Kidney Week

Abstract: SA-PO981

Senescent MVs from IS-Treated Endothelial Cells Induce Vascular Calcification

Session Information

• Hypertension and CVD: Mechanisms - II
  October 27, 2018 | Location: Exhibit Hall, San Diego Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Hypertension and CVD

• 1403 Hypertension and CVD: Mechanisms

Authors

• Alique, Matilde, Universidad de Alcalá, Alcalá de Henares, Spain

Matilde Alique,

• Carracedo, Julia, Complutense University, MADRID., Spain

Julia Carracedo,

• Bodega, Guillermo, Universidad de Alcalá, Alcalá de Henares, Spain

Guillermo Bodega,

• Corchete, Elena, Hospital Universitario Fundación Alcorcon, Alcorcon (Madrid), Spain

Elena Corchete,

• Garcia menendez, Estefanya, Hospital Puerta de Hierro Majadahonda, Majadahonda, Spain

Estefanya Garcia menendez,

• De Sequera, Patricia, University Hospital Infanta Leonor, MADRID, Spain

Patricia De Sequera,

• Marques Vidas, Maria, Hospital Universitario Puerta de Hierro Majadahonda, Majadahonda, MAD, Spain

Maria Marques Vidas,

• Perez-Garcia, Rafael, Hospital Infanta Leonor, Madrid, Spain

Rafael Perez-Garcia,

• Portolés, J.M., Hospital Puerta De Hierro, Majadahonda (Madrid), MAD, Spain

J.M. Portolés,

• Ramirez, Rafael, Universidad de Alcalá, Alcalá de Henares, Spain

Rafael Ramirez,

Background

Vascular calcification (VC) is a premature event of cardiovascular inflammatory diseases (CVD) in patients with chronic kidney disease (CKD). But identifying patients at risk of developing VC is complex, and it is necessary to expand the knowledge of the initial events that allow the progression of this pathology. Recently, we reported that senescent endothelial cells produce microvesicles (MV) which can act triggering the development of VC. As induction of premature endothelial senescence is a physiopathological mechanism in CVD associated with CKD, the objective of this study is to investigate whether MVs from endothelial cells that undergo premature senescence induced by the uremic toxin indoxyl sulfate (IS) can also modify vascular smooth muscle cells by making them susceptible to bone transformation. Besides, we study the mechanism(s) involved in VC promotion.

Methods

Endothelial cells (HUVEC) were treated with IS (250 µM; 4 days) and senescence was quantified with the β-galactosidase kit. The MV produced by these senescent HUVECs were isolated and subsequently characterized by flow cytometry and used for the treatment of vascular smooth muscle cells (HASMC). In HASMC, the calcium deposits were evaluated by alizarin red. At 30 days, the calcium content was quantified by the phenolsulfonephthalein kit. Expression of pro-calcifying genes was determined in HASMC by quantitative PCR. Western blot established the expression of a specific HASMC marker.

Results

IS-treated induced early senescence in HUVEC (control: 7.07±2.87% vs. IS: 72.90±13.09%, p <0.0001) and an increase in the production of MV was observed (control: 1.68±1.00 vs. IS: 9.92±4.29 MV/cell, p <0.005). Senescent MVs from IS-treated endothelial cells induced calcification in HASMC associated with deregulation in the expression of procalcifying genes (Runx2, BMP2). Also, the MV generated in HUVEC treated with IS produce a dedifferentiation process of the HASMC (SM22α expression).

Conclusion

MV produced by endothelial cells which develop early senescence as a consequence of the stress generated by the uremic toxins promote the development of VC. These results may be useful to develop biomarkers and therapeutic tools to prevent or treated patients with CVD and/or CKD at risk of developing VC.