Myofibroblast-Derived Exosome Induce Cardiac Endothelial Cell Dysfunction.

Prabhat Ranjan,Rajesh Kumari,Raj Kishore,Jing Li,Suresh Kumar Verma,Harish Pal,Sumanta Kumar Goswami,Zainab Suleiman,Zhongjian Cheng,Prasanna Krishnamurthy

doi:10.3389/fcvm.2021.676267

Prabhat Ranjan, Rajesh Kumari + Show 8 more

Open Access

https://doi.org/10.3389/fcvm.2021.676267

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Abstract

Background: Endothelial cells (ECs) play a critical role in the maintenance of vascular homeostasis and in heart function. It was shown that activated fibroblast-derived exosomes impair cardiomyocyte function in hypertrophic heart, but their effect on ECs is not yet clear. Thus, we hypothesized that activated cardiac fibroblast-derived exosomes (FB-Exo) mediate EC dysfunction, and therefore modulation of FB-exosomal contents may improve endothelial function.Methods and Results: Exosomes were isolated from cardiac fibroblast (FB)-conditioned media and characterized by nanoparticle tracking analysis and electron microscopy. ECs were isolated from mouse heart. ECs were treated with exosomes isolated from FB-conditioned media, following FB culture with TGF-β1 (TGF-β1-FB-Exo) or PBS (control) treatment. TGF-β1 significantly activated fibroblasts as shown by increase in collagen type1 α1 (COL1α1), periostin (POSTN), and fibronectin (FN1) gene expression and increase in Smad2/3 and p38 phosphorylation. Impaired endothelial cell function (as characterized by a decrease in tube formation and cell migration along with reduced VEGF-A, Hif1α, CD31, and angiopoietin1 gene expression) was observed in TGF-β1-FB-Exo treated cells. Furthermore, TGF-β1-FB-Exo treated ECs showed reduced cell proliferation and increased apoptosis as compared to control cells. TGF-β1-FB-Exo cargo analysis revealed an alteration in fibrosis-associated miRNAs, including a significant increase in miR-200a-3p level. Interestingly, miR-200a-3p inhibition in activated FBs, alleviated TGF-β1-FB-Exo-mediated endothelial dysfunction.Conclusions: Taken together, this study demonstrates an important role of miR-200a-3p enriched within activated fibroblast-derived exosomes on endothelial cell biology and function.

Highlights

The endothelium maintains normal vascular tone and blood fluidity under normal homeostatic conditions
Taken together, this study demonstrates an important role of miR-200a-3p enriched within activated fibroblast-derived exosomes on endothelial cell biology and function
Real time qRT-PCR data revealed that collagen type 1, α1 (Figure 1A), periostin (Figure 1B) and fibronectin (Figure 1C) mRNA expression was significantly increased in transforming growth factor-β (TGFβ)-treated Neonatal rat ventricular fibroblasts (NRVF) compared to bovine serum albumin (BSA) treatment

Summary

Introduction

The endothelium maintains normal vascular tone and blood fluidity under normal homeostatic conditions. Transforming growth factor beta-1 (TGF-β1), a profibrotic cytokine, plays an important role in profibrotic signaling, and fibrosis in the heart [9, 10]. It enhances production of extracellular matrix proteins by fibroblasts. As fibrosis progresses, it leads to tissue damage and inflammation [11]. In this process, endothelial cells lose their cellular integrity and undergo endothelial to mesenchymal transition (EndMT) [12, 13]. Endothelial cells (ECs) play a critical role in the maintenance of vascular homeostasis and in heart function. We hypothesized that activated cardiac fibroblast-derived exosomes (FB-Exo) mediate EC dysfunction, and modulation of FB-exosomal contents may improve endothelial function

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Conclusion