Atherosclerotic Pre-Conditioning Affects the Paracrine Role of Circulating Angiogenic Cells Ex-Vivo

Int J Mol Sci . 2020 Jul 24;21(15):5256. doi: 10.3390/ijms21155256.

Fecha de la publicación: 24/07/2020

Autor: Sara Eslava-Alcon (1, 2), Mª Jesús Extremera-García (1, 2), Ismael Sanchez-Gomar (1, 2), Lucía Beltrán-Camacho (1, 2), Antonio Rosal-Vela (1, 2), Javier Muñoz (3, 4), Nuria Ibarz (3, 4), Jose Angel Alonso-Piñero (1, 2), Marta Rojas-Torres (1, 2), Margarita Jiménez-Palomares (1, 2), Almudena González-Rovira (1, 2), Rosario Conejero (5), Esther Doiz (5), Manuel Rodriguez-Piñero (5), Rafael Moreno-Luna (6), Mª Carmen Durán-Ruiz (1, 2)

Palabras clave: angiogenesis, atherosclerotic factors, circulating angiogenic cells, endothelial colony forming cells, paracrine role, proteomics; secretome



1Biomedicine, Biotechnology and Public Health Department, Cádiz University, 11002 Cádiz, Spain.

2Institute of Research and Innovation in Biomedical Sciences Cadiz (INIBICA), 11009 Cádiz, Spain.

3Proteomics Unit, Spanish National Cancer Research Centre (CNIO), 28029 Madrid, Spain.

4ISCIII-ProteoRed, 28029, 28029 Madrid, Spain.

5Angiology& Vascular Surgery Unit, Hospital Universitario Puerta del Mar, 11009 Cádiz, Spain.

6Laboratory of Neuroinflammation, Hospital Nacional de Paraplejicos, SESCAM, 45071 Toledo, Spain.


In atherosclerosis, circulating angiogenic cells (CAC), also known as early endothelial progenitor cells (eEPC), are thought to participate mainly in a paracrine fashion by promoting the recruitment of other cell populations such as late EPC, or endothelial colony-forming cells (ECFC), to the injured areas. There, ECFC replace the damaged endothelium, promoting neovascularization. However, despite their regenerative role, the number and function of EPC are severely affected under pathological conditions, being essential to further understand how these cells react to such environments in order to implement their use in regenerative cell therapies. Herein, we evaluated the effect of direct incubation ex vivo of healthy CAC with the secretome of atherosclerotic arteries. By using a quantitative proteomics approach, 194 altered proteins were identified in the secretome of pre-conditioned CAC, many of them related to inhibition of angiogenesis (e.g., endostatin, thrombospondin-1, fibulins) and cell migration. Functional assays corroborated that healthy CAC released factors enhanced ECFC angiogenesis, but, after atherosclerotic pre-conditioning, the secretome of pre-stimulated CAC negatively affected ECFC migration, as well as their ability to form tubules on a basement membrane matrix assay. Overall, we have shown here, for the first time, the effect of atherosclerotic factors over the paracrine role of CAC ex vivo. The increased release of angiogenic inhibitors by CAC in response to atherosclerotic factors induced an angiogenic switch, by blocking ECFC ability to form tubules in response to pre-conditioned CAC. Thus, we confirmed here that the angiogenic role of CAC is highly affected by the atherosclerotic environment.