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Saliva peptidome and proteome correlates with oral cancer prognosis

ABSTRACT: Protease activity has been associated with pathological processes that can lead to cancer development and progression. However, understanding the pathological unbalance in proteolysis is challenging since changes can occur simultaneously at protease, their inhibitor and substrate levels. Here, we present a pipeline that combines peptidomics, proteomics and peptidase predictions for studying proteolytic events in saliva associated with oral squamous cell carcinoma (OSCC) prognosis and lymph node metastasis. Our results suggest that cleavage products differentially abundant in the saliva of patients with (N+) or without (N0) nodal metastasis exhibit potential of prognostic value in oral cancer whereas reduced levels of peptidase inhibitors disturb the proteolytic balance in saliva of OSCC patients with worse prognosis.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Saliva

DISEASE(S): Oral Squamous Cell Carcinoma

SUBMITTER: Adriana Franco Paes Leme

LAB HEAD: Adriana Franco Paes Leme

PROVIDER: PXD020111 | Pride | 2020-11-13

REPOSITORIES: Pride

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			Action	DRS
	18fev_DaniGranato_N14.raw	Raw
	18fev_DaniGranato_N18.raw	Raw
	18fev_DaniGranato_N21.raw	Raw
	19fev_DaniGranato_N07.raw	Raw
	21Fev_DaniGranato_N22_CORRETO.raw	Raw

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Saliva peptidome correlates with oral cancer prognosis

Project description:Protease activity has been associated with pathological processes that can lead to cancer development and progression. However, understanding the pathological unbalance in proteolysis is challenging since changes can occur simultaneously at protease, their inhibitor and substrate levels. In this work we combined peptidomics, proteomics and peptidase predictions for studying proteolytic events in saliva associated with oral squamous cell carcinoma (OSCC) prognosis. Our results suggest that cleavage products differentially abundant in the saliva of patients with (N+) or without (N0) nodal metastasis exhibit potential of prognostic value in oral cancer and were are able to discriminate N+ and N0 patients whereas reduced protein levels of peptidase inhibitors might disturb the proteolytic balance in saliva of OSCC patients with poor prognosis

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Project description:Oral cavity cancer is one of the most common cancer worldwide and oral cavity squamous cell carcinoma (OSCC) accounts for up to 90 percent of oral cancers. In Taiwan, OSCC is the fifth most common malignancy of male and causes more than 250 deaths per year. The poor outcome of OSCC patients is principally ascribed to the fact that this disease is often advanced at the time of diagnosis, suggesting that early detection and prevention of OSCC are urgently needed to reduce the cancer burden. Analysis of cancer-related body fluids is one of promising approaches to identify cancer-related molecules and biomarker candidates. To identify salivary biomarkers of OSCC, salivary samples of OSCC patients, individuals with oral potentially malignant disorders (OPMD), and healthy volunteers were collected. With isobaric tags for relative and absolute quantitation (iTRAQ)-based mass spectrometry (MS) analysis, 1838 proteins were identified. The levels of 102 proteins are found to be elevated in the saliva samples of OSCC patients compared to that of OPMD and healthy groups. To improve the feasibility of biomarkers, we used a two-phase verification to measure these candidates. Among them, salivary levels of AHSG, CFH, FGA, and SERPINA1 were determined to be significantly higher in the OSCC patients than in the healthy or in the OPMD group with both of MRM-MS and ELISA. Furthermore, the levels of these proteins were highly related to tumor size, lymph-node metastasis, and cancer stages. The results of this study not only provides a powerful strategy for verification of biomarker candidates, but also suggested that AHSG, CFH, FGA, and SERPINA1 could act as salivary biomarkers for OSCC detection.

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Cell fusion between tumor cells and macrophages promotes the metastasis of OSCC patient through the activation of the chemokine signaling pathway

Project description:Tumor metastasis is responsible for the high mortality rate of patients with oral squamous cell carcinoma (OSCC). Although many hypotheses have been proposed to elucidate the mechanism of tumor metastasis, the origin of the metastatic tumor cells remains unclear. In this study, we explored the role of cell fusion in the formation of OSCC metastatic tumor cells. Murine OSCC tumor cells and macrophages were fused in vitro, and the cell proliferation, migration, and phagocytosis abilities of hybrid cells and parental cells were compared. Subsequently, we compared the transcriptome differences between hybrid and parental cells. Finally, we observed an association between the hybrid cells and the prognosis of patients with OSCC. Murine OSCC tumor cells and macrophages were successfully fused in vitro. The cytological and molecular experimental results revealed that OSCC tumor cells obtained a migration-related phenotype after fusion with macrophages, and the migration ability of hybrid cells was related to the activation of the “chemokine signal pathway”. Moreover, we proved that the existence of hybrid cells in OSCC tumor tissues is closely related to lymph node metastasis and poor prognosis. After fusion with macrophages, the chemokine signaling pathway in OSCC tumor cells was activated, leading to metastasis.

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The function and mechanism about how ORAOV1-B potentiates the OSCC metastasis.

Project description:Lymph node metastasis, a powerful prognostic indicator of oral squamous cell carcinoma (OSCC), is chiefly responsible for cancer death. Long non-coding RNA (lncRNA) is recently addressed to significantly account for modulating OSCC metastasis. Here, we identified a novel alternative splice of Oral Cancer Overexpressed 1 (ORAOV1), ORAOV1-B, which was subsequently validated as an lncRNA and correlated with OSCC lymph node metastasis; significantly increased invasion and migration were observed in ORAOV1-B-overexpressed OSCC cells; In the metastatic model, ORAOV1-B also significantly contributed to OSCC-related lung metastasis. cDNA microarrays was performed to compare up- or down-regulated genes in EV and ORAOV1-B sets of OSCC, which suggested TNFα as a potential downstream of ORAOV1-B and the upregulation of pro-inflammatory genes, indicating the activation of NF-κB pathway. In summary, the novel splice variant ORAOV1-B is an lncRNA, which significantly potentiates OSCC invasion and metastasis by binding to Hsp90 and activating the NF-κB-TNFα loop. Our study implies that ORAOV1-B might serve as an attractive OSCC metastasis intervention target.

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Project description:Oral squamous cell carcinoma (OSCC) has increased morbidity, and its high malignant potential, including metastasis, affects patient survival. Bromodomain containing 4 (BRD4) is a chromatin protein that recognizes and associates with acetylated histone lysines, and facilitates transcriptional activation. BRD4 has been implicated in cell proliferation, metastasis, and prognosis in several types of cancer. However, the role of BRD4 in OSCC remains to be elucidated. Herein, we investigated the role of BRD4 and the potential utility of BRD4 as a therapeutic target in OSCC. JQ1, which inhibits the interaction between BRD4 and acetylated histones, suppressed the proliferation, migration, and invasion of OSCC cell lines. JQ1 reduced the expression levels of 15 metastasis genes in OSCC, including the Matrix Metallopeptidase 2 (MMP2) gene. Interestingly, our chromatin immunoprecipitation assay showed that JQ1 reduced the BRD4 binding to the histone H3 lysine 27 acetylation-enriched sites in the MMP2 locus. Finally, qRT-PCR analyses of biopsy specimens from OSCC patients revealed that the BRD4 and MMP2 expression levels were correlated in the cancerous regions, and both were highly expressed in lymph node metastasis cases, including delayed metastasis. These results suggest that BRD4 contributes to metastasis in OSCC, through the epigenetic regulation of the MMP2 gene, and thus BRD4 may represent a therapeutic target and a novel prediction indicator for metastasis.

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