Tissue Modeling in Systemic Sclerosis Using Induced Pluripotent Stem Cells (iPSCs)
Trial Snapshot
- Phase
- Not Applicable
- Status
- Not yet recruiting
- Enrollment
- 16
- Locations
- 1
- Primary Endpoint
- Rate of successful iPSC line generation from PBMCs in diffuse/severe systemic sclerosis patients
Study Overview
Brief Summary
The objective of the study is to establish an in vitro model using iPSCs to test the hypotheses developed.
The primary objective is to generate, via the SAFE-IPS platform, 8 iPSC lines derived from blood samples taken from:
- Two patients with severe/diffuse SSc with multi-organ involvement (with anti-SCl-70 autoantibodies)
- Two of their healthy close relatives
- Two patients with uncomplicated SSc with localized involvement (with non-SCl-70 autoantibodies)
- Two of their healthy close relatives
These 8 cell lines will be differentiated by several teams at the FHU REGENHAB into:
- Immune cells (monocytes/macrophages, neutrophils, dendritic cells, B/T lymphocytes)
- Mesenchymal stromal cells
- Myocardial cells
- Skin cells (fibroblasts)
- Synovial cells
- Bronchial cells
- Endothelial cells
This project aims to map cellular and tissue heterogeneity using iPSC lines obtained from patients with both severe and mild SSc, employing single-cell RNA-seq under various pathological conditions, with and without autologous (or even heterologous) autoimmune stimulation. For example, the percentages of different cell types comprising a tissue in these various situations will be calculated. Comparisons will be made with control groups using healthy iPSC lines by recruiting healthy subjects with the same genetic profile and gender as the patients.
Detailed Description
Background:
Systemic sclerosis (SSc) is a rare autoimmune connective tissue disease that predominantly affects women, often has a severe course, and is characterized by heterogeneous multiorgan involvement. Its pathophysiology, involving microvascular abnormalities, autoimmune activation, and progressive fibrosis, remains incompletely understood, particularly regarding its variations across clinical forms and affected organs. Currently available treatments rely primarily on immunosuppressive strategies, which have limited efficacy and are associated with significant adverse effects, with no therapeutic option available to prevent certain major complications. In this context of unmet medical need, the use of induced pluripotent stem cells (iPSCs) derived from patients with SSc represents an innovative approach to modeling the pathophysiological mechanisms of the disease in vitro. The FHU Regenhab consortium's demonstrated ability to differentiate iPSCs into several relevant cell types offers a unique opportunity to study specific organ damage and identify new therapeutic targets, paving the way for more personalized treatment strategies.
Objectives:
Primary objective:
The primary objective is to generate, using the SAFE-IPS platform, 8 iPSC lines derived from cells obtained from a blood sample from:
Study Design
- Study Type
- Interventional
- Allocation
- Na
- Intervention Model
- Single Group
- Primary Purpose
- Basic Science
- Masking
- None
Eligibility Criteria
- Ages
- 18 Years to 85 Years (Adult, Older Adult)
- Sex
- All
- Accepts Healthy Volunteers
- Yes
Inclusion Criteria
- •Age between 18 and 85 years
- •Diagnostic criteria for the three groups:
- •> Severe SSc group:
- •Diagnosis of diffuse/severe systemic sclerosis by a physician for at least 12 months based on recommendations (according to ACR criteria)
- •Known positivity of autoimmunity directed against Scl-70
- •> Localized/non-complicated SSc group:
- •Diagnosis of systemic sclerosis by a physician for at least 12 months based on recommendations (according to ACR criteria)
- •Documentation of the absence of major vital organ involvement
- •Negative anti-Scl-70 autoantibodies but presence of other systemic sclerosis-related autoantibodies
- •> Healthy subjects group:
- •First-degree relative of a patient recruited in one of the systemic sclerosis groups (father, mother, brother, sister, adult child)
- •Absence of systemic autoimmune diseases
Exclusion Criteria
- •Other diseases that may affect erythroid progenitor cells (non-exhaustive: active cancer, hematological malignancy, chemotherapy targeting DNA)
- •Patient in an exclusion period determined by another protocol
- •Protected populations according to French public health law (pregnant or breastfeeding women; individuals deprived of liberty by judicial or administrative decision; adults under legal protection (any form of guardianship))
- •Absence of informed consent
- •Not affiliated with a national health insurance system
Arms & Interventions
Participants with Systemic Sclerosis and Healthy Controls
Participants undergo blood sample collection for generation of induced pluripotent stem cells (iPSCs) and in vitro cellular and tissue modeling.
