Small and Large Extracellular Vesicles From Human Preovulatory Follicular Fluid Display Distinct ncRNA Cargo Profiles and Differential Effects on KGN Granulosa Cells

ABSTRACT Follicular fluid extracellular vesicles (FF EVs) facilitate communication between oocytes and somatic cells within the ovarian follicle, playing a pivotal role in follicular development. This study highlights the molecular and functional distinctions between small (SEV) and large (LEV) FF E...

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Main Authors: Inge Varik, Katariina Johanna Saretok, Kristine Rosenberg, Ileana Quintero, Maija Puhka, Nataliia Volkova, Aleksander Trošin, Paolo Guazzi, Agne Velthut‐Meikas
Format: Article
Language:English
Published: Wiley 2025-07-01
Series:Journal of Extracellular Vesicles
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Online Access:https://doi.org/10.1002/jev2.70119
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author Inge Varik
Katariina Johanna Saretok
Kristine Rosenberg
Ileana Quintero
Maija Puhka
Nataliia Volkova
Aleksander Trošin
Paolo Guazzi
Agne Velthut‐Meikas
author_facet Inge Varik
Katariina Johanna Saretok
Kristine Rosenberg
Ileana Quintero
Maija Puhka
Nataliia Volkova
Aleksander Trošin
Paolo Guazzi
Agne Velthut‐Meikas
author_sort Inge Varik
collection DOAJ
description ABSTRACT Follicular fluid extracellular vesicles (FF EVs) facilitate communication between oocytes and somatic cells within the ovarian follicle, playing a pivotal role in follicular development. This study highlights the molecular and functional distinctions between small (SEV) and large (LEV) FF EV subpopulations, revealing their specialised regulatory roles in granulosa cell (GC) biology and their consequential impact on ovarian function. Single‐EV profiling uncovered distinct tetraspanin distributions, with LEVs containing a lower proportion of CD9/CD63/CD81‐positive particles compared to SEVs. Fluorescent labelling confirmed uptake of both SEVs and LEVs by GCs, supporting their capacity to impact cellular behaviour. Functionally, LEVs increased testosterone production by GCs, whilst SEVs had no effect on steroid hormone secretion, suggesting a specific role for LEVs in androgen biosynthesis. Transcriptomic analysis revealed extensive SEV‐induced changes in GC gene expression, affecting pathways involved in transcription, TGF‐β signalling, extracellular matrix (ECM) remodelling and cell cycle regulation. In contrast, LEVs elicited minimal transcriptional changes, primarily modulating genes associated with immune regulation and oxidative stress defence. Small RNA sequencing further revealed distinct non‐coding RNA (ncRNA) profiles, with SEVs enriched in miRNAs targeting pathways critical for GC differentiation, whilst LEVs carried higher levels of piRNAs implicated in maintaining genomic stability. These findings advance our understanding of FF EV‐mediated intercellular communication and underscore the importance of investigating EV subpopulations independently.
