Abstract
DNA ja RNA-põhiste meetodite kasutamine on reproduktiivmeditsiini arengut märkimisväärselt mõjutanud, suurendades arusaamist haiguste pärilikest tagamaadest ja tuues kliinilisse praktikasse uusi diagnostilisi meetodeid. Geenide peamiseks funktsionaalseks väljundiks on aga valgud ning nende uurimine on ainus viis haiguslikest protsessidest vahetu ja detailsema pildi saamiseks. Seda enam, et esmase geeniekspressiooni seos valkude tasemetega rakkudes, kudedes ja kehavedelikes on tihtilugu piiratud. Valkude uurimiseks kasutatav massispektromeetrial põhinev proteoomika on kiirelt arenev teadusharu, mis võimaldab bioloogilisest materjalist, st. nii kudedest, rakkudest kui kehavedelikest kõrge tundlikkusega määrata nende valgulist koostist ehk proteoomi. Antud uurimistöö eesmärgiks oligi rakendada kaasaegseid massispektromeetria-põhiseid proteoomika meetodeid, et leida täpsemaid vastuseid naiste reproduktiivtervisega seotud probleemide molekulaarsete mehhanismide selgitamiseks ja välja pakkuda uusi markerikandidaate diagnostikaks. Täpsemalt keskendus meie töö endometrioosile ja kehavälise viljastamise (IVF) efektiivsusega seotud biomarkerite otsingule.
Meie tööst selgus, et endometrioosi korral on haiguskolde rakkudes toimunud mitmeid muutusi, mis sarnanevad vähi- ja tüvirakkudes kirjeldatud ainevahetuslike kohastumustega. Meie tulemused näitavad, et endometrioosikolde rakud kasutavad sõltumata hapniku juuresolust suurenenud määral anaeroobset ainevahetust. Samuti on kolderakkudes muutunud valkude avaldumine, mis on seotud rakkude levimise ja ellujäämisega. Meie tulemused pakuvad küll välja potentsiaalseid sihtmärk-valke haiguskulu mõjutamiseks, kuid rõhutavad ka vajadust edasisteks uuringuteks suuremates kohortides. Töö teises osas uurisime emakaõõne sekreedi valke, et leida biomarkereid kehavälise viljastamise optimeerimiseks. Selle töö tulemusena leidsime, et emakaõõne sekreet sisaldab valke, mida on võimalik rakendada embrüosiirdamise efektiivsemaks ajastamiseks. Samuti viitavad meie tulemused, et naistel, kellel on IVF korduvalt ebaõnnestunud, esineb häireid emaka limaskesta vastuvõtlikkuses embrüotele ehk retseptiivsuses. Leitud valguliste biomarkerite paneeli on võimalik arendada diagnostiliseks testiks, mis tõstaks IVFi tõhusust ja aitaks paremini tuvastada retseptiivsushäirega naisi.
The impact of genomics and transcriptomics methods for reproductive medicine has been substantial by increasing the understanding behind heritable factors of diseases and by enabling new avenues for diagnostic testing. Nevertheless, the main functional output of genes are proteins, and, studying proteins is essential for providing further detail into the understanding of pathogenetic mechanisms. Furthermore, the correlation between early gene expression and the levels of proteins in cells, tissues and bodily fluids has been shown to be limited. Mass spectrometry-based proteomics is a progressively developing field that has finally reached to a point where measurement of proteomes or the full protein complement is now becoming feasible. The main objective of the current work was to apply modern proteomics methods to enhance the understanding behind reproductive diseases and to propose new biomarkers for diagnostics. More specifically, our work focused on endometriosis and the search for markers that could improve the in vitro fertilization (IVF) process. By analyzing cells from the uterine lining and endometriosis lesions, we found evidence for altered energy metabolism in endometriotic stromal cells and these changes are akin to changes previously seen in tumor and stem cells. In addition, lesion stromal cells appear to have changes in the expression of proteins that are associated with increased invasiveness and survival of cells. Our results propose novel candidates for future studies to see whether targeting these proteins could affect the course of the illness. In the second part of our work, we analyzed secreted proteins from uterine cells to find biomarkers for improving the success rates of IVF. As a result of our study, we found that the uterine secretome contains proteins that reflect the embryo receptivity status of the endometrium. Our results also underlined that women with recurrent implantation failure in IVF have altered uterine secretomes. The proposed biomarker panel of our study can be developed into a diagnostic test that would increase the effectiveness of the IVF procedure and would help to screen for women with pathological uterine receptivity.
The impact of genomics and transcriptomics methods for reproductive medicine has been substantial by increasing the understanding behind heritable factors of diseases and by enabling new avenues for diagnostic testing. Nevertheless, the main functional output of genes are proteins, and, studying proteins is essential for providing further detail into the understanding of pathogenetic mechanisms. Furthermore, the correlation between early gene expression and the levels of proteins in cells, tissues and bodily fluids has been shown to be limited. Mass spectrometry-based proteomics is a progressively developing field that has finally reached to a point where measurement of proteomes or the full protein complement is now becoming feasible. The main objective of the current work was to apply modern proteomics methods to enhance the understanding behind reproductive diseases and to propose new biomarkers for diagnostics. More specifically, our work focused on endometriosis and the search for markers that could improve the in vitro fertilization (IVF) process. By analyzing cells from the uterine lining and endometriosis lesions, we found evidence for altered energy metabolism in endometriotic stromal cells and these changes are akin to changes previously seen in tumor and stem cells. In addition, lesion stromal cells appear to have changes in the expression of proteins that are associated with increased invasiveness and survival of cells. Our results propose novel candidates for future studies to see whether targeting these proteins could affect the course of the illness. In the second part of our work, we analyzed secreted proteins from uterine cells to find biomarkers for improving the success rates of IVF. As a result of our study, we found that the uterine secretome contains proteins that reflect the embryo receptivity status of the endometrium. Our results also underlined that women with recurrent implantation failure in IVF have altered uterine secretomes. The proposed biomarker panel of our study can be developed into a diagnostic test that would increase the effectiveness of the IVF procedure and would help to screen for women with pathological uterine receptivity.
Description
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Keywords
reproductive medicine, endometrium, endometriosis, proteomics, biomarkers