Browsing by Author "Kasvandik, Sergo"

Now showing 1 - 3 of 3

Compartmentalized gene expression profiling of receptive endometrium reveals progesterone regulated ENPP3 is differentially expressed and secreted in glycosylated form
(2016-09) Boggavarapu, Nageswara Rao; Lalitkumar, Sujata; Joshua, Vijay; Kasvandik, Sergo; Salumets, Andres; Lalit Kumar, Parameswaran Grace; Gemzell-Danielsson, Kristina
The complexity of endometrial receptivity at the molecular level needs to be explored in detail to improve the management of infertility. Here, differential expression of transcriptomes in receptive endometrial glands and stroma revealed Ectonucleotide Pyrophosphatase/Phosphodiesterase 3 (ENPP3) as a progesterone regulated factor and confirmed by various methods, both at mRNA and protein level. The involvement of ENPP3 in embryo attachment was tested in an in vitro model for human embryo implantation. Interestingly, there was high expression of ENPP3 mRNA in stroma but not protein. Presence of N-glycosylated ENPP3 in receptive phase uterine fluid in women confirms its regulation by progesterone and makes it possible to use in a non-invasive test of endometrial receptivity.
The role of proteomic changes in endometrial cells – from the perspective of fertility and endometriosis
(2020-05-08) Kasvandik, Sergo; Salumets, Andres, juhendaja; Peters, Maire, juhendaja; Peil, Lauri, juhendaja; Tartu Ülikool. Meditsiiniteaduste valdkond
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.
Stanniocalcin-1 expression in normal human endometrium and dysregulation in endometriosis
(2016) Aghajanova, Lusine; Altmäe, Signe; Kasvandik, Sergo; Salumets, Andres; Stavreus-Evers, Anneli; Giudice, Linda C.
Objective To determine expression of stanniocalcin-1 (STC1) in human endometrium with and without endometriosis and its regulation by steroid hormones. Design Laboratory study. Setting University. Patient(s) Nineteen women with endometriosis and 33 control women. Intervention(s) Endometrial biopsy and fluid sampling. Main Outcome Measure(s) Analysis of early secretory (ESE) and midsecretory (MSE) endometrial secretomes from fertile women with the use of nano–liquid chromatography–dual mass spectrometry; real-time quantitative polymerase chain reaction, and immunohistochemistry for STC1 and its receptor calcium-sensing receptor (CASR) mRNA and proteins in endometrium with and without endometriosis; evaluation of STC1 and CASR mRNA expression in endometrial stromal fibroblasts (eSF) from women with and without endometriosis decidualized with the use of E2P or 8-bromo-cyclic adenosine monophosphate (cAMP). Result(s) STC1 protein was strongly up-regulated in MSE versus ESE in endometrial fluid of fertile women. STC1 mRNA significantly increased in MSE from women with, but not from those without, endometriosis, compared with proliferative endometrium or ESE, with no significant difference throughout the menstrual cycle between groups. STC1 mRNA in eSF from control women increased >230-fold on decidualization with the use of cAMP versus 45-fold from women with endometriosis, which was not seen on decidualization with E2/P. CASR mRNA did not exhibit significant differences in any condition and was not expressed in isolated eSF. STC1 protein immunoexpression in eSF was significantly lower in women with endometriosis compared with control women. Conclusion(s) STC1 protein is significantly up-regulated in MSE endometrial fluid and is dysregulated in eutopic endometrial tissue from women with endometriosis. It is likely regulated by cAMP and may be involved in the pathogenesis of decidualization defects.