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Cardiovascular Research Advance Access first published online on April 29, 2009
This version [Corrected Proof] published online on May 21, 2009
Cardiovascular Research, doi:10.1093/cvr/cvp133
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Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2009. For permissions please email: journals.permissions@oxfordjournals.org.

HoxB5 induces endothelial sprouting in vitro and modifies intussusceptive angiogenesis in vivo involving angiopoietin-2

Stephan Winnik1, Mei Klinkert1, Haymo Kurz2, Christoph Zoeller1, Jennifer Heinke1,4, Yaxu Wu3, Christoph Bode1, Cam Patterson3 and Martin Moser1,*

1 Department of Cardiology, University of Freiburg, Hugstetter Strasse 55, Freiburg 79106, Germany
2 Tissue Dynamics Laboratary, PMU Salzburg, Salzburg, Austria
3 Carolina Cardiovascular Biology Center, University of North Carolina, Chapel Hill, NC, USA
4 Department of Biology, University of Freiburg, Freiburg, Germany

* Corresponding author. Tel: +49 497 6127 03546; fax: +49 497 6127 03254. E-mail adress: martin.moser{at}uniklinik-freiburg.de

Aims: Homeobox (Hox) proteins are transcriptional regulators in embryonic patterning, cell differentiation, proliferation, and migration in vertebrates and invertebrates. A growing body of evidence suggests that Hox proteins are involved in endothelial cell regulation. We have shown earlier that HoxB5 upregulates vascular endothelial growth factor receptor-2 and thereby contributes to enhanced endothelial precursor cell differentiation. Here we aim to elucidate the role of HoxB5 in angiogenesis.

Methods and results: Endothelial cell sprouting was investigated in the human umbilical vein endothelial cell spheroid assay. We investigated in vivo angiogenesis in the chick (Gallus gallus) chorioallantoic membrane assay. Expression profiling of proangiogenic factors was done by quantitative PCR. The angiopoietin-2 (Ang2) promoter and deletion fragments thereof were cloned into the pGL3 reporter system for analysis of transcriptional activity. We observed that HoxB5 enhances endothelial cell sprouting and modulates the expression of adhesion molecules in vitro. Accordingly, we observed a modification of vascular growth by HoxB5 in vivo. The HoxB5 effect is reminiscent of the effects of angiopoietins. We demonstrate that Ang2 is upregulated upon HoxB5 overexpression and that the HoxB5 effect is abolished by the angiopoietin antagonist soluble Tie-2.

Conclusion: HoxB5 has an activating effect on Ang2 that is essential for endothelial cell sprouting and coordinated vascular growth.

KEYWORDS HoxB5; Angiopoietin-2; Angiogenesis

Time for primary review: 32 days


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