Oliver Stone - Development & Cell Biology - 'Understanding cellular heterogeneity in the cardiovascular system'
In this project, you will receive extensive training in cutting edge bioinformatic, single cell genomic, flow cytometry and in vivo gene expression analysis, techniques that are broadly used to address scientific questions across disciplines in academia and industry. Experiments will address two overarching aims that will build on our recent findings. AIM1: Molecular and functional characterization of distinct angioblast populations. We aim to perform a molecular characterisation of distinct angioblast populations and their derivatives using bioinformatic analyses of existing data, single cell multiome analyses of isolated ECs and their progenitors, and histological analyses using in situ hybridisation and immunofluorescence analyses throughout embryonic development. AIM 2: Genetic dissection of angioblast populations using a novel Etv2DreERT2 knock-in mouse model. To determine the functional importance of ECs derived from distinct sources, we will analyse a novel mouse model for intersectional genetics that will allow us to specifically target ECs derived from distinct angioblast populations.
Background
The cardiovascular and lymphatic systems deliver nutrients and remove waste products from each cell in the body, allowing our brains to process information, our hearts to beat and our muscles to move. The innermost layer of blood and lymphatic vessels is formed by a specialised cell type known as endothelial cells (ECs). These cells are critical for normal organ function and tissue repair, and are implicated in a range of pathologies that include heart disease, cancer and Alzheimer’s. In order to meet the demands of the different organs and tissues they serve, ECs acquire unique charactersistics during embryonic development. For example, brain ECs form tight junctions that prevent leakage of fluid, toxins and immune cells into brain tissue, whereas lymphatic ECs form specialised junctions that facilitate immune cell clearance and interstitial fluid absorption from tissues. Despite their importance, we still do not fully understand how ECs acquire these specialised functions. We aim to address this gap in our knowledge, with the overarching goal of engineering specific subtypes of ECs (e.g. lymphatic ECs, liver ECs, brain ECs) from stem cells in vitro. In a developing embryo, ECs are generally thought to differentiate from homogenous pools of progenitors known as angioblasts that acquire heterogeneous traits as they differentiate to form system (e.g. arterial, venous, lymphatic vessels) and organ (e.g. cardiac, liver, brain capillaries) specific networks. In recent years, our work has explored the impact that lineage history has on the terminal fate and function of subtypes of ECs (Stone and Stainier, Dev Cell, 2019, Lupu*, et al., BioRxiv, 2022). We have shown that heterogeneity in endothelial fate and function is dependent on the type of mesoderm from which ECs arise (Stone and Stainier, Dev Cell, 2019); genetic lineage tracing in mice revealed paraxial mesoderm as the overarching progenitor source of lymphatic ECs in most tissues, while blood vessel ECs were shown to arise predominantly from alternative mesodermal sources. Understanding this phenomenon is important, as it will allow us to engineer subtypes of ECs from stem cells in a dish.
Project
In this project, you will receive extensive training in cutting edge bioinformatic, single cell genomic, flow cytometry and in vivo gene expression analysis, techniques that are broadly used to address scientific questions across disciplines in academia and industry. Experiments will address two overarching aims that will build on our recent findings. AIM1: Molecular and functional characterization of distinct angioblast populations. We aim to perform a molecular characterisation of distinct angioblast populations and their derivatives using bioinformatic analyses of existing data, single cell multiome analyses of isolated ECs and their progenitors, and histological analyses using in situ hybridisation and immunofluorescence analyses throughout embryonic development. AIM 2: Genetic dissection of angioblast populations using a novel Etv2DreERT2 knock-in mouse model. To determine the functional importance of ECs derived from distinct sources, we will analyse a novel mouse model for intersectional genetics that will allow us to specifically target ECs derived from distinct angioblast populations.
