Normal versus Pathological Cardiac Fibroblast-Derived Extracellular Matrix Differentially Modulates Cardiosphere-Derived Cell Paracrine Properties and Commitment

Abstract

Human resident cardiac progenitor cells (CPCs) isolated as cardiosphere-derived cells (CDCs) are under clinical evaluation as a therapeutic product for cardiac regenerative medicine. Unfortunately, limited engraftment and differentiation potential of transplanted cells significantly hamper therapeutic success. Moreover, maladaptive remodelling of the extracellular matrix (ECM) during heart failure progression provides impaired biological and mechanical signals to cardiac cells, including CPCs. In this study, we aimed at investigating the differential effect on the phenotype of human CDCs of cardiac fibroblast-derived ECM substrates from healthy or diseased hearts, named, respectively, normal or pathological cardiogel (CG-N/P). After 7 days of culture, results show increased levels of cardiogenic gene expression (NKX2.5, CX43) on both decellularized cardiogels compared to control, while the proportion and staining patterns of GATA4, OCT4, NKX2.5, ACTA1, VIM, and CD90-positive CPCs were not affected, as assessed by immunofluorescence microscopy and flow cytometry analyses. Nonetheless, CDCs cultured on CG-N secreted significantly higher levels of osteopontin, FGF6, FGF7, NT-3, IGFBP4, and TIMP-2 compared to those cultured on CG-P, suggesting overall a reduced trophic and antiremodelling paracrine profile of CDCs when in contact with ECM from pathological cardiac fibroblasts. These results provide novel insights into the bidirectional interplay between cardiac ECM and CPCs, potentially affecting CPC biology and regenerative potential.

Decellularized Extracellular Matrix Materials for Cardiac Repair and Regeneration

“…In addition to tissue derived dECM, there have been studies on cardiac‐specific dECM derived from cardiac fibroblasts grown in sheets, called fibroblast‐derived matrix (FDM) . These scaffolds can be used as patches, either directly or with additional cell seeing.…”

“…In addition, by altering crosslinking of the FDM to increase stiffness, there were further increases in cardiogenic differentiation, with α5 integrin expression increased on crosslinked compared to non‐crosslinked FDM. In a cardiac‐specific method, Pagano et al grew CPCs isolated as cardiosphere‐derived cells (CDCs) on FDM derived from fibroblasts harvested from healthy and infarcted hearts . After 7 d in culture, both systems showed expression of cardiac differentiation, although CDCs on healthy FDM secreted higher levels of reparative factors such as osteopontin, FGF, IGF, and TIMP‐2, modeling the decrease in reparative and antiremodeling potential of CDCs when injected in an infarct environment.…”

Cardiac Progenitor Cells: The Matrix Has You

“…Despite the synergistic effect of all ECM components, a direct supporting mechanism has been already reported for some ECM molecules like fibronectin, proven to be necessary for endogenous CPC proliferation and activation for repair through focal adhesion signaling . The ECM's influence on CPCs, though, may act at multiple levels: we have previously reported that human CPCs cultured short‐term on cardiac fibroblast‐derived ECM substrates from failing hearts release less trophic and anti‐remodeling paracrine factors, such as tissue inhibitor of metalloproteinases 2 (TIMP2), compared to substrates from fibroblasts from healthy hearts . Moreover, multiple studies have investigated several combinations of ECM protein substrates (e.g., fibronectin, laminin, collagen) with variable mechanical stimuli (e.g., cyclic strain), suggesting that mechanical and biological signals can compensate and modulate each other in different proportions , making it very difficult to design an artificial matrix from scratch.…”

Epicardial Transplantation of Cardiac Progenitor Cells Based Cells Sheets is More Promising Method for Stimulation of Myocardial Regeneration, Than Conventional Cell Injections

“…Кроме того, именно состояние белков матрикса определяет регенеративные свойства ПКС. Культивирование ПКС на матриксе, наработанном клетками пациентов с хронической сердечной недостаточностью, вызывает подавление их паракринной активности по сравнению с матриксом от здоровых доноров [27]. Более того, в нескольких исследованиях продемонстрировано, что соотношение компонентов внеклеточного матрикса и их модификация являются определяющим фактором для запуска регенеративной программы в прогениторных клетках [28,29].…”