Article | Published:

Identification of lysosomal and extralysosomal globotriaosylceramide (Gb3) accumulations before the occurrence of typical pathological changes in the endomyocardial biopsies of Fabry disease patients

Abstract

Purpose

Evaluation standards and treatment initiation timing have been debated for a long time, particularly for late-onset Fabry disease (FD), because of its slow progression. However, early initiation of enzyme replacement therapy (ERT) for FD could be effective in stabilizing the disease progression and potentially preventing irreversible organ damage. We aimed to examine globotriaosylceramide (Gb3) deposits in patients’ endomyocardial biopsies to understand the early pathogenesis of FD cardiomyopathy.

Methods

Immunofluorescent (IF) staining of Gb3 and lysosomal-associated membrane protein 1 (LAMP-1) was performed on endomyocardial biopsies of patients suspected of Fabry cardiomyopathy who had negative or only slight Gb3 accumulation determined by toluidine blue staining and electron microscopic examination.

Results

The IF staining results revealed that all patients examined had abundant Gb3 accumulation in their cardiomyocytes, including the ones who are negative for inclusion bodies. Furthermore, we found that early Gb3 deposits were mostly confined within lysosomes, while they appeared extralysosomally at a later stage.

Conclusion

A significant amount of lysosomal Gb3 deposits could be detected by IF staining in cardiac tissue before the formation of inclusion bodies, suggesting the cardiomyocytes might have been experiencing cellular stress and damage early on, before the appearance of typical pathological changes of FD during the disease progression.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    Desnick RJ, Ioannou YA, Eng CM. α-Galactosidase A deficiency: Fabry disease (2001). The Metabolic and Molecular Bases of Inherited Disease. 8th edn. New York: McGraw-Hill; 2001. p. 3373–774.

  2. 2.

    Germain DP. Fabry disease. Orphanet J Rare Dis. 2010;5:30. https://doi.org/10.1186/1750-1172-5-30

  3. 3.

    Bangari DS, Ashe KM, Desnick RJ, et al. alpha-Galactosidase A knockout mice: progressive organ pathology resembles the type 2 later-onset phenotype of Fabry disease. Am J Pathol. 2015;185:651–65. https://doi.org/10.1016/j.ajpath.2014.11.004

  4. 4.

    Desnick RJ, Brady R, Barranger J, et al. Fabry disease, an under-recognized multisystemic disorder: expert recommendations for diagnosis, management, and enzyme replacement therapy. Ann Intern Med. 2003;138:338–46.

  5. 5.

    Nakao S, Kodama C, Takenaka T, et al. Fabry disease: detection of undiagnosed hemodialysis patients and identification of a “renal variant” phenotype. Kidney Int. 2003;64:801–7. https://doi.org/10.1046/j.1523-1755.2003.00160.x

  6. 6.

    Scheidt von W, Eng CM, Fitzmaurice TF, et al. An atypical variant of Fabry’s disease with manifestations confined to the myocardium. N Engl J Med. 1991;324:395–9. https://doi.org/10.1056/NEJM199102073240607

  7. 7.

    Hopkins PV, Campbell C, Klug T, Rogers S, Raburn-Miller J, Kiesling J. Lysosomal storage disorder screening implementation: findings from the first six months of full population pilot testing in Missouri. J Pediatr. 2015;166:172–7. https://doi.org/10.1016/j.jpeds.2014.09.023

  8. 8.

    Inoue T, Hattori K, Ihara K, Ishii A, Nakamura K, Hirose S. Newborn screening for Fabry disease in Japan: prevalence and genotypes of Fabry disease in a pilot study. J Hum Genet. 2013;58:548–52. https://doi.org/10.1038/jhg.2013.48

  9. 9.

    Lin H-Y, Lin HY, Chong K-W, et al. High incidence of the cardiac variant of Fabry disease revealed by newborn screening in the Taiwan Chinese population. Circ Cardiovasc Genet. 2009;2:450–6. https://doi.org/10.1161/CIRCGENETICS.109.862920

  10. 10.

    Mechtler TP, Stary S, Metz TF, et al. Neonatal screening for lysosomal storage disorders: feasibility and incidence from a nationwide study in Austria. Lancet. 2012;379:335–41. https://doi.org/10.1016/S0140-6736(11)61266-X

  11. 11.

    Scott CR, Elliott S, Buroker N, et al. Identification of infants at risk for developing Fabry, Pompe, or mucopolysaccharidosis-I from newborn blood spots by tandem mass spectrometry. J Pediatr. 2013;163:498–503.

  12. 12.

    Spada M, Pagliardini S, Yasuda M, et al. High incidence of later-onset Fabry disease revealed by newborn screening. Am J Hum Genet. 2006;79:31–40. https://doi.org/10.1086/504601

  13. 13.

    Wittmann J, Karg E, Turi S, et al. Newborn screening for lysosomal storage disorders in Hungary. JIMD Rep. 2012;6:117–25. https://doi.org/10.1007/8904_2012_130

  14. 14.

    Desnick RJ, Brady RO. Fabry disease in childhood. J Pediatr. 2004;144(5 Suppl):S20–S26. https://doi.org/10.1016/j.jpeds.2004.01.051

  15. 15.

    Nakao S, Takenaka T, Maeda M, et al. An atypical variant of Fabry’s disease in men with left ventricular hypertrophy. New Engl J Med. 1995;333:288–93. https://doi.org/10.1056/NEJM199508033330504

  16. 16.

