Abstract
BackgroundAdaptation to low oxygen by changing gene expression is vitally important for cell survival and tissue development. The sprouting of new blood vessels, initiated from endothelial cells, restores the oxygen supply of ischemic tissues. In contrast to the transcriptional response induced by hypoxia, which is mainly mediated by members of the HIF family, there are only few studies investigating alternative splicing events. Therefore, we performed an exon array for the genome-wide analysis of hypoxia-related changes of alternative splicing in endothelial cells.Methodology/Principal findingsHuman umbilical vein endothelial cells (HUVECs) were incubated under hypoxic conditions (1% O2) for 48 h. Genome-wide transcript and exon expression levels were assessed using the Affymetrix GeneChip Human Exon 1.0 ST Array. We found altered expression of 294 genes after hypoxia treatment. Upregulated genes are highly enriched in glucose metabolism and angiogenesis related processes, whereas downregulated genes are mainly connected to cell cycle and DNA repair. Thus, gene expression patterns recapitulate known adaptations to low oxygen supply. Alternative splicing events, until now not related to hypoxia, are shown for nine genes: six which are implicated in angiogenesis-mediated cytoskeleton remodeling (cask, itsn1, larp6, sptan1, tpm1 and robo1); one, which is involved in the synthesis of membrane-anchors (pign) and two universal regulators of gene expression (cugbp1 and max).Conclusions/SignificanceFor the first time, this study investigates changes in splicing in the physiological response to hypoxia on a genome-wide scale. Nine alternative splicing events, until now not related to hypoxia, are reported, considerably expanding the information on splicing changes due to low oxygen supply. Therefore, this study provides further knowledge on hypoxia induced gene expression changes and presents new starting points to study the hypoxia adaptation of endothelial cells.
Highlights
The sufficient supply of all cells of the body with oxygen is of vital importance and vessel growth is essential to promote blood supply after tissue ischemia
Confluent Human umbilical vein endothelial cells (HUVECs) were incubated in triplicates for 48 h under normoxic (21% O2) or hypoxic (1% O2) conditions
Total RNA was isolated and tested for vegfa mRNA expression, to confirm the response of the endothelial cells to hypoxia. qRT-PCR analysis showed a 6-fold induction of vegfa mRNA levels under hypoxic compared to normoxic conditions (Figure 1A)
Summary
The sufficient supply of all cells of the body with oxygen is of vital importance and vessel growth is essential to promote blood supply after tissue ischemia. The proliferation, migration and sprouting of endothelial cells from existing capillaries, a process termed angiogenesis, increases blood supply in ischemic tissues [1,2]. Hypoxia leads to the stabilization of the HIF1A protein and thereby initiates an orchestrated signaling response affecting the expression of direct and indirect target genes. Adaptation to low oxygen by changing gene expression is vitally important for cell survival and tissue development. The sprouting of new blood vessels, initiated from endothelial cells, restores the oxygen supply of ischemic tissues. In contrast to the transcriptional response induced by hypoxia, which is mainly mediated by members of the HIF family, there are only few studies investigating alternative splicing events. We performed an exon array for the genome-wide analysis of hypoxia-related changes of alternative splicing in endothelial cells
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
