Relative Distribution of Gb
            <sub>3</sub>
            Isoforms/Analogs in Nod/Scid/Fabry Mice Tissues Determined by Tandem Mass Spectrometry

Provençal, Philippe; Boutin, Michel; Dworski, Shaalee; Au, Bryan; Medin, Jeffrey A.; Auray‐Blais, Christiane

doi:10.4155/bio-2016-0116

Cited by 14 publications

(10 citation statements)

References 42 publications

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“…Mass spectrometry-based proteomics has been recognized as a powerful tool with a potential to uncover detailed changes in protein expression [29]. To date, most of the proteomics studies performed on FD patients examined FD-affected renal tissue or plasma; however, few studies of protein expression have used FD-affected human heart tissue [30,31]. Although it has been revealed that Gb3 accumulation induces endothelial KCa3.1 degradation in Gla -knockout mice through clathrin and Rab5C, which are the critical components of endosome maturation machinery [32], the proteomic profiling performed in our study revealed that GLA knockout led to the downregulation of Rab GTPases, including RAB11 subfamily, which are involved in recycling from an endosomal compartment to the plasma membrane, and was shown to contribute to exosome secretion in neuronal cells [33], although the molecular mechanism of RAB11 function in exosome secretion has yet to be deciphered, especially regarding its downstream effectors.…”

Section: Discussionmentioning

confidence: 99%

Generation of GLA-Knockout Human Embryonic Stem Cell Lines to Model Autophagic Dysfunction and Exosome Secretion in Fabry Disease-Associated Hypertrophic Cardiomyopathy

Song

Chien

Yarmishyn

et al. 2019

Cells

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Fabry disease (FD) is a rare inherited disorder characterized by a wide range of systemic symptoms; it is particularly associated with cardiovascular and renal problems. Enzyme replacement therapy and pharmacological chaperone migalastat are the only approved and effective treatment strategies for FD patients. It is well documented that alpha-galactosidase A (GLA) enzyme activity deficiency causes globotriaosylceramide (Gb3) accumulation, which plays a crucial role in the etiology of FD. However, the detailed mechanisms remain unclear, and the lack of a reliable and powerful disease model is an obstacle. In this study, we created such a model by using CRISPR/Cas9-mediated editing of GLA gene to knockout its expression in human embryonic stem cells (hESCs). The cardiomyocytes differentiated from these hESCs (GLA-null CMs) were characterized by the accumulation of Gb3 and significant increases of cell surface area, the landmarks of FD-associated cardiomyopathy. Furthermore, we used mass spectrometry to compare the proteomes of GLA-null CMs and parental wild type CMs and found that the Rab GTPases involved in exocytotic vesicle release were significantly downregulated. This caused impairment of autophagic flux and protein turnover, resulting in an increase of reactive oxygen species and apoptosis. To summarize, we established a FD model which can be used as a promising tool to study human hypertrophic cardiomyopathy in a physiologically and pathologically relevant manner and to develop new therapies by targeting Rab GTPases signaling-related exosomal vesicles transportation.

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Section: Discussionmentioning

confidence: 99%

Generation of GLA-Knockout Human Embryonic Stem Cell Lines to Model Autophagic Dysfunction and Exosome Secretion in Fabry Disease-Associated Hypertrophic Cardiomyopathy

Song

Chien

Yarmishyn

et al. 2019

Cells

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“…This result indicates that constant digestion by SCDase occurred in both Gb3 isoforms and Gb3 analogs. The presence of Gb3 analogs containing lyso-Gb3 analogs has been suggested in Gla knockout mouse organs (28,29) and the plasma of patients with Fabry disease (19), but this is the first experimental data demonstrating the presence of Gb3 analogs in mouse organs. LC-MS/MS has been widely used to determine Gb3 content in the plasma of patients with Fabry disease (13) and in mouse organs (31,32), and it can detect selected Gb3 isoforms but not Gb3 analogs.…”

Section: Deacylation Of Gb3 From Different Organs Of Gla Tm Tg(cag-a4mentioning

confidence: 92%

“…Next, we assessed whether assaying Gb3 after SCDase deacylation was possible with heterogenous Gb3 containing various isoforms and analogs. As different Gb3 isoforms and analogs have been detected in different mouse organs (28,29), we purified Gb3 from the heart, kidneys, spleen, and liver from Gla tm Tg(CAG-A4GALT) mice. The total Gb3 purified from each organ was weighed, and 4 μg of each preparation was subjected to TLC and visualized with orcinol-sulfuric acid reagent, as previously described (30).…”

Section: Deacylation Of Gb3 From Different Organs Of Gla Tm Tg(cag-a4mentioning

confidence: 99%

Determination of globotriaosylceramide analogs in the organs of a mouse model of Fabry disease

