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Proteomic analysis of serum small extracellular vesicles identifies diagnostic biomarkers for neuroblastoma

ABSTRACT: The search for new biomarkers for neuroblastoma diagnosis is driven by the need for improved accuracy, early detection, and personalized treatment of this particular type of cancer. Neuroblastoma is a pediatric cancer that primarily affects young children, and its diagnosis can be challenging due to its diverse clinical presentation and variable outcomes. Small extracellular vesicles (sEVs) have gained attention as potential tumor biomarkers due to their unique characteristics and their ability to reflect the molecular profile of the parent tumor cells. sEVs released by tumor cells can carry a cargo of proteins, nucleic acids (such as DNA and RNA), and other molecules that reflect the molecular diversity of the tumor. By analyzing the content of sEVs, researchers can gain insights into the heterogeneity of the tumor and potentially identify specific biomarkers associated with tumor subtypes or disease progression. sEVs can be found in various body fluids, this accessibility makes sEVs a promising source for non-invasive or minimally invasive diagnostic and monitoring approaches. sEVs are protected by a lipid bilayer membrane, which shields their cargo from degradation by enzymes and other harsh conditions in the extracellular environment. This stability allows sEVs to circulate in body fluids for extended periods, making them suitable for biomarker analysis. Here, we performed mass spectrometry-based proteomic analysis of serum sEVs to identify and validate potential protein biomarkers for diagnosis of NB.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Serum

SUBMITTER: Dongrui Ji

LAB HEAD: Lin Yang

PROVIDER: PXD048511 | Pride | 2024-01-13

REPOSITORIES: Pride

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Project description:Saliva, a biofluid enriched in biological omic constituents, has emerged as a promising source for exosomal biomarkers due to its easy accessibility. Despite the understanding of the coronavirus disease-19 (COVID-19), the role of Salivary Extracellular Vesicles (sEVs) in COVID-19 remains poorly understood. Exploring the proteomic cargo of sEVs could prove valuable for diagnostic and prognostic purposes in assessing COVID-19. The proteomic cargo of sEVs from COVID-19 (+) subjects and their healthy close contacts (HCC) was explored. Nine COVID-19 positive (+) patients and eleven in-house close contact patients identified by real-time quantitative polymerase chain reaction (RT-qPCR) of nasopharyngeal swabs were included. In-house close contacts were defined as individuals with a negative RT-qPCR result sharing a residence with a confirmed COVID-19 case. sEVs were isolated by ultracentrifugation from unstimulated saliva samples, and subsequently characterized through nanoparticle tracking, transmission electron microscopy, and western-blot analyses. The proteomic cargo of sEVs was processed by LC-MS/MS. sEVs were morphologically compatible with EVs, with the presence of Syntenin-1 and CD81 EVs markers. The sEVs proteome showed 1,417 proteins: 1,288 in COVID-19 (+) cases and 1,382 in HCC. 35 proteins were found exclusively and 89 were more abundant in sEVs from COVID-19 (+) subjects. “Coronavirus disease response”, “complement and coagulation cascades”, and “PMN extracellular trap formation” were the most enriched KEGG pathways in COVID-19 (+) cases. The most represented biological processes were “Hemoglobin and haptoglobin binding” and “oxygen carrier activity”, and the best-denoted molecular functions were “regulated exocytosis and secretion” and “leucocyte and PMN mediated immunity”. We suggest that sEVs proteomic cargo in COVID-19 is related to immune response processes, oxygen transport, and antioxidant mechanisms. In contrast, in HCC, sEVs signature profiles are mainly associated with epithelial homeostasis.

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Proteomic analysis of serum small extracellular vesicles identifies diagnostic biomarkers for neuroblastoma

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