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spelling doaj-art-3eb85d103dae43f1aa96afeb00d915872025-08-20T03:56:49ZengWileyJournal of Extracellular Vesicles2001-30782025-07-01147n/an/a10.1002/jev2.70119Small and Large Extracellular Vesicles From Human Preovulatory Follicular Fluid Display Distinct ncRNA Cargo Profiles and Differential Effects on KGN Granulosa CellsInge Varik0Katariina Johanna Saretok1Kristine Rosenberg2Ileana Quintero3Maija Puhka4Nataliia Volkova5Aleksander Trošin6Paolo Guazzi7Agne Velthut‐Meikas8Department of Chemistry and Biotechnology Tallinn University of Technology Tallinn EstoniaDepartment of Chemistry and Biotechnology Tallinn University of Technology Tallinn EstoniaDepartment of Chemistry and Biotechnology Tallinn University of Technology Tallinn EstoniaHiPREP Core, Institute for Molecular Medicine Finland (FIMM) University of Helsinki Helsinki FinlandHiPREP Core, Institute for Molecular Medicine Finland (FIMM) University of Helsinki Helsinki FinlandDepartment of Chemistry and Biotechnology Tallinn University of Technology Tallinn EstoniaEast Tallinn Central Hospital Tallinn EstoniaHansaBioMed Life Sciences Ltd Tallinn EstoniaDepartment of Chemistry and Biotechnology Tallinn University of Technology Tallinn EstoniaABSTRACT Follicular fluid extracellular vesicles (FF EVs) facilitate communication between oocytes and somatic cells within the ovarian follicle, playing a pivotal role in follicular development. This study highlights the molecular and functional distinctions between small (SEV) and large (LEV) FF EV subpopulations, revealing their specialised regulatory roles in granulosa cell (GC) biology and their consequential impact on ovarian function. Single‐EV profiling uncovered distinct tetraspanin distributions, with LEVs containing a lower proportion of CD9/CD63/CD81‐positive particles compared to SEVs. Fluorescent labelling confirmed uptake of both SEVs and LEVs by GCs, supporting their capacity to impact cellular behaviour. Functionally, LEVs increased testosterone production by GCs, whilst SEVs had no effect on steroid hormone secretion, suggesting a specific role for LEVs in androgen biosynthesis. Transcriptomic analysis revealed extensive SEV‐induced changes in GC gene expression, affecting pathways involved in transcription, TGF‐β signalling, extracellular matrix (ECM) remodelling and cell cycle regulation. In contrast, LEVs elicited minimal transcriptional changes, primarily modulating genes associated with immune regulation and oxidative stress defence. Small RNA sequencing further revealed distinct non‐coding RNA (ncRNA) profiles, with SEVs enriched in miRNAs targeting pathways critical for GC differentiation, whilst LEVs carried higher levels of piRNAs implicated in maintaining genomic stability. These findings advance our understanding of FF EV‐mediated intercellular communication and underscore the importance of investigating EV subpopulations independently.https://doi.org/10.1002/jev2.70119extracellular vesiclesfollicular fluidgranulosa cellsmiRNAncRNAovary
spellingShingle Inge Varik
Katariina Johanna Saretok
Kristine Rosenberg
Ileana Quintero
Maija Puhka
Nataliia Volkova
Aleksander Trošin
Paolo Guazzi
Agne Velthut‐Meikas
Small and Large Extracellular Vesicles From Human Preovulatory Follicular Fluid Display Distinct ncRNA Cargo Profiles and Differential Effects on KGN Granulosa Cells
Journal of Extracellular Vesicles
extracellular vesicles
follicular fluid
granulosa cells
miRNA
ncRNA
ovary
title Small and Large Extracellular Vesicles From Human Preovulatory Follicular Fluid Display Distinct ncRNA Cargo Profiles and Differential Effects on KGN Granulosa Cells
title_full Small and Large Extracellular Vesicles From Human Preovulatory Follicular Fluid Display Distinct ncRNA Cargo Profiles and Differential Effects on KGN Granulosa Cells
title_fullStr Small and Large Extracellular Vesicles From Human Preovulatory Follicular Fluid Display Distinct ncRNA Cargo Profiles and Differential Effects on KGN Granulosa Cells
title_full_unstemmed Small and Large Extracellular Vesicles From Human Preovulatory Follicular Fluid Display Distinct ncRNA Cargo Profiles and Differential Effects on KGN Granulosa Cells
title_short Small and Large Extracellular Vesicles From Human Preovulatory Follicular Fluid Display Distinct ncRNA Cargo Profiles and Differential Effects on KGN Granulosa Cells
title_sort small and large extracellular vesicles from human preovulatory follicular fluid display distinct ncrna cargo profiles and differential effects on kgn granulosa cells
topic extracellular vesicles
follicular fluid
granulosa cells
miRNA
ncRNA
ovary
url https://doi.org/10.1002/jev2.70119
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