CD19.CAR T-cell–derived extracellular vesicles express CAR and kill leukemic cells, contributing to antineoplastic therapy

Chimeric antigen receptor (CAR) T-cell-derived extracellular vesicles (EVs) might represent a new therapeutic tool for boosting CAR T-cell antileukemic effects. Here, a cohort of 22 patients who received infusion with CD19 CAR T cells were monitored for the presence of circulating CD19 CAR+ T-cell-derived EVs (CD19.CAR+EVs), which were then separated and functionally characterized for their killing abilities. A good manufacturing practice (GMP)-compliant separation method was also developed. Results demonstrated that CD19.CAR+EVs were detectable in peripheral blood up to 2 years after infusion, indicating long-lasting persistence of their parental cells. Notably, early decreases of circulating CD19.CAR+EV concentrations correlated with failure of CAR T-cell therapy. Circulating CD19.CAR+EVs displayed a median size (standard deviation) of 133.1 ± 65.5 nm and carried a proapoptotic protein cargo. These EVs expressed higher CAR levels than their parental cells. Furthermore, CD19.CAR+EVs did not activate heterologous T cells and produced significant, specific, and dose-dependent cytotoxic effects on CD19+ cell lines and primary cells. The new GMP-compliant EV isolation method allowed for a recovery of 63% ± 5.7% of CD19.CAR+EVs. A deeper analysis of the different protein cargoes carried by EVs derived from different CAR T-cell subpopulations identified a proapoptotic functional pathway linked to CD8+LAG-3+ EVs. Overall, our data indicate that CD19.CAR+EVs may be proposed as promising dynamic new biomarkers of CAR T-cell activity and, by contributing to the direct killing of leukemic targets, represent a new product with strong therapeutic potential that could be infused independently of CAR T cells.

Authors

Institutions

Publication Details

Journal
Blood Advances
Published
2025-06-11
DOI
https://doi.org/10.1182/bloodadvances.2024014860
Citations
14
Primary Topic
CAR-T cell therapy research
Type
article
Field-Weighted Citation Impact
14.64

Funders

Controls
|||
ALL TIME
JAN
FEB
MAR
APR
MAY
JUN
JUL
AUG
SEP
OCT
article

CD19.CAR T-cell–derived extracellular vesicles express CAR and kill leukemic cells, contributing to antineoplastic therapy

Damiana Pieragostino, Elsa Pennese, Paola Lanuti, Anna Piro et al.
14 citations
Blood Advances
CAR-T cell therapy research
14.64
article

CD19.CAR T-cell–derived extracellular vesicles express CAR and kill leukemic cells, contributing to antineoplastic therapy

Damiana Pieragostino, Elsa Pennese, Paola Lanuti, Anna Piro, Antonella Fontana, Bianca Fabi, Ida Villanova, Laura De Lellis, Barbara Di Francesco, Nicola Tinari, Francesco Restuccia, Piero Del Boccio, Pasquale Simeone, Stefano Baldoni, Davide Brocco, Giulia Corradi, Alessandro Cama, Maria Vittoria Mattoli, Francesco Guardalupi, Mauro Di Ianni, Raffaella Giancola, Rosalba Florio, Domenico De Bellis, Serena Veschi, Maria Concetta Cufaro, Prassede Salutari, Fabio Verginelli, Fabrizia Colasante, Serena Pilato, Stella Santarone, Simone Vespa, Francesca D’Ascanio, Lucia Di Re, Annalisa Natale, Isabella D’Amario
article en
14 citations

Abstract

Chimeric antigen receptor (CAR) T-cell-derived extracellular vesicles (EVs) might represent a new therapeutic tool for boosting CAR T-cell antileukemic effects. Here, a cohort of 22 patients who received infusion with CD19 CAR T cells were monitored for the presence of circulating CD19 CAR+ T-cell-derived EVs (CD19.CAR+EVs), which were then separated and functionally characterized for their killing abilities. A good manufacturing practice (GMP)-compliant separation method was also developed. Results demonstrated that CD19.CAR+EVs were detectable in peripheral blood up to 2 years after infusion, indicating long-lasting persistence of their parental cells. Notably, early decreases of circulating CD19.CAR+EV concentrations correlated with failure of CAR T-cell therapy. Circulating CD19.CAR+EVs displayed a median size (standard deviation) of 133.1 ± 65.5 nm and carried a proapoptotic protein cargo. These EVs expressed higher CAR levels than their parental cells. Furthermore, CD19.CAR+EVs did not activate heterologous T cells and produced significant, specific, and dose-dependent cytotoxic effects on CD19+ cell lines and primary cells. The new GMP-compliant EV isolation method allowed for a recovery of 63% ± 5.7% of CD19.CAR+EVs. A deeper analysis of the different protein cargoes carried by EVs derived from different CAR T-cell subpopulations identified a proapoptotic functional pathway linked to CD8+LAG-3+ EVs. Overall, our data indicate that CD19.CAR+EVs may be proposed as promising dynamic new biomarkers of CAR T-cell activity and, by contributing to the direct killing of leukemic targets, represent a new product with strong therapeutic potential that could be infused independently of CAR T cells.

Blood AdvancesVol. 9(12)
University of Chieti-Pescara (IT), Azienda USL di Pescara (IT)
European Commission, Associazione Italiana Contro le Leucemie-Linfomi e Mieloma, European Regional Development Fund, European Social Fund
Openalex Percentile: Top 1%
CAR-T cell therapy research
14.64
DOIOpen Access PDFOpenAlex
AI Navigator

Ask Laika to Summarize, Analyze, and Connect papers live on the map.

Summarize Papers & Methodologies

Extract key findings, datasets, and comparative methods across publications.

Benchmark Rankings & Visual Analytics

Rank top research institutions, authors, funders, topics, and journals by Field-Weighted Citation Impact (FWCI) and paper volume with instant charts.

Connect Distant Disciplines

Bridge topological clusters on the map to find hidden collaborative intersections.