Role of Eosinophil in Fibrogenesis of Systemic Sclerosis

Study Overview
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Brief Summary
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Eosinophils are involved in tissue remodeling and fibrosis in many inflammatory diseases. Systemic sclerosis (SSc) is an autoimmune disease with fibrotic skin and lung complications. The profibrosing properties and data from the SSc literature suggest a possible role of the eosinophils in the process of fibrogenesis of SSc.

Detailed Description
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it will assess the activation state of blood eosinophils in SSc patients compared to healthy controls (ECP production in vitro, surface activation markers, whole transcriptome array, ..). it will also study skin eosinophils and their recruitment (extracellular eosinophil granules, eotaxins production in skin, ..)

Official Title
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Role of Blood and Tissue Eosinophils in the Fibrogenesis of Systemic Sclerosis

Conditions
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Systemic Sclerosis, Systemic Scleroderma

Intervention / Treatment
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* Diagnostic Test: Blood test
* Diagnostic Test: Skin biopsies


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: For SSc patients: Meeting the 2013 ACR-EULAR criteria for diffuse SSc (n = 20) and limited SSc (n = 20) Having signed the informed consent Eosinophils ≥ 0.1 G / L on the last blood test (<3 months) For healthy subjects: Without chronic pathology requiring long-term treatment Eosinophils between 0.1 and 0.4 G / L on recent blood test (<3 months) Exclusion Criteria for patients and controls: Patient with an infection, state of emergency or progressive neoplastic pathology Pregnant or lactating women History of atopy (allergic asthma, atopic eczema, allergic rhinitis, allergic conjunctivitis) Allergy to local anesthetics (for scleroderma patients only) Taking into account a minimum weight of 50 kg and in the absence of cardiorespiratory effects of scleroderma, only patients with at least 10 g of hemoglobin will be included

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 66 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Diffuse SSc<br>Recruitment of 20 patients with diffuse SSc | Diagnostic Test: Blood test<br>* Patients in each group will have a blood sample (9x7 mL) for eosinophils isolation, study of activation markers on whole blood and serum biomarkers testing<br>Diagnostic Test: Skin biopsies<br>* In patients who will accept, skin biopsies will be performed in damaged and apparently normal skin, excluding fingers, hands, feet and face (biopsies are facultative)<br>|
| Limited SSc<br>Recruitment of 20 patients with limited SSc | Diagnostic Test: Blood test<br>* Patients in each group will have a blood sample (9x7 mL) for eosinophils isolation, study of activation markers on whole blood and serum biomarkers testing<br>Diagnostic Test: Skin biopsies<br>* In patients who will accept, skin biopsies will be performed in damaged and apparently normal skin, excluding fingers, hands, feet and face (biopsies are facultative)<br>|
| Healthy subjects<br>Recruitment of 20 healthy subjects (control) | Diagnostic Test: Blood test<br>* Patients in each group will have a blood sample (9x7 mL) for eosinophils isolation, study of activation markers on whole blood and serum biomarkers testing<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Comparison of ECP concentrations in supernatants of eosinophils culture | Eosinophils will be sorted, stimulated in vitro for 2 hours and ECP concentration will be assessed in supernatants | Baseline: one session | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Comparison of median fluorescence intensities of several surface markers on blood eosinophils, or comparison of percentages of positive cells among all eosinophils for a given marker (flow cytometry) | MFI/% of CD69, HLA class II, CD9, CD11c, CD44, CCR3, CRTH2, IL-5R on blood eosinophils | Baseline: one session | 
| Gene expression profiles will be compared between SSc patients and healthy controls (whole transcriptome assay) | Whole genome, transcriptomic approach (differentially expressed genes will be identified using a fold change cutoff) | Baseline: one session | 
| In skin biopsies: density of eosinophils, extracellular ECP and MBP deposits (absent in healthy skin), and density of eotaxin-1-producing cells will be assessed in damaged skin and apparently healthy skin of SSc patients |  | Baseline: one session | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Eosinophil, Fibrosis, Systemic Scleroderma