Abstract
Background
Small extracellular vesicles (sEV) play a crucial role in immune responses to viral infection. However, the composition of sEV derived from children with viral pneumonia remains ill defined.
Methods
First, we performed mass spectrometry-based label-free proteomic analysis of urinary sEV in 7 children with viral pneumonia, 4 children with Mycoplasma pneumoniae pneumonia and 20 healthy children. Then a total of 33 proteins were selected to validate by multiple reaction monitoring analysis in an independent cohort of 20 healthy children and 29 children with pneumonia.
Results
In the discovery phase, a total of 1621 proteins were identified, while 260 proteins have differential expression in children with viral pneumonia compared to healthy children. Biological pathways primarily associated with neutrophil degranulation, carbohydrate metabolism and endocytosis were enriched in children with viral pneumonia. Finally, the abundance of eight proteins was verified to be significantly higher in children with viral pneumonia than in healthy children.
Conclusions
This pilot study with proteomic profiles of urinary sEV provided insights to the host response to viral pathogen exposure and potential diagnostic biomarkers for children with viral pneumonia, and served as the basis for understanding the fundamental biology of infection.
Impact
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There were significant differences in the proteomic features of urinary sEV between children with viral pneumonia and those with Mycoplasma pneumoniae pneumonia.
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Many viral infection-related proteins were identified in urinary sEV and overrepresented in children with viral pneumonia, which facilitates our understanding of the fundamental biology of viral infection.
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A total of eight proteins (ANPEP, ASAH1, COL11A1, EHD4, HEXB, LGALS3BP, SERPINA1 and SERPING1) were verified as potential biomarkers for the diagnosis of viral pneumonia in children.
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Data availability
The datasets generated during the current study are available in the ProteomeXchange Consortium repository (www.proteomexchange.org) with the dataset identifier PXD036472.
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Acknowledgements
We are grateful to all the children who took part in the study.
Funding
This study was supported by Grant from the Shanghai Municipal Health Commission (202140111).
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Authors and Affiliations
Contributions
All authors have met the Pediatric Research authorship requirements. L.Y., J.W. and Q.Z. conceived and designed the experiments. J.C., Y.Y., Q.P., J.W. and L.Y. collected clinical samples. J.C., D.J. and S.W. conducted sEV isolation, LC-MS/MS experiments and data analysis. D.J. and Q.Z. drafted the manuscript. All authors reviewed the manuscript.
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Competing interests
D.J., S.W. and Q.Z. are employees of Wayen Biotechnologies, Inc. The other authors declare that they have no conflicts to declare.
Ethics approval and consent to participate
The study procedure was reviewed and approved by the Ethics Committee of Shanghai Children’s Medical Center. Written informed consent was provided by the patients of the children subjects.
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Cheng, J., Ji, D., Yin, Y. et al. Proteomic profiling of urinary small extracellular vesicles in children with pneumonia: a pilot study. Pediatr Res 94, 161–171 (2023). https://doi.org/10.1038/s41390-022-02431-y
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DOI: https://doi.org/10.1038/s41390-022-02431-y
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