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Proteomic profiling of extracellular vesicles suggests Collectin10 as potential biomarker in relapsing head and neck squamous cell carcinoma

ABSTRACT: Head and neck squamous cell carcinoma (HNSCC) is the 6th most common cancer worldwide. Developing new therapies has been ongoing for many decades, however, the 5-year overall survival rate remains comparably low and has not improved significantly. Treatment failure in HNSCC patients is common, especially in recurrences, and results in a poorer prognosis. Therefore, a better understanding of the disease is crucial to detect HNSCC recurrences at an early stage. HNSCC-associated extracellular vesicles (EVs), have been shown to suppress the immune system and thereby promote tumor progression. However, it is still unclear which pathways play a pivotal role in relapse or in remission. Contamination-free plasma EVs were purified using the EXÖBead® technology and EVs proteins underwent holistic proteome profiling by high-resolution mass spectrometry. COLEC10 appears to be the most regulated EVs protein and therefore has been chosen for further analysis and validation. Furthermore PanEV+ COLEC10+ and PanEV+ CD45Neg COLEC10+ of plasma EVs-EXÖBead complexes also showed a higher expression in relapsed patients compared to remission. Gene ontology analyses suggested that the relapse-upregulated proteins are significantly enriched in pathways for collagen trimer and chain formation, the complement system activation and in the tumor necrosis factor like domain superfamily related pathway. The remission-upregulated proteins were significantly enriched in early endosome or lysosomal membrane formation, membrane stability, mRNA stability and in ubiquitin protein ligase binding. According to common tissue RNA-seq database elevated COLEC10 gene has been associated with shorter survival in female patient and observed from different clinical cohort. Our results indicate that increased protein expression in EVs of patients with recurrent disease might contribute to disease progression with COLEC10 as potential biomarker in HNSCC.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Christina Ludwig

LAB HEAD: Christina Ludwig

PROVIDER: PXD033898 | Pride | 2022-11-09

REPOSITORIES: Pride

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			Action	DRS
	BBM_559_P146_02_FRH_001.raw	Raw
	BBM_559_P146_02_FRH_002.raw	Raw
	BBM_559_P146_02_FRH_004.raw	Raw
	BBM_559_P146_02_FRH_006.raw	Raw
	BBM_559_P146_02_FRH_008.raw	Raw

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Project description:IntroductionBacteria inhabit the in- and outside of the human body, such as skin, gut or the oral cavity where they play an innoxious, beneficial or even pathogenic role. It is well known that bacteria can secrete membrane vesicles (MVs) like eukaryotic cells with extracellular vesicles (EVs). Several studies indicate that bacterial membrane vesicles (bMVs) play a crucial role in microbiome-host interactions. However, the composition of such bMVs and their functionality under different culture conditions are still largely unknown. MethodsTo gain a better insight into bMVs, we investigated the composition and functionality of E. coli (DSM 105380) bMVs from the culture media Lysogeny broth (LB) and RPMI 1640 throughout the different phases of growth (lag-, log- and stationary-phase). bMVs from three time points (8 h, 54 h and 168 h) and two media (LB and RPMI 1640) were isolated by ultracentrifugation and analyzed using nanoparticle tracking analysis (NTA), cryogenic electron microscopy (Cryo-EM), conventional transmission electron microscopy (TEM) and mass spectrometry-based proteomics (LC-MS/MS). Furthermore, we examined pro-inflammatory cytokines IL-1β and IL-8 in the human monocyte cell line THP-1 upon bMV treatment.ResultsParticle numbers increased with inoculation periods. The bMV morphologies in Cryo-EM/TEM were similar at each time point and condition. Using proteomics, we identified 140 proteins, such as the common bMV markers OmpA and GroEL, present in bMVs isolated from both media and at all time points. Additionally, we were able to detect growth-condition-specific proteins. Treatment of THP-1 cells with bMVs of all six groups lead to significantly high IL-1β and IL-8 expressions.ConclusionsOur study showed that the choice of medium and the duration of culturing significantly influence both E.coli bMV numbers and protein composition. Our TEM/Cryo-EM results demonstrated the presence of intact E.coli bMVs. Common E. coli proteins, including OmpA, GroEL, and ribosome proteins, can consistently be identified across all six tested growth conditions. Furthermore, our functional assays imply that bMVs isolated from the six groups retain their function and result in comparable cytokine induction.

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Proteomic profiling of extracellular vesicles suggests Collectin10 as potential biomarker in relapsing head and neck squamous cell carcinoma

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