The Immunomodulatory Signature of Extracellular Vesicles From Cardiosphere-Derived Cells: A Proteomic and miRNA Profiling

Esther López; Federica Marinaro; María de Los Ángeles de Pedro; Francisco Miguel Sánchez-Margallo; María Gómez-Serrano; Viviane Ponath; Elke Pogge von Strandmann; Inmaculada Jorge; Jesús Vázquez; Luis Miguel Fernández-Pereira; Verónica Crisóstomo; Verónica Álvarez; Javier G Casado

doi:10.3389/fcell.2020.00321

The Immunomodulatory Signature of Extracellular Vesicles From Cardiosphere-Derived Cells: A Proteomic and miRNA Profiling

Front Cell Dev Biol. 2020 Jun 9:8:321. doi: 10.3389/fcell.2020.00321. eCollection 2020.

Authors

Esther López¹, Federica Marinaro¹, María de Los Ángeles de Pedro¹, Francisco Miguel Sánchez-Margallo^{1

2}, María Gómez-Serrano^{2

3

4}, Viviane Ponath^{5

6}, Elke Pogge von Strandmann^{5

6}, Inmaculada Jorge^{2

3}, Jesús Vázquez^{2

3}, Luis Miguel Fernández-Pereira⁷, Verónica Crisóstomo^{1

2}, Verónica Álvarez¹, Javier G Casado^{1

2}

Affiliations

¹ Stem Cell Therapy Unit, Jesús Usón Minimally Invasive Surgery Centre, Cáceres, Spain.
² CIBER de Enfermedades Cardiovasculares (CIBERCV), Madrid, Spain.
³ Laboratory of Cardiovascular Proteomics, Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid, Spain.
⁴ Institute of Molecular Biology and Tumor Research (IMT), Center for Tumor Biology and Immunology (ZTI), Philipps University, Marburg, Germany.
⁵ Institute for Tumor Immunology, Center for Tumor Biology and Immunology (ZTI), Philipps University, Marburg, Germany.
⁶ Clinic for Hematology, Oncology, and Immunology, Philipps University, Marburg, Germany.
⁷ Immunology Department, Hospital San Pedro de Alcántara, Cáceres, Spain.

Abstract

Experimental data demonstrated that the regenerative potential and immunomodulatory capacity of cardiosphere-derived cells (CDCs) is mediated by paracrine mechanisms. In this process, extracellular vesicles derived from CDCs (EV-CDCs) are key mediators of their therapeutic effect. Considering the future applicability of these vesicles in human diseases, an accurate preclinical-to-clinical translation is needed, as well as an exhaustive molecular characterization of animal-derived therapeutic products. Based on that, the main goal of this study was to perform a comprehensive characterization of proteins and miRNAs in extracellular vesicles from porcine CDCs as a clinically relevant animal model. The analysis was performed by identification and quantification of proteins and miRNA expression profiles. Our results revealed the presence of clusters of immune-related and cardiac-related molecular biomarkers in EV-CDCs. Additionally, considering that priming stem cells with inflammatory stimuli may increase the therapeutic potential of released vesicles, here we studied the dynamic changes that occur in the extracellular vesicles from IFNγ-primed CDCs. These analyses detected statistically significant changes in several miRNAs and proteins. Notably, the increase in interleukin 6 (IL6) protein, as well as the increase in mir-125b (that targets IL6 receptor) was especially relevant. These results suggest a potential involvement of EV-CDCs in the regulation of the IL6/IL6R axis, with implications in inflammatory-mediated diseases.

Keywords: cardiac stem cells; cardiosphere-derived cells; extracellular vesicles; interferon-γ; miRNA–microRNA; priming; proteomic analyses; quantitative polymerase chain reaction.