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Proteomic profile of plasma-derived extracellular vesicles from Colombian pregnant women with Plasmodium-soil transmitted helminths coinfection

ABSTRACT: Background: Knowledge about extracellular vesicles (EV) and their molecular cargo in gestational parasitic infections, particularly by Plasmodium and soil-transmitted helminths (STH), is almost non-existent. Objective: To perform isolation and molecular characterization of plasma-derived EVs from Colombian pregnant women and compare quantity, size, concentration and protein cargo of those EVs according to the infectious status. Methodology: Five study groups were formed: 1), Pregnant women with Plasmodium infection. 2), Pregnant women with STH infection. 3), Pregnant women with coinfection Plasmodium and STH. 4), Pregnant women without infection with Plasmodium nor STH. 5), Non-pregnant women without infection with Plasmodium nor STH. Plasma-derived EVs were isolated by size exclusion chromatography (SEC) and fractions containing EVs identified by a bead-based flow cytometric assay for tetraspanin CD9; the size and concentration of EVs were quantified by nanoparticle tracking analysis, and proteins associated with EVs were identified by liquid chromatography-mass spectrometry (LC-MS) in a pool of samples per study group.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Plasma, Extracellular Exosome

SUBMITTER: Eva Borràs

LAB HEAD: Eduard Sabido

PROVIDER: PXD051270 | Pride | 2024-11-26

REPOSITORIES: Pride

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Source:

			Action	DRS
	2023ME000_HEDE_001_004.msf	Msf
	2023ME000_HEDE_001_01_45pto.raw	Raw
	2023ME000_HEDE_002_005.msf	Msf
	2023ME000_HEDE_002_01_45pto.raw	Raw
	2023ME000_HEDE_003_01_45pto.raw	Raw

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Project description:The role of extracellular vesicles (EVs), specifically exosomes, in intercellular communication likely plays a key role in placental orchestration of pregnancy and maternal immune sensing of the fetus. While murine models are powerful tools to study pregnancy and maternal-fetal immune interactions, in contrast to human placental exosomes, the content of murine placental and pregnancy exosomes remains largely understudied. Using a recently developed in vitro culture technique, murine trophoblast stem cells derived from B6 mice were differentiated into syncytial-like cells. EVs from the conditioned media, as well as from pregnant and non-pregnant sera, were enriched for exosomes. The RNA composition of these murine trophoblast-derived and pregnancy-associated exosome-enriched-EVs (ExoE-EVs) was determined using RNA-sequencing analysis and expression levels confirmed by qRT-PCR. Differentially abundant miRNAs were detected in syncytial differentiated ExoE-EVs, particularly from the X chromosome cluster (mmu-miR-322-3p, mmu-miR-322-5p, mmu-miR-503-5p, mmu-miR-542-3p, and mmu-miR-450a-5p). These were confirmed to be increased in pregnant mouse sera ExoE-EVs by qRT-PCR analysis. Interestingly, fifteen miRNAs were only present within the pregnancy-derived ExoE-EVs compared to non-pregnant controls. Mmu-miR-292-3p and mmu-miR-183-5p were noted to be some of the most abundant miRNAs in syncytial ExoE-EVs and were also present at higher levels in pregnant versus non-pregnant sera ExoE-EVs. The bioinformatics tool, MultiMir, was employed to query publicly available databases of predicted miRNA-target interactions. This analysis reveals that the X-chromosome miRNAs are predicted to target ubiquitin-mediated proteolysis and intracellular signaling pathways. Knowing the cargo of placental and pregnancy-specific ExoE-EVs as well as the predicted biological targets informs studies using murine models to examine not only maternal-fetal immune interactions but also the physiologic consequences of placental-maternal communication.

Dataset Information

Proteomic profile of plasma-derived extracellular vesicles from Colombian pregnant women with Plasmodium-soil transmitted helminths coinfection

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