Gene expression profiles of control IgG and anti-Dll4 Ab treated retinal endothelial cells (ECs) in EC-specific RiboTag mouse (Cdh5-CreERT2 Rpl22 tm1.1Psam)
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ABSTRACT:
PROVIDER: PRJNA597893 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR10776647.fastq.gz | Fastqsanger.gz | |||
| SRR10776648.fastq.gz | Fastqsanger.gz | |||
| SRR10776649.fastq.gz | Fastqsanger.gz | |||
| SRR10776650.fastq.gz | Fastqsanger.gz | |||
| SRR10776651.fastq.gz | Fastqsanger.gz |
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Project description:We report the gene expression profiles of control IgG and anti-Dll4 Ab treated retinal endothelial cells (ECs) in EC-specific RiboTag mouse (Cdh5-CreERT2 Rpl22 tm1.1Psam )
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Project description:Purpose: Autologous fat grafting is one of the most effective and safest treatment for soft tissue restoration and augmentation, and many efforts has been done to improve its efficiency including adipose tissue derived stem cell (ASC) supplementation. Here, we evaluated the role of Notch ligand Delta-like ligand 4 (Dll4) in angiogenesis in grafted fat and the effect on graft survival using the murine fat graft model, and its impact on the ASC supplementation. Methods: We performed RNA-seq analysis of 3 mouse fat graft groups: 1) control fat grafts, 2) ASC added fat graft, and 3) ASC added fat graft with Dll4 inhibition. The fat grafts were harvested 8 weeks after grafting (n=4), and generated mRNA profiles by deep sequencing using Illumina NovaSeq 6000. Results: We found that when we treated Dll4 blocking antibody to inhibit Dll4, which was highly expressed in endothelial cells (ECs) of grafted fat, angiogenesis in the grafted fat was significantly induced, promoting fat graft retention. In addition, this effect was shown to synergistically improve the fat graft retention when ASCs were concomitantly supplemented. Transcriptome analysis further identified that the expression of the junctional proteins was increased in the ECs and inflammatory processes were downregulated in the grafted fat with ASC supplementation and Dll4 inhibition. Conclusions: This study serves as a basis for developing new potential therapeutic approaches to improve graft retention after cell-assisted transfer by Dll4 inhibition.
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Project description:In hESCs, expression of the Notch ligand DLL4 parallels the emergence of bipotent hematoendothelial progenitors (HEPs) and promotes their hematopoietic differentiation. During differentiation, DLL4 is only expressed in a subpopulation of HEPs. To study the developmental fate of the two subpopulations of HEPs identified by DLL4 expression, we FACS-isolated DLL4high and DLL4low/- HEPs at day 15 of differentiation and performed gene expression analysis using microarrays
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Project description:In hESCs, expression of the Notch ligand DLL4 parallels the emergence of bipotent hematoendothelial progenitors (HEPs) and promotes their hematopoietic differentiation. During differentiation, DLL4 is only expressed in a subpopulation of HEPs. To study the developmental fate of the two subpopulations of HEPs identified by DLL4 expression, we FACS-isolated DLL4high and DLL4low/- HEPs at day 15 of differentiation and performed gene expression analysis using microarrays 10^5 DLL4high and DLL4low/- HEPs purified by FACS from H9 and AND1 hEBs at day 15 of hematopoietic differentiation were used for gene expression analysis using Whole Human Genome Oligo Microarray chips (Agilent Technologies).
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Project description:Endothelial cells (ECs) display organ- and tissue-specific heterogeneity. In the eye, the retinal and choroidal vascular beds are distinct networks with different morphological and functional features with the former representing a tight barrier and the latter, a permeable fenestrated vasculature. Given that retinal health critically relies on the function of these vascular beds and that their dysfunction is implicated in a variety of retinal diseases, a molecular understanding of both physiological and pathophysiological characteristics of these distinct vasculatures is critical. Given their interspersed anatomic distribution among parenchymal cells, the study of EC gene expression, in vivo, has been hampered by the challenge of isolating pure populations of ocular ECs in sufficient quantities for large-scale transcriptomics. To address this challenge, we present a methodological and analytical workflow to facilitate inter-tissue comparisons of the in vivo EC translatome isolated from choroid, retina, and brain using the Cre-inducible NuTRAP flox construct and two widely-used endothelial cre mouse lines, constitutive Tie2-Cre and Tamoxifen-inducible Cdh5-CreERT2, which express EGFP and mCherry tags for cell-specific isolation of nuclei and ribosomes. Within each model, inter-tissue comparison of TRAP-RNAseq enrichment (TRAP translatome vs input transcriptome) showed tissue-specific gene enrichments with differential pathway representation. For each mouse model, inter-tissue comparison of the EC translatome (choroid vs brain, choroid vs retina, and brain vs retina) showed over 50% overlap of differentially expressed genes (DEGs) between the three paired comparisons, with differential pathway representation for each tissue. Pathway analysis of DEGs in the Cdh5-NT vs Tie2-NT comparison for retina, choroid, and brain predicted inhibition of processes related to myeloid cell function and activation, consistent with more specific targeting of ECs in the Cdh5-NT than in the Tie2-NT model. While TRAP enriches for EC transcripts in both models, it also captures myeloid transcripts in the Tie2-NT model. We confirmed these observations with a cell sorting approach. We suggest experimental/analytical considerations should be taken when selecting cre-lines to target ECs.
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