Proteomic Analysis of Human Fibroblasts in Systemic Sclerosis Reinforces the Role of Transforming Growth Factor-ß and Points Toward Epidermal Growth Factor Receptor / Phosphatidylinositol 3 Kinase Pathway Inhibition

Benjamin Chaigne¹, Guilhem Clary², Morgane Le Gall³, Nicolas Dumoitier⁴, Sebastien Lofek⁵, Philippe Chafey², Pia Moinzadeh⁶, Thomas Krieg⁷, Christopher Denton⁸ and Luc Mouthon⁹, ¹Service de Médecine Interne, Centre de Référence Maladies Systémiques Autoimmunes Rares d’Ile de France, hôpital Cochin, DHU Authors, Assistance Publique-Hôpitaux de Paris, Paris, France, ²INSERM U1016, Institut Cochin,, Paris, France, ³INSERM U1016 Institut Cochin, Paris, France, ⁴INSERM U1016, Institut Cochin, Equipe Neutrophiles et Vascularites, Paris, France, ⁵INSERM U1016, paris, France, ⁶Department of Rheumatology, UCL Division of Medicine, London, United Kingdom, ⁷Universität zu Köln, Köln, Germany, ⁸Department of Rheumatology, University College London, Royal Free Hospital, London, United Kingdom, ⁹Service de Médecine Interne, Hôpital Cochin, Centre de référence national pour les maladies systémiques autoimmunes rares d’Ile de France, DHU Authors, Assistance Publique-Hôpitaux de Paris (AP-HP), Paris, France ;Université Paris Descartes Sorbonne Paris, Paris, France

Meeting: 2017 ACR/ARHP Annual Meeting

Date of first publication: September 18, 2017

Keywords: Fibroblasts, proteomics and systemic sclerosis

Session Information

Date: Sunday, November 5, 2017

Title: Systemic Sclerosis, Fibrosing Syndromes and Raynaud's – Pathogenesis, Animal Models and Genetics Poster I

Session Type: ACR Poster Session A

Session Time: 9:00AM-11:00AM

Background/Purpose:

Systemic sclerosis (SSc) is a rare autoimmune connective tissue disorder characterized by autoimmunity, vasculopathy and fibrosis. Fibrosis is due to an exaggerated activation of fibroblasts by the transforming growth factor-ß (TGF-ß). In this study we investigated the proteomic response of skin fibroblasts to TGF-ß in patients with SSc.

Methods:

Skin fibroblasts from 4 patients with SSc and 3 healthy controls (HC) were cultured in the presence or in the absence of TGF-ß. Two-dimensional gel electrophoresis and mass spectrometry (MS) were used to identify proteins differentially expressed between groups. Ingenuity Pathway analysis (IPA) and Panther softwares were used to analyse identified proteins. Finally real-time cell analyser (RTCA) was used to assess fibroblast proliferation and viability in order to validate proteins of interest.

Results:

We identified 687 protein spots differentially expressed between groups. 297 protein spots were analysed by MS. Principal component analysis revealed significant differences between fibroblasts of patients with SSc and HC when cultured in the presence or in the absence of TGF-ß. IPA revealed specific process networks such as actin cytoskeleton signalling, integrin signalling, and remodelling of epithelial adherens junctions. Panther revealed predominant biological processes such as cellular process, metabolic process and cellular component organization. TGF-ß enhanced the synthesis of fibroblasts’ proteins involved in actin cytoskeleton signalling and integrin signalling. Using IPA and RTCA we identified and validated the involvement of epidermal growth factor receptor (EGFR) and phosphatidylinositol 3 kinase (Pi3K) in the proliferation and the viability of fibroblasts from patients with SSc.

Conclusion:

In conclusion, we confirmed that the proteome profile of fibroblasts differs between patients with SSc and HC and demonstrated that fibroblasts enhance their proteomic phenotype upon stimulation with TGF-ß. We finally highlighted that EGFR and Pi3K are proteins of interest in fibroblasts from patients with SSc and should be considered as potential targets to inhibit fibrosis in SSc.

Disclosure: B. Chaigne, None; G. Clary, None; M. Le Gall, None; N. Dumoitier, None; S. Lofek, None; P. Chafey, None; P. Moinzadeh, None; T. Krieg, None; C. Denton, Actelion, Pfizer, GlaxoSmithKline, Bayer, Sanofi-Aventis, Boehringer Ingelheim, Genentech-Roche, CSL Behring, Biogen, 5,Actelion, GlaxoSmithKline, Bayer, Genentech-Roche, CSL Behring, 2; L. Mouthon, None.

To cite this abstract in AMA style:

Chaigne B, Clary G, Le Gall M, Dumoitier N, Lofek S, Chafey P, Moinzadeh P, Krieg T, Denton C, Mouthon L. Proteomic Analysis of Human Fibroblasts in Systemic Sclerosis Reinforces the Role of Transforming Growth Factor-ß and Points Toward Epidermal Growth Factor Receptor / Phosphatidylinositol 3 Kinase Pathway Inhibition [abstract]. Arthritis Rheumatol. 2017; 69 (suppl 10). https://acrabstracts.org/abstract/proteomic-analysis-of-human-fibroblasts-in-systemic-sclerosis-reinforces-the-role-of-transforming-growth-factor-s-and-points-toward-epidermal-growth-factor-receptor-phosphatidylinositol-3-kinase-pat/. Accessed .

« Back to 2017 ACR/ARHP Annual Meeting

ACR Meeting Abstracts - https://acrabstracts.org/abstract/proteomic-analysis-of-human-fibroblasts-in-systemic-sclerosis-reinforces-the-role-of-transforming-growth-factor-s-and-points-toward-epidermal-growth-factor-receptor-phosphatidylinositol-3-kinase-pat/