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Abstract Number: 0553

IgG from Systemic Sclerosis Patients Induce a Profibrosing and Serotype-dependent Phenotype in Normal Dermal Fibroblast: A Multi-omics Study

Aurélien Chepy1, Solange Vivier2, Fabrice Bray3, Martin Figeac4, Jean-Pascal Meneboo4, Camille Ternynck5, Lucile Guilbert2, Manel Jendoubi2, Christian Rolando3, David Launay1, Sylvain Dubucquoi6, Guillemette Marot5 and Vincent Sobanski1, 1Univ. Lille, Inserm, CHU Lille, Service de Médecine Interne, U1286 - INFINITE - Institute for Translational Research in Inflammation, F-59000 Lille, France, Lille, France, 2Univ. Lille, U1286 – INFINITE – Institute for Translational Research in Inflammation, F59000 Lille, France, Lille, France, 3Univ. Lille, CNRS, USR 3290 - MSAP - Miniaturisation pour la Synthèse, l'Analyse et la Protéomique, F-59000 Lille, France, Lille, France, 4Université de Lille, Functional Genomics Platform, Lille, France., Lille, France, 5ULR 2694 – METRICS : évaluation des technologies de santé et des pratiques médicales, université de Lille, CHU de Lille, 59000 Lille, France, Lille, France, 6Univ. Lille, U1286 – INFINITE – Institute for Translational Research in Inflammation, F59000 Lille, France, Lille

Meeting: ACR Convergence 2021

Keywords: Autoantibody(ies), Fibroblasts, Dermal, Genomics and Proteomics, Scleroderma, Systemic

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Session Information

Date: Sunday, November 7, 2021

Title: Systemic Sclerosis & Related Disorders – Basic Science Poster (0541–0559)

Session Type: Poster Session B

Session Time: 8:30AM-10:30AM

Background/Purpose: Antinuclear antibodies are frequent in systemic sclerosis (SSc). While recognized as potent biomarkers, their pathogenic role is much more debated. This study explored the effect of purified IgG from SSc patients on the phenotype and function of healthy dermal fibroblast (FB) using an innovative and unbiased multi-omics approach.

Methods: Normal dermal FB were cultured in the presence of purified IgG from patients with diffuse cutaneous SSc (dcSSc), limited cutaneous SSc or healthy controls (HC). FB proteome and transcriptome were explored using mass spectrometry coupled with liquid chromatography (LC-MS/MS) and microarray assays, respectively.

Results: Proteomic analysis identified 3310 differentially expressed proteins (DEP). SSc sera and purified IgG induced singular modifications FB protein profiles. These FB proteome changes were dependent of SSc serotype, especially pronounced with purified IgG from anti-topoisomerase I antibodies (ATA) positive patients (Figure 1). The enriched Gene Ontology (GO) terms upregulated in IgG purified SSc were involved in macroautophagy, positive regulation of protein transport and cell adhesion molecule transport. Transcriptomic analysis distinguished 764 differentially expressed genes and confirmed that IgG from dcSSc can induce profibrotic changes in genes profiles of FB. IgG from ATA positive patients induced singular changes: 629 DEP were identified in dcSSc ATA+ purified IgG and GO terms analysis showed enrichment in focal adhesion, cadherin binding, cytosolic part or lytic vacuole. Filamin A was the most strongly overexpressed protein in the presence of purified IgG compared to HC (Figure 2).

Conclusion: We identified that purified IgG from SSc can modify the phenotype of normal dermal FB. This effect seems dependent of the serotype. Purified IgG from dcSSc exhibited profibrotic properties with a singular profile of proteins expression and mRNA in patients with ATA.

Figure 1 legend: 1.A: heatmap representing 3310 differentially expressed proteins in proteomic analysis by Liquid chromatography–mass spectrometry (LC-MS/MS). Supervised clustering identified 5 different cluster of protein expression profile. 803, 894, 673, 626, 314 proteins were respectively found in cluster1, 2, 3, 4 and 5. 1.B: PCA of differentially expressed proteins. PCA highlights proteins expression by the FB according to SSc serotype. [ATA+] seemed to induce a singular protein expression profile compared to others SSc and SSc. PCA: principal component analysis. [HC] [Sera]: sera from healthy control; [HC] [IgG]: IgG from healthy control; [SSc] [Sera] [ATA+]: Sera from diffuse systemic sclerosis anti-Topoisomerase-I positive antibodies; [SSc] [IgG] [ATA+]: IgG from diffuse systemic sclerosis anti-Topoisomerase-I positive antibodies; [SSc] [Sera] [ATA-]: Sera from diffuse systemic sclerosis anti-Topoisomerase-I negative antibodies; [SSc] [IgG] [ATA-]: IgG from diffuse systemic sclerosis anti-Topoisomerase-I negative antibodies; [SSc] [Sera] [ACA+]: Sera from limited systemic sclerosis anti-centromere positive antibodies; [SSc] [IgG] [ACA+]: IgG from limited systemic sclerosis anti-centromere positive antibodies; [TGF-B]: fibroblast stimulated in the presence of TGF-β; [NS]: non stimulated fibroblast.

Figure 2 legend: A: heatmap (transcriptomic analysis) represented genes dysregulated in at least one contrast with FC>1.5 in the DNA probes dysregulated in proteomic analysis; B: Volcanoplot (proteomic analysis) represented the comparison [SSc] [IgG] [ATA+] vs [HC] [IgG]; proteins with adjust p-value <0.05 and Log Fold Change >1.4 or <_1.4 were considered; C: Enriched clusters GO terms in the comparison [SSc] [IgG] [ATA+] vs [HC] [IgG] according to upregulated proteins, analysis done with Metascape; [HC] [IgG]: IgG from healthy control; [SSc] [IgG] [ATA+]: IgG from diffuse systemic sclerosis anti-Topoisomerase-I positive antibodies; [SSc] [IgG] [ATA-]: IgG from diffuse systemic sclerosis anti-Topoisomerase-I negative antibodies; [SSc] [IgG]: IgG from systemic sclerosis patients; [NS]: non stimulated fibroblast.


Disclosures: A. Chepy, CSL Behring, 5; S. Vivier, None; F. Bray, None; M. Figeac, None; J. Meneboo, None; C. Ternynck, None; L. Guilbert, None; M. Jendoubi, None; C. Rolando, None; D. Launay, None; S. Dubucquoi, None; G. Marot, None; V. Sobanski, None.

To cite this abstract in AMA style:

Chepy A, Vivier S, Bray F, Figeac M, Meneboo J, Ternynck C, Guilbert L, Jendoubi M, Rolando C, Launay D, Dubucquoi S, Marot G, Sobanski V. IgG from Systemic Sclerosis Patients Induce a Profibrosing and Serotype-dependent Phenotype in Normal Dermal Fibroblast: A Multi-omics Study [abstract]. Arthritis Rheumatol. 2021; 73 (suppl 9). https://acrabstracts.org/abstract/igg-from-systemic-sclerosis-patients-induce-a-profibrosing-and-serotype-dependent-phenotype-in-normal-dermal-fibroblast-a-multi-omics-study/. Accessed .
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