Tissue Modeling in Systemic Sclerosis Using Induced Pluripotent Stem Cells (iPSCs)

Condition(s) studied

Investigational drug(s) / intervention(s)

Blood Sample Collection: Collection of approximately 28 mL of blood from each participant to generate induced pluripotent stem cell (iPSC) lines for in vitro cellular and tissue modeling analyses.

Study 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.

Eligibility

Primary outcome measure(s)

• Rate of successful iPSC line generation from PBMCs in diffuse/severe systemic sclerosis patients — 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 — 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 — 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 — 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

Trial sites (1)

More University Hospital, Montpellier trials in France

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