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A molecular roadmap of the aorta-gonad-mesonephros region reveals BMPER as a novel regulator of HSC maturation

ABSTRACT: A molecular roadmap of the aorta-gonad-mesonephros region reveals BMPER as a novel regulator of HSC maturation

PROVIDER: PRJNA399125 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5951920.fastq.gz	Fastqsanger.gz
	SRR5951930.fastq.gz	Fastqsanger.gz
	SRR5951940.fastq.gz	Fastqsanger.gz
	SRR5951950.fastq.gz	Fastqsanger.gz
	SRR5951960.fastq.gz	Fastqsanger.gz

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A molecular roadmap of the aorta-gonad-mesonephros region reveals BMPER as a novel regulator of HSC maturation [OP9]

Project description:In the developing embryo, haematopoietic stem cells (HSCs) emerge from the aorta-gonad-mesonephros (AGM) region but the molecular regulation of this process is poorly understood. Recently, the progression from E9.5 to E10.5 and polarity along the dorso-ventral axis have been identified as clear demarcations of the supportive HSC niche. To identify novel secreted regulators of HSC maturation, we performed RNA-sequencing over these spatio-temporal transitions in the AGM region, and supportive OP9 cell line.

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A molecular roadmap of the aorta-gonad-mesonephros region reveals BMPER as a novel regulator of HSC maturation [AGM]

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Single-cell RNA sequencing of VE-Cadherin+ cells isolated from mouse E10.5 aorta-gonad-mesonephros region.

Project description:The aim of this experiment was to investigate cellular heterogeneity of VE-Cadherin+ cells isolated from mouse E10.5 aorta-gonad-mesonephros region.

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Bulk RNA sequencing of CD44Neg, CD44LoKitNeg, CD44LoKitPos and CD44High populations from mouse Aorta-Gonad-Mesonephros (AGM) region

Project description:The aim of the experiment was to compare newly defined CD44Neg, CD44LoKitNeg, CD44LoKitPos and CD44High populations from mouse Aorta-Gonad-Mesonephros (AGM) region

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RNA sequence of embryonic dorsal skin and AGM (Aorta-gonad-mesonephros) tissues

Project description:RNA sequence of embryonic dorsal skin and AGM (Aorta-gonad-mesonephros) tissues

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Pro-infammatory Aorta-Gonad-Mesonephros-associated macrophages are involved in embryonic development of hematopoietic stem cells

Project description:Hematopoietic stem cells (HSC) are generated from specialized endothelial cells of the embryonic aorta. Previously, inflammatory factors have been implicated in regulating mouse HSC development, but it is unclear what cells in the embryonic aorta-gonad- mesonephros (AGM) microenvironment produce these factors. In the adult, macrophages play both pro- and anti-inflammatory roles. We sought to examine whether macrophages or other hematopoietic cells found in the embryo prior to HSC generation are involved in the AGM HSC-generative microenvironment. Our CyTOF results indicate two abundant myeloid cell types - mannose-receptor positive AGM- associated macrophages (AGM-aM) and mannose-receptor negative macrophages/progenitors. We show that the appearance of macrophages in the AGM is dependent on CX3CR1. AGM-aM express a pro-inflammatory signature, localize to the embryonic aorta and dynamically interact with nascent and emerging intra-aortic hematopoietic cells (IAHC). Importantly, upon macrophage depletion, no adult- repopulating HSCs are detected, thus implicating unique pro-inflammatory AGM- associated macrophages in regulating the embryonic development of HSCs.

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Differentiation of human embryonic stem cells to HOXA+ hemogenic vasculature that resembles the aorta-gonad-mesonephros

Project description:The production of definitive haematopoietic stem/progenitor cells from human pluripotent stem cells (hPSCs) remains a significant challenge. Using reporter lines to track the endothelial (SOX17) to haematopoietic (RUNX1C) transition, we found that hPSC differentiated in growth factor supplemented serum free medium generated RUNX1C+CD34+ clonogenic cells that homed to the bone marrow, but did not engraft. Compared to repopulation-competent cord blood CD34+ cells, RUNX1C+CD34+ progenitors lacked HOXA gene expression, indicating incorrect mesoderm patterning. This deficiency was ameliorated by a timed pulse of WNT activation combined with ACTIVIN antagonism. Significantly, these HOXA+ cultures now formed complex SOX17+ vessels that produced fetal liver-like haematopoietic cells, similar to the human aorta-gonad-mesonephros (AGM). Comparison of transcriptional profiles of these nascent haematopoietic stem/progenitors with cells isolated from human AGM confirmed significant similarities, consistent with the assignment of our in vitro generated cells to the definitive human haematopoietic lineage. Our findings argue that HOXA codes established early in differentiation predict cellular potential and provide correct cell patterning for the specification of definitive haematopoietic lineages from hPSCs.

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Differentiation of human embryonic stem cells to HOXA+ hemogenic vasculature that resembles the aorta-gonad-mesonephros

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Pro-infammatory Aorta-Gonad-Mesonephros-associated macrophages are involved in embryonic development of hematopoietic stem cells

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