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SAT0319 Multiparametric detection of autoantibodies to investigate relationships between serological and clinical subsets of systemic sclerosis patients
  1. P Budde1,
  2. H-D Zucht1,
  3. DS Wirtz1,
  4. K Marquart1,
  5. M Schneider2,
  6. P Schulz-Knappe1,
  7. N Hunzelmann3
  1. 1Protagen Ag, Dortmund
  2. 2Rheumatology & Hiller-Forschungszentrum, Heinrich-Heine-University, Düsseldorf
  3. 3Dermatology, University of Cologne, Cologne, Germany

Abstract

Background Systemic sclerosis (SSc) is a largely heterogeneous autoimmune disease, with patients exhibiting an extensive range of clinical presentations and various disease course. The most widely used classification divides SSc into two major subsets diffuse cutaneous (dcSSc) and limited (lcSSc) SSc by the extent and severity of skin fibrosis. However, not all patients fit into these subsets. This has created great interest to examine disease heterogeneity at the molecular level to uncover unrecognized SSc subtypes that may differ with regard to clinical manifestations, prognosis or therapy response.

Objectives In large-scale “omics”-type autoantibody (AAB) profiling studies we have recently identified novel SSc-associated autoantigens. Here, we describe the development of a 20 marker multiplexed AAB assay to facilitate the discovery and validation of AAB-based patient subgroups.

Methods A Luminex bead-based AAB assay was designed by combining 8 connective tissue disease (anti-centromere, anti-Scl70, U1-snRNP, SSB, Ro52, Ro60, SmB, anti-ribosomal P) antigens with 12 novel antigens (including BICD2, JMJD3/KDM6B, and PPP1R2). Novel AAB targets were previously detected in SSc patients with a p-value <0.05 (Mann-Whitney-U-test) and frequency>15%. AAB reactivity was analysed in 92 SSc patients (dcSSc: n=32, lcSSc: n=50, SSc overlap: n=9). The mean modified Rodnan skin score (MRSS), mean disease duration (month), and mean age (years) of the SSc cohort was 10.51, 162.5 and 56.94, respectively. To analyze the individual-level patient similarity of AAB reactivity, the total number of AABs reactive in each patient was calculated and referenced to the number of all available antigens in percent. Patient's demographics and clinical data were dichotomized into patients with mRSS higher or lower than the mean value. Hierarchical cluster analysis was performed to investigate the relationship between AAB patient signatures and clinical and demographic features.

Results Based on their AAB reactivity pattern, the SSc sample cohort can be decomposed into four apparent clusters and additional fine-level clusters. LcSSc patients were spread over three clusters, each with clearly distinct AAB profile. Compared to dcSSc patients, lcSSc patients were more heterogeneous in their AAB profile. The percentage of lcSSc patients in clusters 1–3 was 70%, 90% and 53%, respectively. Patients in cluster 2 had an extended AAB repertoire that is anti-centromere, KDM6B, SSA, BICD2 and PPP1R2. 63% of all patients in cluster 4 were dcSSc and anti-Scl70 positive, who were most afflicted of the disease. AAB signatures of patients were mapped against dichotomized demographic and clinical features. Moving from cluster 1 to 4 the number of patients with shorter disease duration, lower age, higher mRSS and higher frequency of lung involvement increased.

Conclusions The multiplexed analysis of AABs in SSc enables defining an AAB reactivity score and patient clusters. This might support to subclassify SSc beyond lcSSc and dSSc.

Disclosure of Interest P. Budde Employee of: Protagen AG, H.-D. Zucht Employee of: Protagen AG, D. Wirtz Employee of: Protagen AG, K. Marquart Employee of: Protagen AG, M. Schneider Consultant for: Protagen AG, P. Schulz-Knappe Employee of: Protagen AG, N. Hunzelmann: None declared

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