Doxorubicin upregulates CXCR4 via miR-200c/ZEB1-dependent mechanism in human cardiac mesenchymal progenitor cells

Sara Beji,Yuri D'Alessandra,Matteo Bertolotti,Alessandra Magenta,Maurizio C Capogrossi,Chiara Cencioni,Sarah Pizzolato,Lucia Cicchillitti,Elisa Gambini,Daniele Avitabile,Giulio Pompilio,Gabriella Spaltro,Giuseppina Milano,Alessandro Scopece,Angela Raucci,Mario Picozza,Giulia Piaggio

doi:10.1038/cddis.2017.409

Abstract

Doxorubicin (DOXO) treatment is limited by its cardiotoxicity, since it causes cardiac-progenitor-cell depletion. Although the cardioprotective role of the stromal cell-derived factor-1/C-X-C chemokine receptor type 4 (SDF1/CXCR4) axis is well established, its involvement during DOXO-induced cardiotoxicity has never been investigated. We showed that in a mouse model of DOXO-induced cardiomyopathy, CXCR4+ cells were increased in response to DOXO, mainly in human cardiac mesenchymal progenitor cells (CmPC), a subpopulation with regenerative potential. Our in vitro results showed a CXCR4 induction after 24 h of DOXO exposure in CmPC. SDF1 administration protected from DOXO-induced cell death and promoted CmPC migration. CXCR4 promoter analysis revealed zinc finger E-box binding homeobox 1 (ZEB1) binding sites. Upon DOXO treatment, ZEB1 binding decreased and RNA-polymerase-II increased, suggesting a DOXO-mediated transcriptional increase in CXCR4. Indeed, DOXO induced the upregulation of miR-200c, that directly targets ZEB1. SDF1 administration in DOXO-treated mice partially reverted the adverse remodeling, decreasing left ventricular (LV) end diastolic volume, LV ejection fraction and LV anterior wall thickness in diastole, recovering LV end systolic pressure and reducing±dP/dt. Moreover, in vivo administration of SDF1 partially reverted DOXO-induced miR-200c and p53 protein upregulation in mouse hearts. In addition, downmodulation of ZEB1 mRNA and protein by DOXO was significantly increased by SDF1. In keeping, p21 mRNA, that is induced by p53 and inhibited by ZEB1, is induced by DOXO treatment and is decreased by SDF1 administration. This study showed new players of the DOXO-induced cardiotoxicity, that can be exploited to ameliorate DOXO-associated cardiomyopathy.

Highlights

Among them, Doxorubicin (DOXO) is largely used in different types of tumors, including breast cancer, esophageal carcinoma, osteosarcoma, sarcomas and lymphomas.[1]
The cardioprotective role of the SDF1/CXCR4 axis was already established in myocardial infarction,[35,36,37,38] it was never studied in DOXO-induced cardiotoxicity
We observed that DOXO induces the upregulation of CXCR4, making human cardiac mesenchymal progenitor cells (CmPC) more prone to respond to SDF1 stimulation

Summary

Introduction

Doxorubicin (DOXO) is largely used in different types of tumors, including breast cancer, esophageal carcinoma, osteosarcoma, sarcomas and lymphomas.[1]. SDF1 provides trophic support for cells, stimulates progenitor cell differentiation and promotes angiogenesis through a paracrine mechanism.[14] the activation of the SDF1/CXCR4 axis promotes extensive mobilization of CmPC and supports cardiac repair of the infarcted heart.[15,16,17] Notably, the cardiac protective role of this axis has been recently confirmed in a clinical setting of ischemic heart failure.[18] in dilated cardiomyopathy, SDF1 increases and enhances the number of circulating progenitor cells[19] and DOXO-induced cardiomyopathy promotes mesenchymal stem cell migration to the heart, where SDF1 expression is elevated.[20]. MicroRNAs (miRNAs) are 21–23 nucleotides RNA molecules that regulate the stability or translational efficiency of target messenger RNAs.[21] miRNAs control a wide range of cell functions and have been associated with inflammation, oxidative stress and different pathologies, including heart failure, cardiac hypertrophy and myocardial arrhythmias.[22,23]

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Conclusion