Characterization of follicular fluid-derived extracellular vesicles and their contribution to periconception environment
Date
2022-09-19
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Abstract
Viljatuse levimus on tänapäeval üheks suurimaks murekohaks. Maailma Terviseorganisatsiooni prognooside kohaselt on viljatus 21. sajandil suuruselt kolmas ülemaailmne terviseprobleem, mis jääb alla ainult vähi ja südame-veresoonkonna haigustele. Abistav reproduktiivtehnoloogia (Assisted Reproductive Technology - ART), sealhulgas in vitro viljastamine, on oluline terapeutiline meetod viljatuse ravis. ART edukuse määr ei ole aga endiselt piisavalt kõrgel tasemel ning on veel palju arenguruumi meetodi tõhusamaks muutmisel. Üks peamisi põhjuseid võib olla sugurakkude kvaliteedi tõstmise ebaõnnestumine ning ka ebasobiv implatatsioonieelne keskkond.
Viimastel aastatel on EV-sid (membraaniga ümbritsetud nanosuuruses osakesed) üha enam tunnustatud kui alternatiivset rakkudevahelise suhtluse viisi. Mitmed uuringud on näidanud, et EV-sid on eraldatud peaaegu kõigist bioloogilistest vedelikest, sealhulgas follikulaarvedelikust (FF). FF-st pärinevad EV-d on olulised munarakkude küpsemiseks, viljastamiseks ja embrüo arenguks ning EVd on ka potentsiaalsed patofüsioloogiliste seisundite biomarkerid.
Antud projektis uurisime FF EV-de rolli munajuha geeniekspressiooni reguleerimisel ning nende mõju spermatosoidide elutähtsatele funktsioonidele. Lisaks uurisime ka EV kaasas kantavate komponentide muutuseid polütsüstiliste munasarjade sündroomi (PCOS) patsientide FF EV-des võrreldes tervete inimestega. Meie esimene uuring näitas, et veiste FF ja FF EV-d muudavad veiste munajuhade epiteelirakkude geeniekspressiooni, mis võib kaasa aidata implantatsiooni eelse mikrokeskkonna, viljastamise ja embrüo varajase arengu ettevalmistamisele. Samamoodi näitas meie teine uuring, et veiste FF EV-d parendavad veiste spermatosoidide funktsionaalseid omadusi, eriti elujõulisust, kapatsitatsiooni ja akrosoomireaktsiooni. Lõpuks näitas meie kolmas uuring EV kaasas kantavate komponentide muutuseid erinevates patofüsioloogilistes seisundites (võrreldi PCOS patsiente ja viljakaid naisi) ja nende panust munasarjade signaali häiretesse. MiRNA-de analüüs näitas, et PCOS-i naistelt saadud FF EV-d kannavad tervete naistega võrreldes erinevaid miRNA-sid. Meie tulemused kinnitasid ka PCOS-i naiste puhul uudse FF EV-dest tuletatud miRNA, mida võiks kasutada PCOS-i diagnoosimise potentsiaalse biomarkerina.
Kokkuvõtteks kinnitasid meie uuringute üldised leiud EV-de olemasolu FF-is ja nende mõju spermatosoididele ja munajuhade geeniekspressioonile ning erinevusi FF EV-de kaasas kantavates komponentides PCOS-i patsientide ja tervete naiste puhul.
