Role of extracellular vesicles in trophoblast invasive migration

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The placenta is an essential organ for appropriate foetal development and its successful implantation depends upon invasion of the uterus by trophoblast cells, impacting directly on both animal and human health leading to significant well-being and healthy ageing issues. Epidemiological studies have shown that only around 30% of all conceptions get clinically recognized whilst the majority are lost either spontaneously or during pregnancy complication and that the process of implantation is a key regulator in this process. Despite the importance of the placenta to foetal development, we know little of the mechanisms that mediate placental implantation - the key step in developing a healthy, functional placenta, but the invasive migration of the trophoblast into the receptive endometrium is paramount. This proposed programme of work will provide world-leading, novel molecular mechanistic insight to the process of trophoblast invasive motility during implantation and how it is regulated by microvesicles released from the surrounding endometrium.Extravillous trophoblast cells (EVTs) proliferate and migrate from the cytotrophoblast in the anchoring villi of the placenta and invade the maternal decidua and myometrium. Little is known about the molecular pathways that are important for cellular motility but extracellular vesicle secretion by the endometrium has been shown to be important for such regulation.This project will dissect the molecular mechanisms by which the endometrium regulates trophoblast invasive migration through the release of extracellular vesicles using an array of cellular biology assays that are routinely used in our laboratories to measure cell motility and invasion, and determine the factors involved in these process. The isolation of extracellular vesicles and the factors that are localised within is also routinely done. Human cell lines routinely used will initially be used from endometrial linage to isolate EV and their effects on trophoblast cells for motility and invasion but ex vivo samples will also be tested.In the first part of the project, the best conditions will be established to determine the most responsive trophoblast cells and the most active endometrial extracellular vesicles. The second part of the study will aim to get a full signature of the microvesicles through an unbiased mass spectrometry analysis using protocols and approaches that have been conducted within our current BBSRC-funded work. The last part of the project will aim to characterise some of the newly identified markers and determine whether their presence in extracellular vesicles are important to regulate the trophoblast invasive migration through the use of gain of function and loss of function studies either by gene regulation or through the use of available inhibitors (e.g. antibodies and or chemical inhibitors).