Project description:During embryonic vascular development, endothelial cells that line all blood vessels undergo specification into arterial, capillary, and venous subtypes to form a circulatory network. Regulation of endothelial cell cycle state has been shown to play a critical role in enabling arterial-venous specification and remodeling in a postnatal, blood flow-mediated, and tissue specific manner; however, if a similar cell cycle-mediated mechanism is a common mechanism that regulates embryonic vascular development, even prior to blood flow, is unknown. To investigate this, we first defined the emergence of distinct subtypes by isolating endothelial cells from wild type embryos at embryonic day (E)8.0, prior to blood flow; E8.5, just after flow begins; and E9.5. We performed single cell RNA sequencing and analyses, and found increased specification of arterial, venous and hemogenic subtypes over time, concomitant with decreased primordial and capillary endothelial cells. Gene Ontology analysis revealed that cell cycle control was significantly enriched over time, and we found arterial identity highly correlated with growth arrest. To gain more insight into the potential role of cell cycle control in specification, we isolated similar endothelial cells from Fucci-expressing embryos, sorted them into distinct cell cycle states (early G1, late G1 and S/G2/M), performed bulk RNA sequencing, and bioinformatically correlated endothelial cell cycle states with subtype identities. We found venous endothelial cells are highly enriched in early G1 and arterial endothelial cells highly enriched in late G1, which was corroborated with fluorescent imaging of Fucci embryos. Furthermore, we showed that endothelial cell hyperproliferation, induced by deletion of cell cycle inhibitor Cdkn1b (p27), impaired arterial-venous specification and vascular development. These results support that at the earliest stage of vascular development, endothelial cells in arterial and venous vessels reside in different cell cycle states and endothelial cell cycle control is required for their specification.

Project description:Malaria-naive male rhesus macaques (Macaca mulatta), approximately three years of age, were inoculated intravenously with salivary gland sporozoites produced and isolated at the Centers for Disease Control and Prevention from multiple Anopheles species (An. dirus, An. gambiae, and An. stephensi) and then profiled for clinical, hematological, parasitological, immunological, functional genomic, lipidomic, proteomic, and metabolomic measurements. The experiment was designed for 100 days, and pre- and post-100 day periods to prepare subjects and administer curative treatments respectively. The anti-malarial drug artemether was subcuratively administered selectively to several subjects during the primary parasitemia to suppress clinical complications and to all animals for curative treatment of blood-stage infections to allow detection of relapses. One subject was euthanized during the 100-day experimental period due to clinical complications. The anti-malarial drugs primaquine and chloroquine were administered to all remaining subjects at the end of the study for curative treatment of the liver and blood-stage infections, respectively. Capillary blood samples were collected daily for the measurement of CBCs, reticulocytes, and parasitemias. Capillary blood samples were collected every other day to obtain plasma for metabolomic analysis. Venous blood and bone marrow samples were collected at seven time points for functional genomic, proteomic, lipidomic, and immunological analyses. Within the MaHPIC, this project is known as 'Experiment 04'. This dataset was produced by Lance Wells at UGA. To access other publicly available results from 'E04' and other MaHPIC Experiments, including clinical results (specifics on drugs administered, diet, and veterinary interventions), and other omics, visit http://plasmodb.org/plasmo/mahpic.jsp . This page will be updated as datasets are released to the public. The experimental design and protocols for this study were approved by the Emory University Institutional Animal Care and Use Committee (IACUC). This dataset also includes results from MaHPIC Experiment 18 (E18). LC-MS/MS output from E18 were used as a control for analysis. Refer to the E04 and E18 README files for details of these datasets and their production.

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Strong Concordance of DNA Methylation Measures Between Venous and Capillary Blood - Implications for Human Epigenome Studies

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