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A Regulatory microRNA Network Controls Endothelial Cell Phenotypic Switch During Sprouting Angiogenesis [shDICER]

ABSTRACT: A Regulatory microRNA Network Controls Endothelial Cell Phenotypic Switch During Sprouting Angiogenesis [shDICER]

PROVIDER: PRJNA476874 | ENA |

REPOSITORIES: ENA

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A Regulatory microRNA Network Controls Endothelial Cell Phenotypic Switch During Sprouting Angiogenesis [2D]

Project description:We exploited a three-dimensional model of sprouting angiogenesis in which the phenotypic switch from quiescent ECs to tip cells was triggered by VEGF-A. The information obtained about modulated miRNAs and their related protein-coding gene targets was used to generate a co-expression network encompassing gene modules that are post-transcriptionally regulated and whose activity is required for sprouting.

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A Regulatory microRNA Network Controls Endothelial Cell Phenotypic Switch During Sprouting Angiogenesis [shDICER]

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A Regulatory microRNA Network Controls Endothelial Cell Phenotypic Switch During Sprouting Angiogenesis

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A Regulatory microRNA Network Controls Endothelial Cell Phenotypic Switch During Sprouting Angiogenesis [miRNA-seq]

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A regulatory microRNA network controls endothelial cell phenotypic switch during sprouting angiogenesis.

Project description:Angiogenesis requires the temporal coordination of the proliferation and the migration of endothelial cells. Here, we investigated the regulatory role of microRNAs (miRNAs) in harmonizing angiogenesis processes in a three-dimensional in vitro model. We described a microRNA network which contributes to the observed down- and upregulation of proliferative and migratory genes, respectively. Global analysis of miRNA-target gene interactions identified two sub-network modules, the first organized in upregulated miRNAs connected with downregulated target genes and the second with opposite features. miR-424-5p and miR-29a-3p were selected for the network validation. Gain- and loss-of-function approaches targeting these microRNAs impaired angiogenesis, suggesting that these modules are instrumental to the temporal coordination of endothelial migration and proliferation. Interestingly, miR-29a-3p and its targets belong to a selective biomarker that is able to identify colorectal cancer patients who are responding to anti-angiogenic treatments. Our results provide a view of higher-order interactions in angiogenesis that has potential to provide diagnostic and therapeutic insights.

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Cx40 Suppresses Sprouting Angiogenesis In Vitro

Project description:Blood vessels are highly organized and form during development through a series of complex processes that include vasculogenesis, sprouting angiogenesis, and vessel remodeling. Several gap junction proteins (termed connexins, Cx) – including Cx40 (GJA5) – are expressed in vascular endothelium early during vessel development and are critical for establishment of a healthy vasculature. However, Cx40’s specific role in regulating vessel growth remains uncertain: while previous studies have shown that developmental and cancer-associated neovascularization is reduced in Cx40-knockout mice, Cx40 knockout in zebrafish embryos enhances intersegmental vessel growth. Thus, in the current study, our aim was to identify Cx40’s specific role in sprouting angiogenesis. First, we used a vessel-on-a-chip microphysiological model to confirm Cx40’s overall necessity for microvessel network development. Next, we used the fibrin gel bead assay – a three-dimensional in vitro model of sprouting angiogenesis – to assess Cx40’s necessity for this process. We found that Cx40 knockdown in endothelial cells drives more aggressive sprouting angiogenesis in association with increased endothelial cell proliferation. By contrast, using Electrical Cell-substrate Impedance Sensing (ECIS) we observed no effect of Cx40 knockdown on endothelial cell migration. Lastly, we found that Cx37 (GJA4) is reduced in Cx40- deficient endothelial cells, and that targeted silencing of Cx37 alone produces a more aggressive, hyper-sprouting phenotype compared to control or Cx40 knockdown endothelial cells. Taken together, our data argue that Cx40 plays multiple roles during vessel growth, including to specifically limit sprouting angiogenesis, and that this may occur (at least in part) through regulation of endothelial Cx37 levels.

2023-12-14 | GSE243404 | GEO

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