Exosomes from Osteosarcoma and normal osteoblast differ in proteomic cargo and immunomodulatory effects on T cells

Troyer, Ryan M.; Ruby, Carl E.; Goodall, Cheri P.; Yang, Liping; Maier, Christoph; Albarqi, Hassan A.; Brady, Jacqueline V.; Bathke, Kallan; Taratula, Oleh; Mourich, Dan V.; Bracha, Shay

doi:10.1016/j.yexcr.2017.07.011

Cited by 64 publications

(52 citation statements)

References 30 publications

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“…Circulating exosomes of five dogs were used for validation of exosome isolation and measured using NanoSight light scatter technology ( Figure 1 ). The measured vesicles' size mean was in a range of 113.2-173.3 and a mode of 90.1-166.9, which is consistent with the size of exosomes measured in previous studies using similar technology [5] , [14] .…”

Section: Resultssupporting

confidence: 89%

“…The analysis of exosomes from cell culture supernatants was done on samples which were diluted to a concentration of 50-150 particles per frame. The chamber conditions and system settings were adjusted as previously described [14] .…”

Section: Methodsmentioning

confidence: 99%

“…The roles of exosomes in the disease course of osteosarcoma or response to therapeutic treatments have not been fully characterized [13] , [14] . Exosomal proteins derived from human osteosarcoma cell line supernatants are enriched with proteins associated with angiogenesis, cell adhesion, immune evasion, and cell migration in comparison to proteins in exosomes from nonmalignant cells and non–exosome-derived blood proteins [13] , [14] , [15] . Recent investigations in human pediatric OSA have demonstrated significant differences in serum protein content; proteins which differ can be used as biomarkers for early detection of cancer, metastasis, and response to therapy [16] .…”

Section: Introductionmentioning

confidence: 99%

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A Preliminary Proteomic Investigation of Circulating Exosomes and Discovery of Biomarkers Associated with the Progression of Osteosarcoma in a Clinical Model of Spontaneous Disease

Brady

Troyer

Ramsey

et al. 2018

Translational Oncology

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Circulating cancer exosomes are microvesicles which originate from malignant cells and other organs influenced by the disease and can be found in blood. The exosomal proteomic cargo can often be traced to the cells from which they originated, reflecting the physiological status of these cells. The similarities between cancer exosomes and the tumor cells they originate from exhibit the potential of these vesicles as an invaluable target for liquid biopsies. Exosomes were isolated from the serum of eight osteosarcoma-bearing dogs, five healthy dogs, and five dogs with traumatic fractures. We also characterized exosomes which were collected longitudinally from patients with osteosarcoma prior and 2 weeks after amputation, and eventually upon detection of lung metastasis. Exosomal proteins fraction were analyzed by label-free mass spectrometry proteomics and were validated with immunoblots of selected proteins. Ten exosomal proteins were found that collectively discriminate serum of osteosarcoma patients from serum healthy or fractured dogs with an accuracy of 85%. Additionally, serum from different disease stages could be distinguished with an accuracy of 77% based on exosomal proteomic composition. The most discriminating protein changes for both sample group comparisons were related to complement regulation, suggesting an immune evasion mechanism in early stages of osteosarcoma as well as in advanced disease.

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Section: Resultssupporting

confidence: 89%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Preliminary Proteomic Investigation of Circulating Exosomes and Discovery of Biomarkers Associated with the Progression of Osteosarcoma in a Clinical Model of Spontaneous Disease

Brady

Troyer

Ramsey

et al. 2018

Translational Oncology

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“…We selected 31 hypoxia-responsive protein targets (Table 1 and Supplemental Table S4) relevant to cancer biology belonging to the following pathways: glycolysis and pentose phosphate pathway (PPP), collagen biosynthesis, reactive oxygen species (ROS) generation/redox regulation, and immunomodulation. The reason for selecting proteins that have functions associated with immune responses was based on our recent work showing that OS-derived exosomes contained higher amounts of immunomodulatory cargo compared with normal osteoblasts 24 . In addition, we added HIF1A and LOX as additional targets due to their relevance in cancer aggressiveness and adaption of ECM to promote invasion.…”

Section: Resultsmentioning

confidence: 99%

Delineation of hypoxia-induced proteome shifts in osteosarcoma cells with different metastatic propensities

