témavezető: Rauch Tibor
helyszín (magyar oldal): PTE ÁOK Pécs, Szigeti u. 12. helyszín rövidítés: ÁOK
A kutatási téma leírása:
The feto-maternal immunological compatibility profoundly affects the efficiency of implantation into the endometrium and the outcome of pregnancy. Compromised fetus implantation can have a number of reasons including the miscommunication between blastocyst and implantation ready endometrium. The uterus lining endometrium is characterized by complex morphological changes during menstrual cycle, which process is supervised specific hormones and their nuclear receptors. Steroid nuclear receptors are transcription factors that govern expression of target genes in endometrium to create optimal condition for fetus implantation. Nevertheless, nuclear receptors do not act alone; they interact with epigenetic factors that generate such milieu in cell nucleus wherein the appropriate target genes can be found easily. One of the most frequent epigenetic modifications is DNA methylation, which is associated with gene silencing when it occurs in promoter regions. DNA methylation catalyzing enzymes can interact with nuclear receptors; however, the played roles in implantation are still controversial. The proposed work will investigate how DNA methylation, one of the most frequent epigenetic modifications, can be involved in the very early steps of implantation. In addition, other characteristic epigenetic factors including DNA demethylation and histone-modification involved enzymes will be also investigated. The following three specific aims are proposed:
Specific Aim 1: Exploring Implantation-Associated Expression Pattern of DNA methylation-related genes. Available data and conclusions regarding this question are contradictory. Therefore, we will address the same question by employing new research strategy for clarifying this issue. First, we will establish primary endometrial cell cultures from mouse and explore accompanying gene expression before and after adding blastocysts into the culture. Different variations of this experimental set up will be also investigated. More specifically, we will examine whether adding of hormones (e.g. estrogen, progesterone) and progesterone-induced blocking factor (PIBF) into the primary cell culture can modify expression pattern of DNA methylation–related factors. We will employ quantitative real-time PCR and western blotting for detecting gene expression changes.
Specific Aim 2: Investigation of the Expression of Histone Modifying Factors in Cell Cultures Besides of DNA methylation machinery other epigenetic mechanisms also play pivotal role in the early steps of implantation. Therefore, the expression of a selected set of genes (i.e., emblematic histone acetyltransferases (HATs), histone deacetylases (HDACs), histone methyltransferases (HMTs) and histone demethylases (HDM) will be also investigated using qRT-PCR and western blotting in endometrial cell culture.
Specific aim 3: Characterization of Implantation-Specific Histone Modifying Factors in Mice
The first two specific-aims are devoted to identify implantation-specific epigenetic factors in primary cell cultures. To validate in vitro data, we will investigate the differentially expressed genes in mice. Accordingly, tissue samples will be collected and expression of a selected set of epigenetic factors will be investigated. The proposed methodologies include qRT-PCR, PCR arrays, western blotting and immunofluorescence microscopy.
Overall, this proposal is based on the concept that imbalanced epigenetic regulatory mechanisms might play a decisive role in implantation. This concept also suggests that relatively minor changes in the expression levels of certain genes might have profound effect on implantation, ultimately leading to the compromised implantation.
felvehető hallgatók száma: 1
Jelentkezési határidő: 2021-05-14
2024. IV. 17. ODT ülés Az ODT következő ülésére 2024. június 14-én, pénteken 10.00 órakor kerül sor a Semmelweis Egyetem Szenátusi termében (Bp. Üllői út 26. I. emelet).
Bejelentkezés
