Involvement of NDPK-B in glucose metabolism-mediated endothelial damage via activation of the hexosamine biosynthesis pathway and suppression of O-GlcNAcase activity

• Verfasst von: Chatterjee, Anupriya [VerfasserIn]
• Verfasst von: Eshwaran, Rachana [VerfasserIn]
• Verfasst von: Poschet, Gernot [VerfasserIn]
• Verfasst von: Lomada, Santosh K. [VerfasserIn]
• Verfasst von: Halawa, Mahmoud [VerfasserIn]
• Verfasst von: Wilhelm, Kerstin [VerfasserIn]
• Verfasst von: Schmidt, Martina [VerfasserIn]
• Verfasst von: Hammes, Hans-Peter [VerfasserIn]
• Verfasst von: Wieland, Thomas [VerfasserIn]
• Verfasst von: Feng, Yuxi [VerfasserIn]
• Titel: Involvement of NDPK-B in glucose metabolism-mediated endothelial damage via activation of the hexosamine biosynthesis pathway and suppression of O-GlcNAcase activity
• Verf.angabe: Anupriya Chatterjee, Rachana Eshwaran, Gernot Poschet, Santosh Lomada, Mahmoud Halawa, Kerstin Wilhelm, Martina Schmidt, Hans-Peter Hammes, Thomas Wieland and Yuxi Feng
• E-Jahr: 2020
• Jahr: 19 October 2020
• Umfang: 18 S.
• Fussnoten: Gesehen am 26.11.2020
• Titel Quelle: Enthalten in: Cells
• Ort Quelle: Basel : MDPI, 2012
• Jahr Quelle: 2020
• Band/Heft Quelle: 9(2020,10) Artikel-Nummer 2324, 18 Seiten
• ISSN Quelle: 2073-4409
• DOI: doi:10.3390/cells9102324
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: Ang-2
• Sach-SW: nucleoside diphosphate kinase B
• Sach-SW: O-GlcNAcylation
• Sach-SW: OGA
• Sach-SW: UDP-GlcNAc
• K10plus-PPN: 1741165652

Abstract

Our previous studies identified that retinal endothelial damage caused by hyperglycemia or nucleoside diphosphate kinase-B (NDPK-B) deficiency is linked to elevation of angiopoietin-2 (Ang-2) and the activation of the hexosamine biosynthesis pathway (HBP). Herein, we investigated how NDPK-B is involved in the HBP in endothelial cells (ECs). The activities of NDPK-B and O-GlcNAcase (OGA) were measured by in vitro assays. Nucleotide metabolism and O-GlcNAcylated proteins were assessed by UPLC-PDA (Ultra-performance liquid chromatography with Photodiode array detection) and immunoblot, respectively. Re-expression of NDPK-B was achieved with recombinant adenoviruses. Our results show that NDPK-B depletion in ECs elevated UDP-GlcNAc levels and reduced NDPK activity, similar to high glucose (HG) treatment. Moreover, the expression and phosphorylation of glutamine:fructose-6-phosphate amidotransferase (GFAT) were induced, whereas OGA activity was suppressed. Furthermore, overall protein O-GlcNAcylation, along with O-GlcNAcylated Ang-2, was increased in NDPK-B depleted ECs. Pharmacological elevation of protein O-GlcNAcylation using Thiamet G (TMG) or OGA siRNA increased Ang-2 levels. However, the nucleoside triphosphate to diphosphate (NTP/NDP) transphosphorylase and histidine kinase activity of NDPK-B were dispensable for protein O-GlcNAcylation. NDPK-B deficiency hence results in the activation of HBP and the suppression of OGA activity, leading to increased protein O-GlcNAcylation and further upregulation of Ang-2. The data indicate a critical role of NDPK-B in endothelial damage via the modulation of the HBP.