Osteosarcoma‐derived extracellular vesicles induce a tumor‐like phenotype in normal recipient cells

Abstract

Osteosarcoma is the most common primary bone cancer and the most frequent cause of bone cancer-related deaths in children and adolescents. Osteosarcoma cells are able to establish a crosstalk with resident bone cells leading to the formation of a deleterious vicious cycle. We hypothesized that osteosarcoma cells can release, in the bone microenvironment, transforming Extracellular Vesicles (EVs) involved in regulating bone cell proliferation and differentiation, thereby promoting tumor growth. We assessed EV production by three osteosarcoma cell lines with increasing aggressiveness in order to investigate their roles in the communication between osteosarcoma cells and normal recipient cells. Osteosarcoma-derived EVs were used to treat the murine fibroblast cell line NIH3T3 and to study the induction of tumor-like phenotypes. Our results showed that osteosarcoma cell lines are able to produce EVs that fuse to recipient cells, with a very high uptake efficiency. The treatment of recipient NIH3T3 with osteosarcoma-derived EVs induced substantial biological and functional effects, as an enhanced proliferation and survival capability under starved conditions, high levels of activated survival pathways, an increased migration, adhesion, and 3D sphere formation and the acquired capability to grow in an anchorage-independent manner. Moreover, in murine NIH3T3 we found human mRNAs of TNF-α, IL-6, and TGF-β, as well as a de novo expression of murine MMP-9 and TNF-α following the treatment of human osteosarcoma-derived EVs.

Epigenetic alterations in mesenchymal stem cells by osteosarcoma-derived extracellular vesicles

“…EVs are therefore an excellent tool for studying the interaction between the tumour and its surrounding cells. Cross-talk between MSCs and OS has demonstrated the oncogenic potential of OS-EVs in recipient cells [37,41,42]. For example, in an in vivo mouse study, OS-EVs carrying functional TGFβ (transforming growth factor β) were internalized by MSCs and altered the MSCs phenotype towards a pro-tumorigenic and pro-metastatic phenotype by activating oncogenic IL&-STAT3 signalling pathways, thereby promoting tumour growth and metastasis [21].…”

“…Global LINE-1 hypomethylation contribute to cancer development by inducing genetic instability, and predisposing the cells to chromosomal defects, transcription disruption, insertion mutations, and thus influencing the overall gene expression [50][51][52]. OS-EV treated cells have been shown to acquire tumour-phenotype characteristics such as increased adhesion, proliferation, migration rate, and anchorage-independent growth [42]. Subsequently, the effects of OS-EVs on recipient cells were investigated by focusing on analyzing the expression of genes (MMP1, VEGF-A, ICAM1) related only to bone microenvironment remodelling.…”

Extracellular Vesicles in Bladder Cancer: Biomarkers and Beyond

“…The transformation of healthy cells into malignant cancer cells involves several pathologic processes and many studies indicate that TEVs participate by transferring oncogenic cargo molecules to recipient cells [ 6 ]. A study by Urciuoli et al [ 7 ] reported that treating NIH3T3 fibroblasts with osteosarcoma-derived EVs induced tumor-like phenotypes. Cells gained survival capacity by enhanced proliferation, migration, adhesion and 3D sphere formation and acquired the ability to grow in an anchorage dependent manner.…”

Proteomic Analysis of Extracellular Vesicles for Cancer Diagnostics

“…Surface LPDs [120] HTL/ BCL Exo UC MICA/B, ULBP1/2 Surface LPDs [121] OS EV UC, ExoQuick TNF-α, IL-6, TGF-β Inside Malignant neoplasm [122] OS EV UC, ExoQuick NPM1, CCT2, CCT4, CCT6A, CCT8, VIM, CLTC, COL6A2, HNRNPC, PKM, ACTN4, MYH10, PAICS, VCP, ANXA1, ACLY N/A Malignant neoplasm, Metastasis [123] Brain Glioma EV UC PTRF/Cavin1 N/A Tumor growth [124] GSC EV UC VEGF-A N/A Angiogenesis [125] GBM EV UC, OptiPrep Annexin A1, ACTR3, integrin β1, IGF2R, PDCD6IP N/A N/A [126] GBM MV UC Angiogenin, FGFα, IL-6, IL-8, TIMP-1, VEGF, TIMP-2 N/A N/A [127] Breast Breast cancer cell EV UC TrpC5 N/A Tumor drug-resistance [128] Breast cancer cell MV UC TF N/A Aggressive tumor growth [129] Breast cancer cell MV, Exo UC EMMPRIN, Wnt 5a N/A Malignant invasion [130] Colon CRC Exo UC, DG,IAC FAT1, CDH17, CEACAM1, CEACAM5 N/A N/A [108] Primary CRC, Metastatic CRC Exo UC, OptiPrep MET, TNIK N/A Tumor progression [131] Colon cancer cell Exo UF, OptiPrep, IAC A33, cadherin-17, CEP55, CLDN1/3/7, EGFR, EPHA2, KRT18, mucin 13, PCNA, POLD1, PPP2R1B, CEA, EpCAM, MLK-1, MAPK-4, Ephrin-B1, Ephrin-B2, RUVBL1 N/A N/A [132] Kidney CCRCC EV UC AZU1 Surface Membrane permeabilizing activity [133] RCC Exo UC, OptiPrep CP, MMP9, PODXL, CAIX, DKK4 N/A N/A [134] Liver HCC Exo UC 14-3-3ζ N/A T cells exhaustion [135] HCC cell lines Exo UC HGF N/A Sorafenib resistance [136] Lung NSCLC Exo Micro array (IAC) NY-ESO-1, EGFR, PLAP, EpCam Surface N/A [137] NSCLC Exo UC LRG1 N/A N/A [138] Lung cancer (ADC, SCC)…”