Characterization of a functional endothelial super-enhancer that regulates ADAMTS18 and angiogenesis |
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Authors: | Isidore Mushimiyimana Henri Niskanen Mustafa Beter Johanna
P Laakkonen Minna
U Kaikkonen Seppo Yl-Herttuala Nihay Laham-Karam |
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Institution: | A. I. Virtanen Institute for Molecular Sciences; University of Eastern Finland; Kuopio 70211, Finland;Heart Center and Gene Therapy Unit; Kuopio University Hospital; Kuopio 70029, Finland |
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Abstract: | Super-enhancers are clusters of enhancers associated with cell lineage. They can be powerful gene-regulators and may be useful in cell-type specific viral-vector development. Here, we have screened for endothelial super-enhancers and identified an enhancer from within a cluster that conferred 5–70-fold increase in transgene expression. Importantly, CRISPR/Cas9 deletion of enhancers demonstrated regulation of ADAMTS18, corresponding to evidence of chromatin contacts between these genomic regions. Cell division-related pathways were primarily affected by the enhancer deletions, which correlated with significant reduction in cell proliferation. Furthermore, we observed changes in angiogenesis-related genes consistent with the endothelial specificity of this SE. Indeed, deletion of the enhancers affected tube formation, resulting in reduced or shortened sprouts. The super-enhancer angiogenic role is at least partly due to its regulation of ADAMTS18, as siRNA knockdown of ADAMTS18 resulted in significantly shortened endothelial sprouts. Hence, functional characterization of a novel endothelial super-enhancer has revealed substantial downstream effects from single enhancer deletions and led to the discovery of the cis-target gene ADAMTS18 and its role in endothelial function. |
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