An iPSC-derived bio-inspired scaffold modelling the structure and the effects of extracellular matrix in cardiac fibrosis

ABSTRACT: Cardiac fibrosis occurs following insults to the myocardium and is characterized by the abnormal accumulation of non-compliant extracellular matrix (ECM), which compromises cardiomyocyte (CMs) contractile activity and eventually leads to heart failure. This phenomenon is driven by the differentiation of cardiac fibroblasts (cFbs) into myofibroblasts and results in changes in ECM biochemical and structural properties. The lack of predictive in vitro models of heart fibrosis has so far hampered the search for innovative treatments. Here, we adopted a protocol to generate cFbs from human induced pluripotent stem cells (iPSCs) and activate them to a myofibroblast phenotype by tuning basic fibroblast growth factor (bFGF) and transforming growth factor beta 1 (TGF-β) signalling. We next confirmed that TGF-β stimulation prompted iPSC-derived cells to acquire key features of myofibroblasts, like SMAD2/3 nuclear shuttling, the formation of aligned alpha-smooth muscle actin (α-SMA)-rich stress fibres and increased focal adhesions (FAs) assembly. Additionally, we devised a single-step decellularization protocol to obtain and thoroughly characterize the biochemical and mechanical properties of the ECM secreted by activated cFbs. After revealing that iPSC-derived myofibroblasts secrete an abundant, collagen-rich and stiff ECM characterized by distinct viscoelastic properties, we demonstrated that this pro-fibrotic ECM activates mechanosensitive pathways in iPSC-derived CMs (iPSC-CMs), impacting their shape, sarcomere length, phenotype and calcium handling properties. We thus propose these human bio-inspired matrices as animal-free, isogenic CM culture substrates recapitulating key pathophysiological changes occurring at the cellular level during cardiac fibrosis.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Extracellular Matrix

SUBMITTER: Václav Pustka  

LAB HEAD: Zbynek Zdrahal

PROVIDER: PXD049208 | Pride | 2025-05-06

REPOSITORIES: Pride