Neuroblastoma-derived small extracellular vesicles retaincellular identity and adrenergic/mesenchymal state signatures

Grimaldi, M.; Kirchner, M.; Szymansky, A.; Hendrix, A.; Muenster-Wandowski, A.; Gerhardt, H.; Hagemann, A. H.; Lodrini, M.; Auber, M.; Mertins, P.; Deubzer, H. E.

doi:10.64898/2026.09.21.753086

bioRxiv · 10.64898/2026.09.21.753086

Neuroblastoma-derived small extracellular vesicles retaincellular identity and adrenergic/mesenchymal state signatures

Grimaldi, M.·Kirchner, M.·Szymansky, A.·Hendrix, A.·Muenster-Wandowski, A.·Gerhardt, H.·Hagemann, A. H.·Lodrini, M.·Auber, M.·Mertins, P.·Deubzer, H. E.

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Abstract

Neuroblastoma is a heterogeneous pediatric malignancy characterized by distinct genomic subgroups and cell-state plasticity between adrenergic (ADRN) and mesenchymal (MES) phenotypes. While small extracellular vesicles (sEVs) mediate pre-metastatic niche priming, the degree to which sEV proteomes reflect parental genomic and phenotypic identity remains unresolved. Here, we isolated sEVs from six neuroblastoma cell models (IMR-5/75, BE(2)-C, GI-ME-N, CLB-GA, LAN-6, and SK-N-FI) representing three genomic subgroups (MYCN-amplified, TERT-rearranged, and unaltered) using OptiPrep density gradient centrifugation followed by size exclusion chromatography and ultrafiltration. High-resolution mass spectrometry of sEV proteomes revealed that sEVs carry highly reproducible protein signatures that tightly correlate with the parental genomic subgroups. Analyzing paired cell/sEV samples, we demonstrate cell-to-EV concordance of lineage-specific markers: ADRN sEVs selectively enriched L1CAM and dopamine {beta}-hydroxylase (DBH), whereas MES sEVs enriched SERPINE1 and type III collagen (COL3A1). These findings indicate that neuroblastoma-derived sEVs systematically preserve both the genomic landscape and the lineage transdifferentiation states of their cells of origin. This study establishes a robust molecular foundation for utilizing sEV-based proteomics as a surrogate to track cellular plasticity and for studying sEV-mediated microenvironmental remodeling.

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Grimaldi, M., Kirchner, M., Szymansky, A., Hendrix, A., Muenster-Wandowski, A., Gerhardt, H., Hagemann, A. H., Lodrini, M., Auber, M., Mertins, P., Deubzer, H. E.. 2026-09-22. Neuroblastoma-derived small extracellular vesicles retaincellular identity and adrenergic/mesenchymal state signatures. https://doi.org/10.64898/2026.09.21.753086

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