The RNA cargo of plasma-derived extracellular vesicles in mCRPC patients captures cancer cells and tumor microenvironment signals

Vannuccini, F.; Ciani, Y.; Mugoni, V.; Quaini, O.; Orlando, F.; Weber, V.; Martinez, A. C.; Martinelli, A.; Biada, E.; Marinelli, A.; Casanova-Salas, I.; Pessina, F.; Di Vizio, D.; Vannini, A.; Cremaschi, P.; Lise, S.; Nardella, C.; De Giorgi, U.; Buttigliero, C.; Caffo, O.; Basso, U.; Mateo, J.; Attard, G.; Demichelis, F.

doi:10.64898/2026.09.03.747025

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bioRxiv · 10.64898/2026.09.03.747025

The RNA cargo of plasma-derived extracellular vesicles in mCRPC patients captures cancer cells and tumor microenvironment signals

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Abstract

Circulating analytes in cancer patients capture tumor-related signals. We profiled matched extracellular vesicle (EV) total RNA and cell-free DNA (cfDNA) from the same plasma aliquots of chemo-naive metastatic castration-resistant prostate cancer (mCRPC) patients treated with Enzalutamide (n=54 patients, n=119 longitudinal samples; NCT06981377) at four Italian clinical centers and interrogated data from >10,000 cancer patients' and healthy individuals' samples. Transcript integrity analysis identified coding and non-coding species with fragmentation patterns varying across RNA biotypes. EV-RNA data deconvolution revealed signal from immune populations, including fractions classified as CD4+ T cells, whose abundance increased with disease progression in plasma EVs and mCRPC tissues. By leveraging tissue data-informed mining, we established a novel prostate cancer-related EV-RNA signature that resulted in an independent predictor of poor prognosis and captured tumor microenvironment-derived signals. Integrating EV-RNA and ctDNA information improved patient stratification for progression-free survival. These findings suggest a multifaceted role for plasma EVs as a source of cancer biomarkers.

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Vannuccini, F., Ciani, Y., Mugoni, V., Quaini, O., Orlando, F., Weber, V., Martinez, A. C., Martinelli, A., Biada, E., Marinelli, A., Casanova-Salas, I., Pessina, F., Di Vizio, D., Vannini, A., Cremaschi, P., Lise, S., Nardella, C., De Giorgi, U., Buttigliero, C., Caffo, O., Basso, U., Mateo, J., Attard, G., Demichelis, F.. 2026-09-07. The RNA cargo of plasma-derived extracellular vesicles in mCRPC patients captures cancer cells and tumor microenvironment signals. https://doi.org/10.64898/2026.09.03.747025

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