Hyaluronan Mixed Esters of Butyric and Retinoic Acid Drive Cardiac and Endothelial Fate in Term Placenta Human Mesenchymal Stem Cells and Enhance Cardiac Repair in Infarcted Rat Hearts

Carlo Ventura,Silvia Cantoni,Francesca Bianchi,Vincenzo Lionetti,Claudia Cavallini,Ignazio Scarlata,Laura Foroni,Margherita Maioli,Laura Bonsi,Francesco Alviano,Valentina Fossati,Gian Paolo Bagnara,Gianandrea Pasquinelli,Fabio A Recchia,Alberto Perbellini

doi:10.1074/jbc.m609350200

Abstract

We have developed a mixed ester of hyaluronan with butyric and retinoic acid (HBR) that acted as a novel cardiogenic/vasculogenic agent in human mesenchymal stem cells isolated from bone marrow, dental pulp, and fetal membranes of term placenta (FMhMSCs). HBR remarkably enhanced vascular endothelial growth factor (VEGF), KDR, and hepatocyte growth factor (HGF) gene expression and the secretion of the angiogenic, mitogenic, and antiapoptotic factors VEGF and HGF, priming stem cell differentiation into endothelial cells. HBR also increased the transcription of the cardiac lineage-promoting genes GATA-4 and Nkx-2.5 and the yield of cardiac markerexpressing cells. These responses were notably more pronounced in FMhMSCs. FMhMSC transplantation into infarcted rat hearts was associated with increased capillary density, normalization of left ventricular function, and significant decrease in scar tissue. Transplantation of HBR-preconditioned FMhM-SCs further enhanced capillary density and the yield of human vWF-expressing cells, additionally decreasing the infarct size. Some engrafted, HBR-pretreated FMhMSCs were also positive for connexin 43 and cardiac troponin I. Thus, the beneficial effects of HBR-exposed FMhMSCs may be mediated by a large supply of angiogenic and antiapoptotic factors, and FMhMSC differentiation into vascular cells. These findings may contribute to further development in cell therapy of heart failure.

Highlights

Loss of cardiomyocytes due to myocardial infarction or hereditary cardiomyopathies may represent causative factors in the progression toward heart failure
Flow cytometry revealed that BMhMSCs, DPhMSCs, and FMhMSCs were positively stained with SH2, which identifies an epitope of endoglin (CD105), the transforming growth factor-␤ receptor III present on endothelial cells, erythroblasts, monocytes, and connective tissue stromal cells, facilitating the enrichment of stromal progenitors from bone marrow [29]
Like BMhMSCs, DPhMSCs and FMhMSCs were recognized by the SH3 and SH4 antibodies, which identify epitopes on culture-expanded stromal cells and bind CD73, a molecule involved in B-cell activation [30]

Summary

Introduction

Loss of cardiomyocytes due to myocardial infarction or hereditary cardiomyopathies may represent causative factors in the progression toward heart failure. The initial observations and the scientific underpinning of the human trials have been challenged by a number of studies using mouse bone marrow hematopoietic cells expressing LacZ or green fluorescent protein to enable detection in recipient hearts In these studies little or no reporter activity could be seen after direct stem cell injection into the damaged heart or continuous cell transfusion into the bloodstream of infarcted mice [3, 4]. These controversies prompt further investigations assessing whether cell populations retaining a therapeutic potential for damaged hearts may be identified in human bone marrow or alternative sources. We investigated whether HBR may commit BMhMSCs, DPhMSCs, or FMhMSCs into cardiovascular lineages and/or enhance their potential as trophic mediators and whether, in the affirmative, transplantation of HBR-pretreated hMSCs into infarcted rat hearts may result in enhanced cardiovascular repair

Methods

Results

Conclusion