Dataset Information

Proteomics analysis of biological fluids of sleeping sickness patients

ABSTRACT: Background: Human African trypanosomiasis (HAT), also called sleeping sickness, is one of 20 in WHO’s list of ‘Neglected Tropical Diseases’, for which, less than 3,500 cases are reported each year. The disease is caused by the parasite Trypanosoma brucei. Even though the clinical symptoms are well described, physiological aspects of the disease remain unclear. Diagnosis of the disease is difficult, requiring a lumbar puncture to determine the disease stage and guide treatment. Identifying new markers for disease stage has been a challenge for many years. Methods and Findings: In our study, a liquid chromatography coupled with tandem mass spectrometry tandem approach (LC-MS/MS) provides a new way to establish the proteomic profile of patients in stages 1 and 2 of the disease. The biological fluids (serum, cerebrospinal fluid, urine and saliva) of three uninfected controls, three patients with stage 1 disease, and four patients with stage 2 disease, are analyzed. In addition, quantification of samples from 14 controls, 23 patients with stage 1 disease and 43 patients with stage 2 disease by ELISA completes this analysis, and highlights two potential new markers, neuroserpin and moesin, with the latter present in urine — an easily accessed fluid. Conclusion: These result suggest that quantifying proteins present in biological fluids by LC-MS/MS) could provide new biomarkers for the diagnosis and stage discrimination of HAT.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human) Trypanosoma Brucei

TISSUE(S): Urine, Cerebrospinal Fluid, Blood Serum, Saliva

DISEASE(S): Trypanosomiasis

SUBMITTER: Thibaut LEGER

LAB HEAD: Bertrand Courtioux

PROVIDER: PXD007842 | Pride | 2018-11-15

REPOSITORIES: Pride

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Publications

Proteome characterization in various biological fluids of Trypanosoma brucei gambiense-infected subjects.

Bonnet Julien J Garcia Camille C Leger Thibaut T Couquet Marie-Pauline MP Vignoles Philippe P Vatunga Gedeao G Ndung'u Joseph J Boudot Clotilde C Bisser Sylvie S Courtioux Bertrand B

Journal of proteomics 20181107

Human African trypanosomiasis (HAT) is a neglected tropical disease that is endemic in sub-Saharan Africa. Control of the disease has been recently improved by better screening and treatment strategies, and the disease is on the WHO list of possible elimination. However, some physiopathological aspects of the disease transmission and progression remain unclear. We propose a new proteomic approach to identify new targets and thus possible new biomarkers of the disease. We also focused our attenti ...[more]

PMID: 30414516

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Project description:Exosomes are molecular entities derived from membrane vesicles of endocytic origin secreted by most cell types. These vesicles are implicated in cell-to-cell communication, deliver proteins and mRNA molecules between cells. Recent studies have shown that exosomes are found in body fluids such as saliva, blood, urine, amniotic fluid, malignant ascites, bronchoalveolar lavage fluid, synovial fluids and breast milk. Exosomes secreted through human saliva contain mRNA may potentially be useful for diagnostic purposes. Although the exact protective mechanism of saliva RNA is a topic of debate, the consensus is that the enrichment of mRNAs in these nano-vesicles in one of the features of the biomarker discoveries. Our aim was to determine if exosomes are present in human saliva and to nano-characterize their transcriptomic content. Exosomes were purified by differential ultracentrifugation, identified by immunoelectron microscopy, flow cytometry and western blot using a CD-63 antibody. Atomic force microscopy studies revealed ultra structural analysis of both size and density of exosomes. Microarray analysis revealed the presence of 590 mRNA core transcripts are relatively stable inside the exosomes, which can be of saliva mRNA biomarkers. Exosomal mRNA stability was determined by detergent lyses with treatment of RNase. Under in vitro conditions fluorescent dye labeled saliva exosomes were able to communicate between human oral keratinocytes studied by using fluorescence microscopy. The RNA from saliva exosomes can transfer their genetic information to human oral keratinocytes and alters gene expression in the new location. Together, these results suggest that saliva is involved in mRNA trafficking via exosomes, and provides a mechanism for cargoing passenger mRNAs. Our findings are consistent with proposal that exosomes can shuttle RNAs between cells and mRNA is protected inside these vesicles may be a possible resource for biomarker discovery. Experiment Overall Design: Human saliva exosomes were purified through differential centrifugation followed by RNA extraction and hybridization on Affymetrix microarrays. We were able to obtain normal human subjects saliva which are pooled and subjected to ultracentrifugation. The protocol was approved by UCLA Institutional review board. 1 ml of saliva exosomes were used to extract RNA followed by two rounds of amplification by Actorus Amp kit. The amplified RNA was biotin labled and hybridized with Affymetrix protocol.

Dataset Information

Proteomics analysis of biological fluids of sleeping sickness patients

Publications

Proteome characterization in various biological fluids of Trypanosoma brucei gambiense-infected subjects.

Similar Datasets

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