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Transcriptomics analysis investigate the molecular mechanisms of carnosol on t-BHP induced Human retinal microvascular endothelial cells (HRMECs)

ABSTRACT: Transcriptomics analysis investigate the molecular mechanisms of carnosol on t-BHP induced Human retinal microvascular endothelial cells (HRMECs)

PROVIDER: PRJNA773964 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR16550176_1.fastq.gz	Fastqsanger.gz
	SRR16550176_2.fastq.gz	Fastqsanger.gz
	SRR16550177_1.fastq.gz	Fastqsanger.gz
	SRR16550177_2.fastq.gz	Fastqsanger.gz
	SRR16550178_1.fastq.gz	Fastqsanger.gz

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Transcriptomics analysis investigate the molecular mechanisms of carnosol on t-BHP induced Human retinal microvascular endothelial cells (HRMECs)

Project description:Purpose: Transcriptomics analysis provides a genome-wide digital gene expression profile, which reveals the regulation of gene transcription at the overall level. This study aimed to investigate the underlying mechanism of carnosol against t-BHP-induced Human retinal microvascular endothelial cells (HRMECs) oxidative stress injury. Methods: HRMECs were treated with 10 μM carnosol for 24 h, followed by 300 μM t-BHP for 4 h to simulate the oxidative stress environment in vitro. Oligo dT was used to generate the cDNA library. After constructing the cDNA library, the quality of the library was detected with Agilent biological analyzer, and the cDNA was sequenced with Illumina. Conclusions: Our data provide a systematic perspective for the mechanism of carnosol against HRMECs oxidative stress injury, which lay a foundation for further study on carnosol.

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CUT & Tag analysis of primary human retinal microvascular endothelial cells

Project description:CUT & Tag analysis was conducted using primary human retinal microvascular endothelial cells treated with 0 or 100ng/ml VEGF.

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Novel insights into retinal microvascular endothelial cells revealed through single cell RNA-seq analysis.

Project description:Purpose: To better characterise the population structure of primary bovine retinal microvascular endothelial cells (RMECs) based upon their individual transcriptomes. Methods: Individual RMECs were captured on the Fluidigm C1 system (Fluidigm), cDNA libraries were prepared using a Nextera XT kit and sequencing performed on a NextSeq (Illumina). Results: Application of a single cell RNA-seq analysis workflow showed that RMECs form a relatively homogeneous population in culture, with the main subgroup being proliferating cells. Expression of markers from along the arteriovenous tree suggests that most cells originate from capillaries. An in silico model of the blood retinal barrier was created, including junctional proteins not previously reported within the retinal vasculature. Numerous alternative splicing events involving exons within microvascular barrier genes were observed and in many cases individual cells expressed exclusively one isoform. Conclusions: We have optimised a workflow for single-cell transcriptomics in primary RMECs. Our results have provided fundamental insights into the genes involved in retinal microvascular barrier formation.

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CUT & Tag analysis of primary human retinal microvascular endothelial cells

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Effect of ADAM10 depletion in human retinal microvascular endothelial cells (HRMVECs)

Project description:Effect of ADAM10 depletion in human retinal microvascular endothelial cells (HRMVECs)

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Prostaglandin F2α promotes human retinal microvascular endothelial cell proliferation and migration

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Reduced Retinal Microvascular Density, Improvement in Forepaw Reach, Comparative Microarray and Gene Set Enrichment Analysis using DNA Enzyme Targeting c-jun mRNA

Project description:Retinal neovascularization is a critical component in the pathogenesis of common ocular disorders that cause blindness, and treatment options are limited. We evaluated the therapeutic effect of a DNA enzyme targeting c-jun mRNA in mice with pre-existing retinal neovascularization. A single injection of Dz13 in a lipid formulation containing N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl-sulfate and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine inhibited c-Jun expression and reduced retinal microvascular density. The DNAzyme inhibited retinal microvascular density as effectively as VEGF-A antibodies. Comparative microarray and gene expression analysis determined that Dz13 suppressed not only c-jun but a range of growth factors and matrix-degrading enzymes. Dz13 in this formulation inhibited microvascular endothelial cell proliferation, migration and tubule formation in vitro. Moreover, animals treated with Dz13 sensed the top of the cage in a modified forepaw reach model, unlike mice given a DNAzyme with scrambled RNA-binding arms that did not affect c-Jun expression. These findings demonstrate reduction of microvascular density and improvement in forepaw reach in mice administered catalytic DNA. Total RNA from pooled eyes was used to prepare labeled probes for microarrays. Pooled samples included a Control, Vehicle, Dz13 and Dz13_scrambled.

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