Ishii

Taguchi

Okino

et al. 2020

Journal of Biological Chemistry

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Fabry disease is a heritable lipid disorder caused by the low activity of α-galactosidase A and characterized by the systemic accumulation of globotriaosylceramide (Gb3). Recent studies have reported a structural heterogeneity of Gb3 in Fabry disease, including Gb3 isoforms with different fatty acids and Gb3 analogs with modifications on the sphingosine moiety. However, Gb3 assays are often performed only on the selected Gb3 isoforms. To precisely determine the total Gb3 concentration, here we established two methods for determining both Gb3 isoforms and analogs. One was the deacylation method, involving Gb3 treatment with sphingolipid ceramide N-deacylase, followed by an assay of the deacylated products, globotriaosylsphingosine (lyso-Gb3) and its analogs, by ultra-performance LC coupled to tandem MS (UPLC-MS/MS). The other method was a direct assay established in the present study for 37 Gb3 isoforms and analogs/isoforms by UPLC-MS/MS. Gb3s from the organs of symptomatic animals of a Fabry disease mouse model were mainly Gb3 isoforms and two Gb3 analogs, such as Gb3(+18) containing the lyso-Gb3(+18) moiety and Gb3(−2) containing the lyso-Gb3(−2) moiety. The total concentrations and Gb3 analog distributions determined by the two methods were comparable. Gb3(+18) levels were high in the kidneys (24% of total Gb3) and the liver (13%), and we observed Gb3(−2) in the heart (10%) and the kidneys (5%). These results indicate organ-specific expression of Gb3 analogs, insights that may lead to a deeper understanding of the pathophysiology of Fabry disease.

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“…The MS/MS method can also be used for research purposes to characterize different isoforms of the various SLs in various tissues and samples; indeed, their composition can be variable from one tissue to another (Sevin et al 2007, Provençal et al 2016. Methylated isoforms of Gb 3 are elevated in urine of patients affected with variant FD carrying the p.Asn215Ser mutation (Abaoui et al 2016).…”

Section: Sphingolipidsmentioning

confidence: 99%

Contribution of tandem mass spectrometry to the diagnosis of lysosomal storage disorders

Piraud

Pettazzoni

Lavoie

et al. 2018

J of Inher Metab Disea

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Tandem mass spectrometry (MS/MS) is a highly sensitive and specific technique. Thanks to the development of triple quadrupole analyzers, it is becoming more widely used in laboratories working in the field of inborn errors of metabolism. We review here the state of the art of this technique applied to the diagnosis of lysosomal storage disorders (LSDs) and how MS/MS has changed the diagnostic rationale in recent years. This fine technology brings more sensitive, specific, and reliable methods than the previous biochemical ones for the analysis of urinary glycosaminoglycans, oligosaccharides, and sialic acid. In sphingolipidoses, the quantification of urinary sphingolipids (globotriaosylceramide, sulfatides) is possible. The measurement of new plasmatic biomarkers such as oxysterols, bile acids, and lysosphingolipids allows the screening of many sphingolipidoses and related disorders (Niemann-Pick type C), replacing tedious biochemical techniques. Applied to amniotic fluid, a more reliable prenatal diagnosis or screening of LSDs is now available for fetuses presenting with antenatal manifestations. Applied to enzyme measurements, it allows high throughput assays for the screening of large populations, even newborn screening. The advent of this new method can modify the diagnostic rationale behind LSDs.

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Relative Distribution of Gb ₃ Isoforms/Analogs in Nod/Scid/Fabry Mice Tissues Determined by Tandem Mass Spectrometry

Cited by 14 publications

References 42 publications

Generation of GLA-Knockout Human Embryonic Stem Cell Lines to Model Autophagic Dysfunction and Exosome Secretion in Fabry Disease-Associated Hypertrophic Cardiomyopathy

Generation of GLA-Knockout Human Embryonic Stem Cell Lines to Model Autophagic Dysfunction and Exosome Secretion in Fabry Disease-Associated Hypertrophic Cardiomyopathy

Determination of globotriaosylceramide analogs in the organs of a mouse model of Fabry disease

Contribution of tandem mass spectrometry to the diagnosis of lysosomal storage disorders

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Relative Distribution of Gb 3 Isoforms/Analogs in Nod/Scid/Fabry Mice Tissues Determined by Tandem Mass Spectrometry

Cited by 14 publications

References 42 publications

Generation of GLA-Knockout Human Embryonic Stem Cell Lines to Model Autophagic Dysfunction and Exosome Secretion in Fabry Disease-Associated Hypertrophic Cardiomyopathy

Generation of GLA-Knockout Human Embryonic Stem Cell Lines to Model Autophagic Dysfunction and Exosome Secretion in Fabry Disease-Associated Hypertrophic Cardiomyopathy

Determination of globotriaosylceramide analogs in the organs of a mouse model of Fabry disease

Contribution of tandem mass spectrometry to the diagnosis of lysosomal storage disorders

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Product

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About

Relative Distribution of Gb ₃ Isoforms/Analogs in Nod/Scid/Fabry Mice Tissues Determined by Tandem Mass Spectrometry