Functional and Proteomic Profiles of CD3(+) Plasma‐Derived Small Extracellular Vesicles Differentiate Cancer Patients From Healthy Donors

Abstract

Small extracellular vesicles (sEV) released by T cells play a key role in immune regulation. Immune capture with anti‐CD3 antibodies was used to isolate and study T cell‐derived CD3(+)sEV from the plasma of patients with melanoma (MPs) or healthy donors (HDs). Functional responses induced in recipient target cells by CD3(+)sEV of MPs differed from responses induced by CD3(+)sEV of HDs. Approximating functions mediated by melanoma cell‐derived sEV (MTEX), CD3(+)sEV of MPs reduced metabolic activity and proliferation of T cells while promoting activity in Mel526 cell targets. Proteomics profiling confirmed functional differences between CD3(+)sEV of MPs and HDs. Of 294 sEV‐specific proteins identified in CD3(+)sEV, 226 were detected in the parent T cell proteome, confirming that the CD3(+)sEV proteome mimics that of the parent T lymphocytes. Among them were 66 differentially expressed proteins (DEPs) that differentiated vesicles from MPs and HDs. These DEPs were associated with processes linked to cancer‐related functions. DEPs upregulated in CD3(+)sEV of MPs were associated with RHO‐GTPase, cytokine, and MAPK signaling pathways. Thus, T cells of MPs were reprogrammed by MTEX to produce CD3(+)sEV that functionally resembled MTEX, partly recapitulated features of the tumor proteome, and differed from CD3(+)sEV of HDs. In cancer, the TEX‐rewired T cells produce CD3(+)sEV that potentially could serve as a liquid biopsy of patients’ T cells.