Comparative transcriptomic analysis validates iPSC derived in-vitro progressive fibrosis model as a screening tool...

Area of Science:

• Biomedical Engineering
• Stem Cell Biology
• Rheumatology

Background:

• Systemic sclerosis (SSc) is a challenging autoimmune disease characterized by fibrosis, impacting research due to limited disease models.
• Studying SSc requires models that capture its progressive nature and cellular plasticity.
• Pluripotent stem cell-derived models are crucial for understanding fibrotic heterogeneity and screening antifibrotic drugs.

Purpose of the Study:

• To develop and characterize a novel in vitro model for studying fibrosis in systemic sclerosis.
• To validate the utility of this model for investigating early and late-stage fibrotic processes.
• To assess the model's potential as a platform for antifibrotic drug screening.

Main Methods:

• Generation of a scar-like phenotype using induced pluripotent stem cell-derived mesenchymal cells (iSCAR) cultured on a wound-mimicking hydrogel.
• RNA sequencing (RNA-seq) to profile transcriptomes at early (48h) and late (13d) stages of iSCAR culture.
• Comparative transcriptomic analysis against a systemic sclerosis (SSc) compendium to identify common fibrotic genes and pathways.

Main Results:

• The iSCAR model successfully recapitulated key "scar-associated" genes (92% early, 85% late).
• Early-stage genes were linked to hypoxia, vascular development, and glycolysis; late-stage genes to senescence.
• The model demonstrated efficacy in validating two antifibrotic compounds, one targeting late-stage fibrosis and another effective in both early and late stages.

Conclusions:

• The iSCAR model provides a robust in vitro platform for studying the complex mechanisms of fibrosis in systemic sclerosis.
• This model effectively captures both early and late-stage fibrotic phenotypes, including associated gene expression.
• The iSCAR system shows significant promise as a tool for screening and validating novel antifibrotic therapies.