Gangqi Wang1, Sarantos Kostidis2, Gesa L Tiemeier1, Wendy M P J Sol1, Margreet R de Vries3, Martin Giera2, Peter Carmeliet4,5, Bernard M van den Berg1, Ton J Rabelink1. 1. From the Division of Nephrology, Department of Internal Medicine (G.W., G.L.T., W.M.P.J.S., B.M.v.d.B., T.J.R.), The Einthoven Laboratory for Vascular and Regenerative Medicine, Leiden University Medical Center, the Netherlands. 2. Center for Proteomics and Metabolomics, Leiden University Medical Center, the Netherlands (S.K., M.G.). 3. Department of Surgery (M.R.d.V.), The Einthoven Laboratory for Vascular and Regenerative Medicine, Leiden University Medical Center, the Netherlands. 4. Department of Oncology, Laboratory of Angiogenesis and Vascular Metabolism, KU Leuven, Vesalius Research Center, VIB, Belgium (P.C.). 5. Laboratory of Angiogenesis and Vascular Metabolism, Vesalius Research Center, VIB, Leuven, Belgium (P.C.).
Abstract
OBJECTIVE: Endothelial cells exposed to laminar shear stress express a thick glycocalyx on their surface that plays an important role in reducing vascular permeability and endothelial anti-inflammatory, antithrombotic, and antiangiogenic properties. Production and maintenance of this glycocalyx layer is dependent on cellular carbohydrate synthesis, but its regulation is still unknown. Approach and Results: Here, we show that biosynthesis of the major structural component of the endothelial glycocalyx, hyaluronan, is regulated by shear. Both in vitro as well as in in vivo, hyaluronan expression on the endothelial surface is increased on laminar shear and reduced when exposed to oscillatory flow, which is regulated by KLF2 (Krüppel-like Factor 2). Using a CRISPR-CAS9 edited small tetracysteine tag to endogenous HAS2 (hyaluronan synthase 2), we demonstrated increased translocation of HAS2 to the endothelial cell membrane during laminar shear. Hyaluronan production by HAS2 was shown to be further driven by availability of the hyaluronan substrates UDP-glucosamine and UDP-glucuronic acid. KLF2 inhibits endothelial glycolysis and allows for glucose intermediates to shuttle into the hexosamine- and glucuronic acid biosynthesis pathways, as measured using nuclear magnetic resonance analysis in combination with 13C-labeled glucose. CONCLUSIONS: These data demonstrate how endothelial glycocalyx function and functional adaptation to shear is coupled to KLF2-mediated regulation of endothelial glycolysis.
OBJECTIVE: Endothelial cells exposed to laminar shear stress express a thick glycocalyx on their surface that plays an important role in reducing vascular permeability and endothelial anti-inflammatory, antithrombotic, and antiangiogenic properties. Production and maintenance of this glycocalyx layer is dependent on cellular carbohydrate synthesis, but its regulation is still unknown. Approach and Results: Here, we show that biosynthesis of the major structural component of the endothelial glycocalyx, hyaluronan, is regulated by shear. Both in vitro as well as in in vivo, hyaluronan expression on the endothelial surface is increased on laminar shear and reduced when exposed to oscillatory flow, which is regulated by KLF2 (Krüppel-like Factor 2). Using a CRISPR-CAS9 edited small tetracysteine tag to endogenous HAS2 (hyaluronan synthase 2), we demonstrated increased translocation of HAS2 to the endothelial cell membrane during laminar shear. Hyaluronan production by HAS2 was shown to be further driven by availability of the hyaluronan substrates UDP-glucosamine and UDP-glucuronic acid. KLF2 inhibits endothelial glycolysis and allows for glucose intermediates to shuttle into the hexosamine- and glucuronic acid biosynthesis pathways, as measured using nuclear magnetic resonance analysis in combination with 13C-labeled glucose. CONCLUSIONS: These data demonstrate how endothelial glycocalyx function and functional adaptation to shear is coupled to KLF2-mediated regulation of endothelial glycolysis.
Authors: Donald A Belcher; Alexander T Williams; Carlos J Munoz; Cynthia R Muller; Cynthia Walser; Andre F Palmer; Pedro Cabrales Journal: J Appl Physiol (1985) Date: 2021-12-16
Authors: Gesa L Tiemeier; Rozemarijn de Koning; Gangqi Wang; Sarantos Kostidis; Rosalie G J Rietjens; Wendy M P J Sol; Sébastien J Dumas; Martin Giera; Cathelijne W van den Berg; Jeroen C J Eikenboom; Bernard M van den Berg; Peter Carmeliet; Ton J Rabelink Journal: Stem Cells Transl Med Date: 2020-03-12 Impact factor: 6.940
Authors: Gangqi Wang; Margreet R de Vries; Wendy M P J Sol; Annemarie M van Oeveren-Rietdijk; Hetty C de Boer; Anton Jan van Zonneveld; Paul H A Quax; Ton J Rabelink; Bernard M van den Berg Journal: Cells Date: 2020-03-29 Impact factor: 6.600