α-Galactosidase A Deficiency Accelerates Atherosclerosis in Mice With Apolipoprotein E Deficiency

Abstract

Background-␣-Galactosidase A (Gla) deficiency leads to widespread tissue accumulation of neutral glycosphingolipids and is associated with premature vascular complications such as myocardial infarction and stroke. Glycosphingolipids have been shown to accumulate in human atherosclerotic lesions, although their role in atherogenesis is unclear. Methods and Results-To determine whether Gla affects the progression of atherosclerosis, mice were generated with combined deficiencies of apolipoprotein E and Gla. At 45 weeks of age, Gla-deficient mice had developed more atherosclerosis than mice with normal Gla expression (25.1Ϯ14.0 versus 12.3Ϯ9.3 mm 2 of total lesion area, PϽ0.02). This increase in atherosclerosis was associated with the presence of increased Gb3, enhanced inducible nitric oxide synthase expression, and increased nitrotyrosine staining. Conclusions-These findings suggest that deficiency of Gla leads to increased inducible nitric oxide synthase expression and accelerated atherosclerosis. (Fabry disease) is an X-linked disorder that leads to widespread tissue accumulation of neutral glycosphingolipids with ␣-galactosyl linkages consisting primarily of globotriaosylceramide (Gb3). 1 Clinical manifestations of Fabry disease include renal failure, painful neuropathies, angiokeratoma, myocardial infarction, and stroke, which lead to premature mortality. 1 Although premature vascular complications are more common in subjects with Fabry disease, 2-4 the effect of Gla deficiency on atherogenesis is unknown. Glycosphingolipids have been shown to accumulate in atherosclerotic plaques even in subjects without Fabry disease, which suggests they may play a role in atherogenesis. 5 A mouse model of Gla deficiency has been generated by targeted disruption of the Gla gene. 6 These mice accumulate glycosphingolipids in multiple organs with age, including the large blood vessels 7,8 ; thus, they provide a useful model to study the vascular consequences of Gla deficiency. The present study was designed to test the consequences of Gla deficiency in vascular disease with a mouse model of atherosclerosis. Methods MiceTo test the consequences of Gla deficiency on vascular disease, we examined the development of lipid lesions between apolipoprotein E-deficient (ApoE Ϫ/Ϫ ) littermate mice with genetic variation in Gla. ApoE Ϫ/Ϫ mice on the C57BL6/J background were purchased from Jackson Laboratory (Bar Harbor, Me). Gla-deficient (Gla Ϫ/0 or Gla Ϫ/Ϫ ) mice were bred from mice provided by Drs Ashok Kulkarni and Roscoe Brady (National Institutes of Health, Bethesda, Md). The "0" in Gla Ϫ/0 denotes the absence of the second X chromosome of the male mice in this X-linked disease. These mice were backcrossed at least 6 generations to the C57BL6/J strain before being bred to Analysis of AtherosclerosisAt 45 weeks of age, mice were euthanized via exsanguination while under intraperitoneal pentobarbital anesthesia (100 mg/kg). The mice were perfused with phosphate-buffered saline and fixed with formalin with a 25-gauge needle, insert...

Pathological findings in a patient with Fabry disease who died after 2.5 years of enzyme replacement

“…Evidence for excess production of reactive oxygen species, such as superoxide in Fabry disease, has also accumulated [14,15]. Recently, another group has shown that aortic atherosclerosis in mice deficient in both apo E and α-galactosidase A is markedly accelerated compared with either of those mouse models alone [3]. Together, this clinical and experimental evidence suggests that the process of atherosclerosis may be accelerated in Fabry disease.…”

Genomic abnormalities of the murine model of Fabry disease after disease-related perturbation, a systems biology approach

“…The abnormalities in cerebral blood flow, including cerebral hyperperfusion and delayed vascular reactivity to acetazolamide, were reversed by enzyme replacement therapy (ERT) (7)(8)(9). Vascular NO dysregulation, increased nitrotyrosine staining, and accelerated atherosclerosis were recently confirmed in the murine model of Fabry disease (10).…”

An In Vitro Model of Fabry Disease

“…As a result of this work, two models of vascular disease were identified. These models include a heightened arterial thrombotic response after exposure to oxidant injury (7) and enhanced atherosclerosis when bred on an apolipoprotein E1-deficient background (8). These findings are consistent with a macrovascular defect in Fabry disease and suggest that the arterial endothelium should be the focus of studies on the pathophysiology of ␣-Gal A-associated vasculopathy.…”

“…In support of a NO-based mechanism for the macrovascular disease, our group recently reported two abnormalities in the ␣-Gal A null mouse. These abnormalities include a highly robust carotid artery thrombosis in response to oxidant release by rose Bengal (7) and accelerated atherosclerosis in mice bred on an ApoEϪ/Ϫ background (8). In the first model, GlaϪ/0 mice thrombosed their carotid arteries in as little as 15 min after oxidant release.…”