Identifying unique regulatory elements related to polymorphic imprinting and gestational aging in the placenta

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The placenta grows during pregnancy and is attached to the wall of the womb. It links the mother to the baby via the umbilical cord. Its main purpose is to supply the baby with all nutrients and oxygen required for growth, whilst removing waste products and carbon dioxide from the baby's blood. If the placenta does not develop or function correctly, this can lead to pregnancy complications that have huge consequences for the baby, not immediately following delivery, but later in life. Cells are the fundamental unit of complex tissues such as the placenta. Cell phenotype and function are very different between the different cell types. It is the code and activity (referred to as expression) of our genes that are ultimately responsible for defining which cells are produced and how they behave. This is achieved by 'epigenetic' modifications to the DNA. One form of epigenetic regulation is achieved by marking certain regions of the genome with methyl groups that tend to act as a semi-permanent block for gene expression. However, DNA methylation does not act alone, there are different interactive mechanisms that help in epigenetic regulation. In recent years a class of genes, known as imprinted genes, have been shown to be essential for placenta development and function. These uniquely regulated genes are epigenetically turned "on" and expressed solely from the maternal or paternal copy, but not both. The placenta is fascinating as it has a unique epigenetic profile having significantly less DNA methylation compared to all other tissues, which subtly changes during pregnancy. The hierarchical consequences of this unique hypomethylated state on the addition layers of epigenetic information have not been investigated. Furthermore, the placenta harbours hundreds of unique imprinted genes not found in other tissues, the expression of which generally decrease with gestation in a DNA methylation independent manner. This suggests other epigenetic mechanisms are responsible for the observed downregulation. Our understanding of this phenomenon has been hindered by the fact we do not know the mechanisms responsible for healthy aging within the placenta. In this application, we seek to understand how methylation in placenta influences other epigenetic marks at different time point in pregnancy and how this ultimately impacts on imprinted gene expression (Objectives 1.1-1.2). To achieve this, we will utilize, and in some cases improve, the latest technologies to identify which area of the genome are associated with different epigenetic signatures and how this influence development (Objectives 2.1 & 3). Furthermore, since we anticipate variability and cell-type specific signatures within our samples, we will profile isolated single-cells, allowing for us to identify patterns which would not be evident in "bulk" placenta biopsies (Objective 2.2). To ensure maximum chance of success building of our previous experiences, we have established a team that includes researchers responsible for technical development at the UK National Capability in Genomics at the Earlham Institute and computational experts for data analysis. Following the analysis using extremely powerful computers and specialised computer programs (Objective 2.3) we will compare our placenta-derived data across time, and within difference cells- types, to identify regions important for gestation-age related expression changes, especially for imprinted genes, which we will subsequently be remove or alter to determine functionality (Objective 4).