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Serum Exosome Proteome of Tuberculosis Patients Demonstrated Enriched Inflammatory Response Proteins

ABSTRACT: Detailed understanding of host pathogen interaction in tuberculosis is an important avenue for identifying novel therapeutic targets. Extracellular vesicles like exosomes that are rich in protein, nucleic acids and lipids, act as messenger and may show altered composition in infection. In this case control study, we isolated exosomes from serum of 52 subjects (all male, 33 (17-70) in years) including drug naïve active tuberculosis (ATB: n=20), non-tuberculosis (NTB: n=16) and healthy subjects (n=16). Serum exosomal proteome analysis was carried out using isobaric tag for relative and absolute quantification (iTRAQ) experiments and an independent test sample sets (n=36) was used for validation. A set of 132 and 68 proteins were identified in iTRAQ-I (ATB/Healthy) and iTRAQ-II (ATB/NTB) experiments respectively. Analysis of data identified a set of 4 important proteins (KYAT3, SERPINA1, HP and APOC3) that could differentiate ATB subjects from healthy controls successfully and Western data corroborated iTRAQ findings. Identification of such a protein panel in circulating exosomes besides providing novel insights into their role in tuberculosis may prove to be useful targets to develop novel host-directed therapeutic intervention.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Dr. Ranjan Kumar Nanda

PROVIDER: MSV000081833 | MassIVE | Tue Dec 19 22:47:00 GMT 2017

REPOSITORIES: MassIVE

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Project description:Clinical management of metastatic gastric cancer (mGC) remains a major challenge due to a lack of specific biomarkers and effective therapeutic targets. Recently, accumulating evidence has suggested that exosomes play an essential role in cancer metastasis and can be an excellent reservoir of novel biomarkers and candidate therapeutic targets for cancer. Therefore, in this study, we aimed to reveal the proteomic profile of mGC-derived exosomes.Exosomes were isolated from pooled serum samples of 20 mGC patients and 40 healthy controls (HCs) by ultracentrifugation. Next, quantitative proteomic analyses were applied to analyze the protein profiles of the exosomes, and bioinformatic analyses were conducted on the proteomic data. Finally, the expression of exosomal protein candidates was selectively validated in individual subjects by western blot analysis. We isolated exosomes from serum samples. The size of the serum derived exosomes ranged from 30 to 150 nm in diameter. The exosomal markers CD9 and CD81 were observed in the serum exosomes. However, the exosomal negative marker calnexin, an endoplasmic reticulum protein, was not detected in exosomes. Overall, 443 exosomal proteins, including 110 differentially expressed proteins (DEPs) were identified by quantitative proteomics analyses. The bioinformatics analyses indicated that the upregulated proteins were enriched in the process of protein metabolic, whereas the downregulated proteins were largely involved in cell-cell adhesion organization. Surprisingly, ten highly vital proteins (UBA52, PSMA1, PSMA5, PSMB6, PSMA7, PSMA4, PSMA3, PSMB1, PSMA6, and FGA) were filtered from DEPs, most of which are proteasome subunits. Moreover, the validation data confirmed that PSMA3 and PSMA6 was explicitly enriched in the serum derived exosomes from patients with mGC. The present study provided a comprehensive description of the serum exosome proteome of mGC patients, which could be an excellent resource for further studies of mGC.

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Serum Exosome Proteome of Tuberculosis Patients Demonstrated Enriched Inflammatory Response Proteins

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