The Microenvironment of Decellularized Extracellular Matrix from Heart Failure Myocardium Alters the Balance between Angiogenic and Fibrotic Signals from Stromal Primitive Cells.

Ciro Maiello,Elisa Messina,Stefania Montagnani,Veronica Romano,Franca Di Meglio,Immacolata Belviso,Giacomo Frati,Vittorio Picchio,Daria Nurzynska,Francesca Pagano,Francesco Angelini,Cristina Nocella,Anna Maria Sacco,Isotta Chimenti,Clotilde Castaldo

doi:10.3390/ijms21217903

Abstract

Cardiac adverse remodeling is characterized by biological changes that affect the composition and architecture of the extracellular matrix (ECM). The consequently disrupted signaling can interfere with the balance between cardiogenic and pro-fibrotic phenotype of resident cardiac stromal primitive cells (CPCs). The latter are important players in cardiac homeostasis and can be exploited as therapeutic cells in regenerative medicine. Our aim was to compare the effects of human decellularized native ECM from normal (dECM-NH) or failing hearts (dECM-PH) on human CPCs. CPCs were cultured on dECM sections and characterized for gene expression, immunofluorescence, and paracrine profiles. When cultured on dECM-NH, CPCs significantly upregulated cardiac commitment markers (CX43, NKX2.5), cardioprotective cytokines (bFGF, HGF), and the angiogenesis mediator, NO. When seeded on dECM-PH, instead, CPCs upregulated pro-remodeling cytokines (IGF-2, PDGF-AA, TGF-β) and the oxidative stress molecule H2O2. Interestingly, culture on dECM-PH was associated with impaired paracrine support to angiogenesis, and increased expression of the vascular endothelial growth factor (VEGF)-sequestering decoy isoform of the KDR/VEGFR2 receptor. Our results suggest that resident CPCs exposed to the pathological microenvironment of remodeling ECM partially lose their paracrine angiogenic properties and release more pro-fibrotic cytokines. These observations shed novel insights on the crosstalk between ECM and stromal CPCs, suggesting also a cautious use of non-healthy decellularized myocardium for cardiac tissue engineering approaches.

Highlights

Despite remarkable progress in terms of early diagnosis and prevention, heart failure (HF) is still a leading cause of death in Western countries [1], with ischemic heart disease (IHD) being a main etiological cause with its multifaceted pathogenetic mechanisms [2]
Decellularized native extracellular matrix sections obtained from normal or pathological hearts with advanced HF were seeded with human cardiac stromal primitive cells (CPCs) isolated from IHD patients (Figure 1A,B) and cultured up to 7 days, with standard culture on fibronectin (FN)-coating as control
Revealed that the proportion of transcript for membrane-associated KDR/VEGFR2 was negligible in all conditions, while its soluble form was the absolute majority in all conditions (Figure 5)

Summary

Introduction

Despite remarkable progress in terms of early diagnosis and prevention, heart failure (HF) is still a leading cause of death in Western countries [1], with ischemic heart disease (IHD) being a main etiological cause with its multifaceted pathogenetic mechanisms [2]. IHD affects the viability and function of cardiomyocytes [3] and other resident cells (e.g., smooth muscle, endothelial, and stromal), leading to significant changes in the composition and architecture of the extracellular matrix (ECM). The crosstalk between ECM and resident cells in IHD and HF produces a detrimental microenvironment, which impacts on all myocardial cells [4]. Cardiac regenerative medicine, including cardiac cell therapy (CCT) [5,6,7], aims at achieving regeneration for compensating parenchymal loss and recovering heart function. In the context of CCT, regenerative cells need a supporting microenvironment (for example in terms of ECM) to optimize this crosstalk, to boost regeneration and angiogenesis, while counteracting fibrosis [9,10,11]

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