Fabry disease: renal sphingolipid distribution in the α-Gal A knockout mouse model by mass spectrometric and immunohistochemical imaging

Abstract

Fabry disease is an X-linked lysosomal storage disease due to deficient α-galactosidase A (α-Gal A) activity and the resultant lysosomal accumulation of globotriaosylceramide (Gb3) and related lipids primarily in blood vessels, kidney, heart, and other organs. The renal distribution of stored glycolipid species in the α-Gal A knockout mouse model was compared to that in mice to assess relative distribution and absolute amounts of accumulated sphingolipid isoforms. Twenty isoforms of five sphingolipid groups were visualized by mass spectrometry imaging (MSI), and their distribution was compared with immunohistochemical (IHC) staining of Gb3, the major stored glycosphingolipid in consecutive tissue sections. Quantitative bulk lipid analysis of tissue sections was assessed by electrospray ionization with tandem mass spectrometry (ESI-MS/MS). In contrast to the findings in wild-type mice, all three analytical techniques (MSI, IHC, and ESI-MS/MS) revealed increases in Gb3 isoforms and ceramide dihexosides (composed mostly of galabiosylceramides), respectively. To our knowledge, this is the first report of the distribution of individual molecular species of Gb3 and galabiosylceramides in kidney sections in Fabry disease mouse. In addition, the spatial distribution of ceramides, ceramide monohexosides, and sphingomyelin forms in renal tissue is presented and discussed in the context of their biosynthesis.

Glycosphingolipid Storage in Fabry Mice Extends Beyond Globotriaosylceramide and is Affected by ABCB1 Depletion

“…A study of only Gb 3 and DHC acyl chain species was recently reported, but the analyses in that study were limited to the kidney and utilized a different Gla -deficient mouse [54]. Another study has shown sex differences of Gb 3 species levels in Fabry and WT mouse kidney and urine [55].…”

“…The greatest number of significant changes was seen in the kidney, with five of the six species being elevated. A recent study has also shown increases in kidney DHCs in the Fabry mouse [54]. The authors of that study state that galabiosylceramides comprise the majority of DHCs in the kidney.…”

“…The accumulation of MHCs is particularly intriguing given that levels of most Fabry mouse DHCs were unchanged (and some even decreased) in comparison to WT. A previous study looking at renal GSLs showed MHC species to generally be unchanged in the Fabry mouse kidney; however, as mentioned, this study used a different Fabry mouse model and performed analyses at 70 weeks age [54]. …”

Mass Spectrometry Imaging: A Review of Emerging Advancements and Future Insights

“…216 Lipids also have the potential to show tumor boundaries, specifically in breast cancer. 217 Because of their diverse structure, several common ionization sources, including MALDI, 1,20,42,97 DESI, 22,217,218 SIMS, 219–221 and IR-MALDESI, 18,19 have found success in lipid imaging. Notably, newly developed laser spray ionization (LSI), with its simple sample preparation, is becoming more commonly used for lipids as well.…”

MALDI mass spectrometric imaging meets “omics”: recent advances in the fruitful marriage

“…Peptides FT-ICR (positive) 79 Automatic nozzle sprayer 70 Human colorectal cancer tissue IM (positive-V-mode) 80 Binary matrix of 2,5dihydroxybenzoic acid and cyano-4-hydoxycinnamic acid …”