4.6 Article

Phosphoproteome analysis of cerebrospinal fluid extracellular vesicles in primary central nervous system lymphoma

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ANALYST
卷 148, 期 15, 页码 3594-3602

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ROYAL SOC CHEMISTRY
DOI: 10.1039/d3an00670k

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Primary central nervous system lymphoma (PCNSL) is a rare and aggressive type of non-Hodgkin's lymphoma, posing challenges in diagnosis and treatment. This study introduces a method called EVTRAP for efficient isolation of extracellular vesicles (EVs) from cerebrospinal fluid (CSF) and applies it to analyze phosphoproteomics in PCNSL patients. The results demonstrate the feasibility of EVTRAP-based strategy in detecting molecular markers for PCNSL.
Primary central nervous system lymphoma (PCNSL) is a rare but highly aggressive extra-nodal non-Hodgkin's lymphoma, mostly of the diffuse large B-cell lymphoma (DLBCL) type. The present invasive diagnosis and poor prognosis of PCNSL propose an urgent need to develop molecular markers for early detection, real-time monitoring and treatment evaluation. Cerebrospinal fluid (CSF)-derived extracellular vesicles (EVs) are promising biomarker carriers for liquid biopsy of CNS diseases and brain tumors; however, research remains challenging due to the low concentration of EVs in the limited available volume of CSF from each individual patient and the low efficiency of existing methods for EV enrichment. Here, we introduce functionalized magnetic beads called EVTRAP (extracellular vesicles total recovery and purification) for rapid and efficient EV isolation from CSF. By coupling with high-performance mass spectrometry, over 19 000 peptides representing 1841 proteins were identified from just 30 & mu;L of CSF. Furthermore, up to 3000 phosphopeptides representing over 1000 phosphoproteins were identified from about 2 mL of CSF. Finally, we analyzed the EV phosphoproteomics of CSF samples from PCNSL patients and non-PCNSL controls. Among them, multiple phosphoproteins related to PCNSL, including SPP1, MARCKS, NPM1 and VIM, were shown to be up-regulated in the PCNSL group. These results demonstrated the feasibility of the EVTRAP-based analytical strategy in CSF EV phosphoproteomic analysis of PCNSL molecular markers.

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