Intervention: Blood Sample Collection (Procedure)
Outcomes
Primary Outcomes
Rate of successful iPSC line generation from PBMCs in diffuse/severe systemic sclerosis patients
Time Frame: At inclusion (Day 0 blood draw; iPSC generation assessed within approximately 3 months post-collection)
Percentage of participants with diffuse/severe systemic sclerosis (anti-Scl-70 positive) from whom at least one iPSC line is successfully generated via Sendai virus reprogramming of peripheral blood mononuclear cells (PBMCs). Unit of Measure: % of participants with at least one validated iPSC line
Rate of successful differentiation of iPSCs into at least one target functional cell type in localized/non-complicated systemic sclerosis patients
Time Frame: At inclusion (assessed within approximately 6 months post-collection)
Among validated iPSC lines from localized/non-complicated SSc patients, the percentage of lines achieving successful directed differentiation into at least one target cell type (cardiomyocytes, macrophages, bronchial epithelial cells, immune cells, or fibroblasts). Success is confirmed by cell-type-specific marker expression assessed by immunofluorescence and flow cytometry. Unit of Measure: % of validated iPSC lines successfully differentiated into ≥1 target cell type
Rate of successful iPSC line generation from PBMCs in localized/non-complicated systemic sclerosis patients
Time Frame: At inclusion (Day 0 blood draw; iPSC generation assessed within approximately 3 months post-collection)
Percentage of participants with localized/non-complicated systemic sclerosis (anti-Scl-70 negative, other SSc-specific antibody positive) from whom at least one iPSC line is successfully generated via Sendai virus reprogramming of PBMCs. Unit of Measure: % of participants with at least one validated iPSC line
Rate of successful differentiation of iPSCs into at least one target functional cell type in diffuse/severe systemic sclerosis patients
Time Frame: At inclusion (assessed within approximately 6 months post-collection)
Among validated iPSC lines from diffuse/severe SSc patients, the percentage of lines achieving successful directed differentiation into at least one target cell type (cardiomyocytes, macrophages, bronchial epithelial cells, immune cells, or fibroblasts). Success is confirmed by cell-type-specific marker expression assessed by immunofluorescence and flow cytometry (e.g., cTnT for cardiomyocytes; CD68/CSFR1 for macrophages; NKX2.1/MUC5AC for bronchial epithelium). Unit of Measure: % of validated iPSC lines successfully differentiated into ≥1 target cell type
Secondary Outcomes
- Change in cardiomyocyte differentiation efficiency following autologous autoantibody exposure(At inclusion)
- Change in macrophage differentiation and inflammatory cytokine secretion following autologous autoantibody exposure(At inclusion)
- Functional characterization of iPSC-derived cardiomyocytes - spontaneous beating activity(At inclusion)
- Molecular characterization of iPSC-derived cardiomyocytes - cardiac marker expression(At inclusion)
- Phenotypic characterization of iPSC-derived macrophages - pan-macrophage surface marker expression(At inclusion)
- Transcriptomic characterization of iPSC-derived macrophages - inflammatory gene regulation(At inclusion)
- Characterization of iPSC-derived immune cells - antigen-presenting cell differentiation efficiency(At inclusion)
- Characterization of iPSC-derived bronchial epithelial cells - stepwise differentiation efficiency(At inclusion)
- Functional characterization of iPSC-derived bronchial air-liquid interface epithelium (iALI) - barrier integrity and mucociliary function(At inclusion)