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Transcriptomics of retinal endothelial cells isolated from mice at Postnatal day 6.

ABSTRACT: Transcriptomics of retinal endothelial cells isolated from mice at Postnatal day 6.

PROVIDER: PRJNA821837 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR18558257.fastq.gz	Fastqsanger.gz
	SRR18558258.fastq.gz	Fastqsanger.gz
	SRR18558259.fastq.gz	Fastqsanger.gz
	SRR18558260.fastq.gz	Fastqsanger.gz

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Transcriptomics of retinal endothelial cells isolated from mice at Postnatal day 6.

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Project description:Angiogenesis, a process mediating the expansion of vascular beds in many physiological and pathological settings, requires dynamic changes in endothelial cell (EC) behavior. The molecular mechanisms governing EC activity during different phases of vascular growth, remodeling, maturation, and quiescence remain elusive. Here, we have employed actively translating transcriptome analysis of mouse retinal ECs for the characterization of dynamic gene expression changes during postnatal development and the identification of critical angiogenic factors. In silico computational analyses of these data enabled the identification of candidate regulators controlling EC behavior at different developmental checkpoints. The detailed characterization of Mafb, one of the identified candidates, established that this transcription factor controls endothelial sprouting in vitro and in vivo. Integrative analysis of RNA-Seq and ChIP-Seq data defined putative direct Mafb targets, which are repressed or activated by the transcriptional regulator. Together, our results identify novel cell-autonomous regulatory mechanisms controlling sprouting angiogenesis.

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Project description:Super-selective intra-ophthalmic artery chemotherapy (SSIOAC) is an organ-specific drug-delivery strategy to treat retinoblastoma, the most common primary ocular malignancy in children. Unfortunately, recent clinical reports associate adverse vascular toxicities with SSIOAC using melphalan, the most commonly used chemotherapeutic. To explore the reason for the unexpected vascular toxicities, we have developed in vitro studies with human retinal endothelial cells to test the effects of the chemotherapeutics and a non-human primate model to monitor the SSIOAC treatment in real-time and post-treatment. Melphalan and carboplatin (another chemotherapeutic used to treat retinoblastoma via SSIOAC) triggered migration, proliferation, and apoptosis when used to treat human retinal endothelial cells. Melphalan was associated with increased adhesion of leukocytes to human retinal endothelial cells, and tended to increase with increased cell expression of adhesion proteins (ICAM-1) and soluble chemotactic factors (IL-8). Histopathology post-SSIOAC indicated vessel wall sloughing, leukostasis, and vessel occlusion. We have established an in vitro human cell culture model and a non-human primate model to evaluate strategies designed to obviate vascular side effects, and optimize the efficacy of SSIAOC and the drug preparations used in SSIOAC. 4 non-treated (CNT) vs. 4 carboplatin-treated primary human retinal endothelial cells (RECs).

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