The prevalence of infertility is one of the most pressing issues in today’s society. Based on forecasts from the World Health Organization, infertility will rank third among the most serious global health problems in the twenty-first century, after cancer and cardiovascular disease. Assisted reproductive technology (ART), which includes in-vitro fertilization (IVF), is an essential therapeutic method for the treatment of infertility. However, the success rate of ART is still not up to the mark and there is abundant room for improvement in ART's. One of the main reasons hindering the success of ART’s could be the failure to enhance the quality of gametes, and undermining the factors present during preconception. Therefore, the current research identifies the bottleneck issues of infertility, methods for gametes improvement, and role of follicular fluid (FF) derived extracellular vesicles (EVs) in enhancing a suitable preconception environment that might be essential for fertilization and early embryo development. In recent years, EVs (membrane-bound nanosized particles) have become increasingly recognized as an alternative mode of intercellular communication. Many published studies have shown that EVs had been isolated from nearly every type of biological fluid, including FF. FF-derived EVs are essential for oocyte maturation, fertilization, embryo development and have potential to act as biomarkers. In this project, we investigated the role of FF EVs in regulating the gene expression of oviduct, its impact on vital functions of spermatozoa, and the cargo changes in FF derived EVs in Polycystic ovarian syndrome (PCOS) patients versus healthy people. Our first study showed that bovine FF and FF EVs alter the gene expression of bovine oviductal epithelial cells, which may contribute to the preparation of preconception microenvironment, fertilization and early embryo development. Similarly, our second study showed that bovine FF EVs enhances the functional properties of bovine spermatozoa, specifically viability, capacitation, and acrosome reaction. Finally, our third study showed EV cargo changes in different pathophysiological condition (PCOS and fertile women) and their contribution to ovarian signal disturbance. The miRNAs analysis revealed that FF EVs derived from PCOS women carry different miRNAs as compared to healthy women. Our results also confirmed novel miRNA derived from FF EVs in case of PCOS women, which could use as a potential biomarker for PCOS diagnosis. In conclusion, our studies overall findings confirmed the presence of EVs in FF and their effects on spermatozoa, and oviductal gene expression, and differences in cargo of FF EVs incase of PCOS patients versus healthy women.
The prevalence of infertility is one of the most pressing issues in today’s society. Based on forecasts from the World Health Organization, infertility will rank third among the most serious global health problems in the twenty-first century, after cancer and cardiovascular disease. Assisted reproductive technology (ART), which includes in-vitro fertilization (IVF), is an essential therapeutic method for the treatment of infertility. However, the success rate of ART is still not up to the mark and there is abundant room for improvement in ART's. One of the main reasons hindering the success of ART’s could be the failure to enhance the quality of gametes, and undermining the factors present during preconception. Therefore, the current research identifies the bottleneck issues of infertility, methods for gametes improvement, and role of follicular fluid (FF) derived extracellular vesicles (EVs) in enhancing a suitable preconception environment that might be essential for fertilization and early embryo development. In recent years, EVs (membrane-bound nanosized particles) have become increasingly recognized as an alternative mode of intercellular communication. Many published studies have shown that EVs had been isolated from nearly every type of biological fluid, including FF. FF-derived EVs are essential for oocyte maturation, fertilization, embryo development and have potential to act as biomarkers. In this project, we investigated the role of FF EVs in regulating the gene expression of oviduct, its impact on vital functions of spermatozoa, and the cargo changes in FF derived EVs in Polycystic ovarian syndrome (PCOS) patients versus healthy people. Our first study showed that bovine FF and FF EVs alter the gene expression of bovine oviductal epithelial cells, which may contribute to the preparation of preconception microenvironment, fertilization and early embryo development. Similarly, our second study showed that bovine FF EVs enhances the functional properties of bovine spermatozoa, specifically viability, capacitation, and acrosome reaction. Finally, our third study showed EV cargo changes in different pathophysiological condition (PCOS and fertile women) and their contribution to ovarian signal disturbance. The miRNAs analysis revealed that FF EVs derived from PCOS women carry different miRNAs as compared to healthy women. Our results also confirmed novel miRNA derived from FF EVs in case of PCOS women, which could use as a potential biomarker for PCOS diagnosis. In conclusion, our studies overall findings confirmed the presence of EVs in FF and their effects on spermatozoa, and oviductal gene expression, and differences in cargo of FF EVs incase of PCOS patients versus healthy women.
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Väitekirja elektrooniline versioon ei sisalda publikatsioone
Keywords
fallopian tubes, epithelial cells, gene expression, spermatozoa, intercellular communication, fertilisation