Extracellular Ubiquitin Increases Expression of Angiogenic Molecules and Stimulates Angiogenesis in Cardiac Microvascular Endothelial Cells

Authors

Rebecca J. Steagall, Christopher R. Daniels, Suman Dalal, William L. Joyner, Mahipal Singh, Krishna Singh

Abstract

Objective
Extracellular ubiquitin (Ub) is an immune modulator that plays a role in suppression of inflammation, organ injury, myocyte apoptosis and fibrosis. The purpose of this study was to investigate the effects of extracellular Ub on the process of cardiac angiogenesis.
Methods
Cardiac microvascular endothelial cells (CMECs) and aortic tissue were isolated from rats to measure changes in angiogenic protein levels and to assess angiogenic responses to extracellular Ub.
Results
In CMECs, extracellular Ub increased protein levels of vascular endothelial growth factor-A (VEGF-A) and matrix metalloproteinase-2 (MMP-2), known angiogenesis regulators. CMECs demonstrated enhanced rearrangement of fibrillar actin and migration in response to Ub treatment. Ub-treated CMECs demonstrated an increase in tube network formation which was inhibited by the CXCR4 receptor antagonist, AMD3100. Methylated Ub, unable to form polyubiquitin chains, enhanced tube network formation. Aortic ring sprouting assays demonstrated that Ub increases microvessel sprouting in the Matrigel.
Conclusion
The results of our study suggest a novel role for extracellular Ub in cardiac angiogenesis, providing evidence that extracellular Ub, at least in part acting via the CXCR4 receptor, has the potential to facilitate the process of angiogenesis in myocardial endothelial cells.