Renal cell carcinoma (RCC) is a lethal urogenital malignancy that accounts for approximately 2% of all malignancies and arises within a highly vascularized, metabolically active microenvironment. Mechanical forces, such as matrix stiffness and solid stress, promote tumor progression through mechano-transduction and metabolic reprogramming; however, the role of mechanical stress in regulating extracellular vesicle (EV)-mediated intercellular communication in RCC remains poorly understood. Here, we investigated whether mechanical stress alters the secretion, phenotype, and functional properties of tissue-derived EVs in RCC compared with benign renal oncocytoma (RO). Tissue specimens were obtained from 39 patients (31 with RCC and 8 with RO) to extract tissue-derived EVs before (Matrix-EVs) and after (Stress Matrix-EVs) the application of a mechanical stress. TEM confirmed the presence of intact EV-like particles within extract. NTA showed that mechanical stimulation increased EV yield. Surface-marker profiling was performed using a 37-marker panel by flow cytometry. Consistent CD9, CD63 and CD81 expression was observed in both Matrix- and Stress Matrix-EVs. In RCC samples, tissue-derived Stress Matrix-EVs were enriched with endothelial-associated markers (CD105, CD146), immune-associated markers (CD8, CD11c, CD86, CD69), and inflammatory markers (MCSP, ROR1) along with a decreased level of the epithelial marker CD326. These alterations were not observed in the comparison between Matrix- and Stress Matrix-EVs derived from RO tissues. Functional assays showed that RCC-derived Stress Matrix-EVs induced faster wound closure and higher degrees of cell migration in both normal HK-2 and malignant 786-O cells, along with the formation of immature angiogenic networks in HMEC-1 endothelial cells. In contrast, RO-derived Stress-Matrix EVs didn’t affect the migratory behavior of healthy and tumor cell lines, and caused different alterations of the angiogenetic network compared to RCC-derived EVs. Collectively, these findings reveal that mechanical stress selectively modulates EVs release from cell populations within malignant RCC tissues, promoting cell migration and angiogenesis. As such, the present study opens to new therapeutic possibilities to prevent or stop tumor progression.
Il carcinoma a cellule renali (RCC) è una neoplasia urogenitale letale che rappresenta circa il 2% di tutte le neoplasie e insorge in un microambiente altamente vascolarizzato e metabolicamente attivo. Le forze meccaniche, come la rigidità della matrice e lo stress solido, favoriscono la progressione tumorale attraverso la meccano-trasduzione e la riprogrammazione metabolica. Il ruolo dello stress meccanico nella regolazione della comunicazione intercellulare mediata da vescicole extracellulari (EV) nel RCC rimane ancora poco compreso. Questo studio ha come obiettivo quello di analizzare le alterazioni in secrezione, fenotipo e proprietà funzionali di EV tissutali in presenza di uno stress meccanico, in tessuti RCC e di oncocitoma renale benigno (RO). Campioni tissutali sono stati ottenuti da 39 pazienti (31 con RCC e 8 con RO) per estrarre EV derivate da tessuto prima (Matrix EVs) e dopo (Stress Matrix EVs) l’applicazione di uno stress meccanico. La microscopia elettronica a trasmissione (TEM) ha confermato la presenza di particelle integre simili a EV negli estratti. L’analisi NTA ha mostrato che la stimolazione meccanica aumenta la resa delle EV. Il profilo dei marcatori di superficie è stato valutato mediante citometria a flusso utilizzando un pannello di 37 marcatori. Matrix- e Stress Matrix-EVs hanno mostrato un’espressione comparabile di CD9, CD63 e CD81. Nei campioni di RCC, le Stress Matrix EVs risultavano arricchite in marcatori associati all’endotelio (CD105, CD146), marcatori immunitari (CD8, CD11c, CD86, CD69) e marcatori infiammatori (MCSP, ROR1), insieme a una riduzione del marcatore epiteliale CD326. Queste alterazioni non sono state osservate nel confronto tra Matrix e Stress Matrix EVs derivate da tessuti di RO. Saggi funzionali hanno evidenziato che le Stress Matrix EVs derivate da RCC inducono una chiusura della ferita (scratch) più rapida e un aumento della migrazione cellulare sia nelle cellule normali HK 2 sia nelle cellule tumorali 786 O, insieme alla formazione di reti angiogeniche immature nelle cellule endoteliali HMEC 1. Al contrario, le Stress Matrix EVs derivate da RO non influenzano il comportamento migratorio delle linee cellulari sane e tumorali e causano alterazioni diverse della rete angiogenica rispetto alle EV derivate da RCC. Nel complesso, questi risultati dimostrano che lo stress meccanico modula selettivamente il rilascio di EV da popolazioni cellulari all’interno dei tessuti maligni di RCC, promuovendo la migrazione cellulare e l’angiogenesi. Questo studio apre quindi nuove possibilità terapeutiche per prevenire o arrestare la progre.
Rilascio di vescicole extracellulari indotto da stress meccanico nel carcinoma a cellule renali: implicazioni per la crescita tumorale e la formazione di metastasi
DEYHIMI, ANAHITA SADAT
2025/2026
Abstract
Renal cell carcinoma (RCC) is a lethal urogenital malignancy that accounts for approximately 2% of all malignancies and arises within a highly vascularized, metabolically active microenvironment. Mechanical forces, such as matrix stiffness and solid stress, promote tumor progression through mechano-transduction and metabolic reprogramming; however, the role of mechanical stress in regulating extracellular vesicle (EV)-mediated intercellular communication in RCC remains poorly understood. Here, we investigated whether mechanical stress alters the secretion, phenotype, and functional properties of tissue-derived EVs in RCC compared with benign renal oncocytoma (RO). Tissue specimens were obtained from 39 patients (31 with RCC and 8 with RO) to extract tissue-derived EVs before (Matrix-EVs) and after (Stress Matrix-EVs) the application of a mechanical stress. TEM confirmed the presence of intact EV-like particles within extract. NTA showed that mechanical stimulation increased EV yield. Surface-marker profiling was performed using a 37-marker panel by flow cytometry. Consistent CD9, CD63 and CD81 expression was observed in both Matrix- and Stress Matrix-EVs. In RCC samples, tissue-derived Stress Matrix-EVs were enriched with endothelial-associated markers (CD105, CD146), immune-associated markers (CD8, CD11c, CD86, CD69), and inflammatory markers (MCSP, ROR1) along with a decreased level of the epithelial marker CD326. These alterations were not observed in the comparison between Matrix- and Stress Matrix-EVs derived from RO tissues. Functional assays showed that RCC-derived Stress Matrix-EVs induced faster wound closure and higher degrees of cell migration in both normal HK-2 and malignant 786-O cells, along with the formation of immature angiogenic networks in HMEC-1 endothelial cells. In contrast, RO-derived Stress-Matrix EVs didn’t affect the migratory behavior of healthy and tumor cell lines, and caused different alterations of the angiogenetic network compared to RCC-derived EVs. Collectively, these findings reveal that mechanical stress selectively modulates EVs release from cell populations within malignant RCC tissues, promoting cell migration and angiogenesis. As such, the present study opens to new therapeutic possibilities to prevent or stop tumor progression.| File | Dimensione | Formato | |
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Descrizione: Rilascio di vescicole extracellulari indotto da stress meccanico nel carcinoma a cellule renali: implicazioni per la crescita tumorale e la formazione di metastasi
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https://hdl.handle.net/20.500.14239/36241