Song

Pearce

Jiang

et al. 2020

Sci Rep

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osteosarcoma (oS) is the most common bone cancer in children and young adults. Solid tumors are characterized by intratumoral hypoxia, and hypoxic cells are associated with the transformation to aggressive phenotype and metastasis. the proteome needed to support an aggressive osteosarcoma cell phenotype remains largely undefined. To link metastatic propensity to a hypoxia-induced proteotype, we compared the protein profiles of two isogenic canine OS cell lines, POS (low metastatic) and HMpoS (highly metastatic), under normoxia and hypoxia. Label-free shotgun proteomics was applied to comprehensively characterize the hypoxia-responsive proteome profiles in the OS cell phenotypes. Hypothesis-driven parallel reaction monitoring was used to validate the differential proteins observed in the shotgun data and to monitor proteins of which we expected to exhibit hypoxia responsiveness, but which were absent in the label-free shotgun data. We established a "distance" score (|z HMPOS − z POS |), and "sensitivity" score (|z Hypoxia − z Normoxia ) to quantitatively evaluate the proteome shifts exhibited by oS cells in response to hypoxia. evaluation of the sensitivity scores for the proteome shifts observed and principal component analysis of the hypoxia-responsive proteins indicated that both cell types acquire a proteome that supports a Warburg phenotype with enhanced cell migration and proliferation characteristics. Cell migration and glucose uptake assays combined with protein function inhibitor studies provided further support that hypoxia-driven adaption of pathways associated with glycolytic metabolism, collagen biosynthesis and remodeling, redox regulation and immunomodulatory proteins typify a proteotype associated with an aggressive cancer cell phenotype. Our findings further suggest that proteins involved in collagen remodeling and immune editing may warrant further evaluation as potential targets for anti-metastatic treatment strategies in osteosarcoma.

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“…B16-F10 exosomes also decreased the proportion of proliferating PD-1 + CD8 T cells in both spleen and lymph nodes, suggesting that exosomal PD-L1 suppresses anti-tumour immunity systemically [24]. Troyer et al reported the negative effect of OS exosomes on CD4 + and CD8 + T cell proliferation was much greater than that of osteoblast derived exosomes [41]. In the present study, we showed that in the metastatic model, mice injected with143B-Rab27a-KD, which failed to secrete more exosomes, had abundant metastatic lesions in the lung compared to ones received injection of wild type 143B osteosarcoma cell.…”

Section: Discussionmentioning

confidence: 99%

Exosomal PD-L1 Derived From Osteosarcoma is Associated With Pulmonary Metastasis for Patients With Osteosarcoma

Wang¹,

Zhang²,

Sun³

et al. 2020

Preprint

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Abstract Background Recent studies indicated that exosomal programmed death-ligand 1 (PD-L1) derived from cancers could induce immunosuppression and tumor pathogenesis. However, it is unclear how the exosome influences the osteosarcoma (OS) progression and whether PD-L1 also exists in serum exosomes (Sr-exosomes) of patients with osteosarcoma. Methods We examined serum exosomes from 70 patients with osteosarcoma, 9 patients with benign tumors and 22 healthy donors. Osteosarcoma-derived exosomes and exosomal-PD-L1 were functionally evaluated using in vivo and in vitro models. Results The characteristics of serum exosomes of OS patients and OS cell line exosomes were confirmed by several methods. Bioinformatic analysis demonstrated that Sr-exosomes isolated from OS patients may involve in the important process of immune function and cancer pathogenesis for OS patients. Meanwhile, exosomes derived from OS cell line and serum of OS patients could induce osteosarcoma migration and invision in vitro. Then, we confirmed PD-L1 existing in Sr-exosomes of patients with osteosarcoma and higher level of Sr-exosomal PD-L1 in OS patients compared to healthy donors and patients with benign tumor. Furthermore, exosomal-PD-L1 derived from osteosarcoma promoted the pulmonary metastasis, which was demonstrated in the osteosarcoma metastatic model. Conclusion We confirm that osteosarcoma releases the exosomes carrying PD-L1 on their surface and induces the pathogenesis of pulmonary metastasis for OS patients.

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Exosomes from Osteosarcoma and normal osteoblast differ in proteomic cargo and immunomodulatory effects on T cells

Cited by 64 publications

References 30 publications

A Preliminary Proteomic Investigation of Circulating Exosomes and Discovery of Biomarkers Associated with the Progression of Osteosarcoma in a Clinical Model of Spontaneous Disease

A Preliminary Proteomic Investigation of Circulating Exosomes and Discovery of Biomarkers Associated with the Progression of Osteosarcoma in a Clinical Model of Spontaneous Disease

Delineation of hypoxia-induced proteome shifts in osteosarcoma cells with different metastatic propensities

Exosomal PD-L1 Derived From Osteosarcoma is Associated With Pulmonary Metastasis for Patients With Osteosarcoma

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