Establishing 3-nitrotyrosine as a biomarker for the vasculopathy of Fabry disease.

Abstract

The endothelial dysfunction of Fabry disease results from α-galactosidase A deficiency leading to the accumulation of globotriaosylceramide. Vasculopathy in the α-galactosidase A null mouse is manifested as oxidant-induced thrombosis, accelerated atherogenesis, and impaired arterial reactivity. To better understand the pathogenesis of Fabry disease in humans, we generated a human cell model by using RNA interference. Hybrid endothelial cells were transiently transfected with small interfering RNA (siRNA) specifically directed against α-galactosidase A. Knockdown of α-galactosidase A was confirmed using immunoblotting and globotriaosylceramide accumulation. Endothelial nitric oxide synthase (eNOS) activity was correspondingly decreased by >60%. Levels of 3-nitrotyrosine (3NT), a specific marker for reactive nitrogen species and quantified using mass spectrometry, increased by 40- to 120-fold without corresponding changes in other oxidized amino acids, consistent with eNOS-derived reactive nitrogen species as the source of the reactive oxygen species. eNOS uncoupling was confirmed by the observed increase in free plasma and protein-bound aortic 3NT levels in the α-galactosidase A knockout mice. Finally, 3NT levels, assayed in biobanked plasma samples from patients with classical Fabry disease, were over sixfold elevated compared with age- and gender-matched controls. Thus, 3NT may serve as a biomarker for the vascular involvement in Fabry disease.

• Authors: Liming Shu, Anuradha Vivekanandan-Giri, Subramaniam Pennathur, Bouwien E Smid, Johannes M F G Aerts, Carla E M Hollak, James A Shayman
• Journal: Kidney international (Kidney Int) Vol. 86 Issue 1 Pg. 58-66 (Jul 2014) ISSN: 1523-1755 [Electronic] United States
• PMID: 24402087 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Biomarkers
• RNA, Small Interfering
• 3-nitrotyrosine
• Tyrosine
• NOS3 protein, human
• Nitric Oxide Synthase Type III
• GLA protein, human
• alpha-Galactosidase

Topics

• Adolescent
• Adult
• Animals
• Biomarkers (metabolism)
• Case-Control Studies
• Cell Line
• Disease Models, Animal
• Endothelial Cells (metabolism)
• Fabry Disease (complications, genetics, metabolism)
• Human Umbilical Vein Endothelial Cells
• Humans
• Male
• Mice
• Mice, Inbred C57BL
• Mice, Knockout
• Middle Aged
• Nitric Oxide Synthase Type III (metabolism)
• RNA, Small Interfering (genetics)
• Tyrosine (analogs & derivatives, metabolism)
• Vascular Diseases (etiology, metabolism)
• Young Adult
• alpha-Galactosidase (antagonists & inhibitors, genetics)