Characterization and microRNA quantification of plasma-derived extracellular vesicles in patients with Plasmodium...

Area of Science:

• Biochemistry
• Molecular Biology
• Parasitology

Background:

• Extracellular vesicles (EVs) mediate intercellular communication and contain microRNAs (miRNAs) that can serve as diagnostic markers.
• Plasmodium knowlesi malaria poses a significant health challenge, necessitating improved diagnostic tools.

Purpose of the Study:

• To investigate the potential of plasma-derived EV-miRNAs as diagnostic biomarkers for Plasmodium knowlesi infection.
• To explore the role of specific miRNAs in the pathogenesis of P. knowlesi.

Main Methods:

• Isolation and characterization of EVs from Plasmodium knowlesi-infected patients and healthy controls using size exclusion chromatography and ultracentrifugation.
• Quantification of EV concentration and size using nanoparticle tracking analysis and transmission electron microscopy.
• Measurement of human miRNA expression levels (hsa-miR-223-5p, hsa-miR-486-5p) via reverse transcriptase quantitative polymerase chain reaction.

Main Results:

• EVs from infected patients were larger and more concentrated than those from controls.
• Hsa-miR-223-5p and hsa-miR-486-5p were significantly upregulated in P. knowlesi-infected patients.
• Bioinformatic analysis indicated that these miRNAs target human and parasite genes involved in malaria-related pathways.
• High areas under the receiver operating characteristic curve (0.9154 for hsa-miR-223-5p, 0.8231 for hsa-miR-486-5p) suggest diagnostic potential.

Conclusions:

• Plasma-derived EV-miRNAs, specifically hsa-miR-223-5p and hsa-miR-486-5p, show potential as non-invasive diagnostic biomarkers for P. knowlesi infection.
• These miRNAs may play a role in regulating gene expression during P. knowlesi infection.
• Further research is warranted to explore their therapeutic potential.