kit I. Gene expression of <i>CSNK1D</i>, <i>C1D</i>, <i>ASAP2</i>, <i>SRRD</i>, <i>PPP1R3B</i>, <i>HLA-DQA1</i>, <i>PDZK1IP1</i> and <i>MZB1</i> was determined with qPCR. For our next experiment, frozen PBMC derived from patients with non-systemic JIA who are included in our longitudinal Pharmachild registry biobank will be used. Of the 63 patients, 28 (44.4%) children were diagnosed with oligo articular JIA, 19 (30.2%) with poly articular JIA, 9 (14.3%) with enthesitis-related JIA, 4 (6.3%) with psoriatic JIA and 3 (4.8%) with undifferentiated JIA. The average age at sampling was 11.5 years ± 4.0 and 65% of the patients was female. These samples will be run on a 96-analyte NanoString panel and associated with clinical response at time points 3 and 6 months after start of TNF-α blockade. <h3>Results:</h3> Both RNA blood collection systems yielded high-quality RNA, with overall higher RNA concentrations for blood collected in Tempus tubes in comparison to PAXgene tubes. qPCR data showed that gene expression of all measured genes is affected by the method of blood collection and processing. However, the inter-individual variation was similar between both collection tubes, indicating that similar RNA profiles were observed. <h3>Conclusion:</h3> Gene expression profiles derived from whole blood collected in PAXgene or Tempus tubes are comparable, but are not interchangeable. <h3>References:</h3> [1] Swart JF, de Roock S, &amp; Prakken BJ. Understanding inflammation in juvenile idiopathic arthritis: How immune biomarkers guide clinical strategies in the systemic onset subtype. Eur J Immunol 2016;46(9):2068-2077. [2] Cui A, Quon G, Rosenberg AM et al. Gene Expression Deconvolution for Uncovering Molecular Signatures in Response to Therapy in Juvenile Idiopathic Arthritis. PloS one 2016;11(5):1-17. [3] Yeung RS, Albani S, Feldman BM et al. Enhancing translational research in paediatric rheumatology through standardization. Nature Reviews Rheumatology 2016;12(11):684. <h3>Disclosure of Interests:</h3> None declared