@misc{10481/108639,
year = {2025},
month = {12},
url = {https://hdl.handle.net/10481/108639},
abstract = {Platelets and their extracellular vesicles (EVs) have emerged as promising liquid biopsy
biosources for cancer detection and monitoring. The megakaryoblastic MEG-01 cell line
offers a controlled system for generating platelet-like particles (PLPs) and EVs through
valproic-acid-induced differentiation. Here, we performed comprehensive characterization
and proteomic validation of MEG-01-derived populations, native human platelets, and
their EVs using nanoparticle tracking analysis, transmission electron microscopy, imaging
flow cytometry and quantitative proteomics. MEG-01 megakaryocytic differentiation is
characterized by polylobulated nuclei, proplatelet formation, and elevated CD41/CD42a
expression. PLPs predominantly exhibit an activated-like phenotype (CD62P+, degranulated
morphology), while microvesicles (100–500 nm) and exosomes (50–250 nm) displayed
size distributions and phenotypic markers consistent with native platelet-derived EVs. Proteomics
identified substantial core proteomes shared across fractions and fraction-specific
patterns consistent with selective cargo partitioning during EV biogenesis. Functional
enrichment indicated that MEG-01-derived vesicles preserve key hemostatic, cytoskeletal, and immune pathways commonly associated with platelet EV biology. Ingenuity Pathway
Analysis showed that PLPs exhibit proliferative transcriptional programs (elevated
MYC/RB1/TEAD1, reduced GATA1), while plasma exosomes display minimal differential
pathway activation compared to MEG-01 exosomes. Overall, these findings suggest that
MEG-01-derived EVs approximate certain aspects of megakaryocyte-lineage exosomes and
activated platelet-like states, although they do not fully replicate native platelet biology.
Notably, plasma exosomes show strong proteomic convergence with MEG-01 exosomes,
whereas platelet exosomes retain distinct activation-related features.},
organization = {FEDER/Junta de Andalucia  (B-CTS-676-UGR)},
organization = {Ministry of Economy and Competitiveness PID2023-152099OB-I00},
organization = {Fundación Roberto Arnall},
organization = {ROLUCAN Association (Rota Lucha contra el Cáncer)},
organization = {Research Trainees-Predoctoral student (PREDOC_01765)},
organization = {Spanish Health Institute Carlos III (FI17/00178)},
organization = {Ministry of Universities (FPU18/03410)},
organization = {University of Granada},
publisher = {MDPI},
keywords = {Meg-01 cell line},
keywords = {extracellular vesicles (EVs)},
keywords = {Platelets},
keywords = {Proteomics},
keywords = {Microvesicles (MVs)},
keywords = {Exosomes (EXOs)},
title = {Proteomic Validation of MEG-01-Derived Extracellular Vesicles as Representative Models for Megakaryocyte- and Platelet-Derived Extracellular Vesicles},
doi = {10.3390/biom15121698},
author = {Sánchez-Manas, José Manuel and Perales Romero, Sonia and Martínez Navajas, Gonzalo and Ceron-Hernández, Jorge and López Vázquez, Cristina María  and Peralbo-Molina, Ángela and Delgado, Juan Ramón and Martínez-Galán, Joaquina and Ramos-Mejía, Verónica and Chicano-Gálvez, Eduardo and Hernández-Valladares, María and Ortuño, Francisco Manuel and Torres Perales, Carolina and Real Luna, Pedro José},
}