    Hsu TR, Hung SC, Chang FP, et al. Later onset Fabry disease, cardiac damage progress in silence: experience with a highly prevalent mutation. J Am Coll Cardiol. 2016;68:2554–63. https://doi.org/10.1016/j.jacc.2016.09.943

  17. 17.

    Askari H, Kaneski CR, Semino-Mora C, et al. Cellular and tissue localization of globotriaosylceramide in Fabry disease. Virchows Arch. 2007;451:823–34. https://doi.org/10.1007/s00428-007-0468-6

  18. 18.

    Khanna R, Soska R, Lun Y, et al. The pharmacological chaperone 1-deoxygalactonojirimycin reduces tissue globotriaosylceramide levels in a mouse model of Fabry disease. Mol Ther. 2010;18:23–33. https://doi.org/10.1038/mt.2009.220

  19. 19.

    Hsu T-R, Sung S-H, Chang F-P, et al. Endomyocardial biopsies in patients with left ventricular hypertrophy and a common Chinese later-onset fabry mutation (IVS4+919G A). Orphanet J Rare Dis. 2014;9:96 https://doi.org/10.1186/1750-1172-9-96

  20. 20.

    Kotani M, Kawashima I, Ozawa H, Ogura K. Generation of one set of murine monoclonal antibodies specific for globo-series glycolipids: evidence for differential distribution of the glycolipids in rat small intestine. Arch Biochem Biophys. 1994;310(1):89–96.

  21. 21.

    Jung SC, Han IP, Limaye A, Xu R. Adeno-associated viral vector-mediated gene transfer results in long-term enzymatic and functional correction in multiple organs of Fabry mice. Proc Natl Acad Sci USA. 2001;98:2676–81.

  22. 22.

    Sakuraba H, Murata-Ohsawa M, Kawashima I, et al. Comparison of the effects of agalsidase alfa and agalsidase beta on cultured human Fabry fibroblasts and Fabry mice. J Hum Genet. 2005;51:180–8. https://doi.org/10.1007/s10038-005-0342-9

  23. 23.

    Putko BN, Wen K, Thompson RB, et al. Anderson-Fabry cardiomyopathy: prevalence, pathophysiology, diagnosis and treatment. Heart Fail Rev. 2015;20:179–91. https://doi.org/10.1007/s10741-014-9452-9

  24. 24.

    Mehta A, Clarke JTR, Giugliani R, et al. Natural course of Fabry disease: changing pattern of causes of death in FOS—Fabry Outcome Survey. J Med Genet. 2009;46:548–52. https://doi.org/10.1136/jmg.2008.065904

  25. 25.

    Biancini GB, Vanzin CS, Rodrigues DB, et al. Globotriaosylceramide is correlated with oxidative stress and inflammation in Fabry patients treated with enzyme replacement therapy. Biochim Biophys Acta. 2012;1822:226–32. https://doi.org/10.1016/j.bbadis.2011.11.001

  26. 26.

    De Francesco PN, Mucci JM, Ceci R, Fossati CA, Rozenfeld PA. Fabry disease peripheral blood immune cells release inflammatory cytokines: role of globotriaosylceramide. Mol Genet Metab. 2013;109:93–99. https://doi.org/10.1016/j.ymgme.2013.02.003

  27. 27.

    Chen K-H, Chien Y, Wang K-L, et al. Evaluation of proinflammatory prognostic biomarkers for Fabry cardiomyopathy with enzyme replacement therapy. Can J Cardiol. 2016;32:1221.e1–1221.e9. https://doi.org/10.1016/j.cjca.2015.10.033

  28. 28.

    Barbey F, Brakch N, Linhart A, et al. Cardiac and vascular hypertrophy in Fabry disease: evidence for a new mechanism independent of blood pressure and glycosphingolipid deposition. Arterioscler Thromb Vasc Biol. 2006;26:839–44. https://doi.org/10.1161/01.ATV.0000209649.60409.38

  29. 29.

    Brakch N, Dormond O, Bekri S, et al. Evidence for a role of sphingosine-1 phosphate in cardiovascular remodelling in Fabry disease. Eur Heart J. 2010;31:67–76.

  30. 30.

    Griffin JL. Infantile acid maltase deficiency. Virchows Arch B. 1984;45:23.

  31. 31.

    Thurberg BL, Lynch Maloney C, Vaccaro C, et al. Characterization of pre- and post-treatment pathology after enzyme replacement therapy for Pompe disease. Lab Invest. 2006;86:1208–20. https://doi.org/10.1038/labinvest.3700484

  32. 32.

    Lim JA, Li L, Raben N. Pompe disease: from pathophysiology to therapy and back again. Front Aging Neurosci. 2014;6:177. https://doi.org/10.3389/fnagi.2014.00177

  33. 33.

    Beer M, Weidemann F, Breunig F, et al. Impact of enzyme replacement therapy on cardiac morphology and function and late enhancement in Fabry’s cardiomyopathy. Am J Cardiol. 2006;97:1515–8. https://doi.org/10.1016/j.amjcard.2005.11.087

  34. 34.

    Kramer J, Niemann M, Störk S, et al. Relation of burden of myocardial fibrosis to malignant ventricular arrhythmias and outcomes in Fabry disease. Am J Cardiol. 2014;114:895–900. https://doi.org/10.1016/j.amjcard.2014.06.019

  35. 35.

    Weidemann F, Niemann M, Breunig F, et al. Long-term effects of enzyme replacement therapy on Fabry cardiomyopathy: evidence for a better outcome with early treatment. Circulation. 2009;119:524–9. https://doi.org/10.1161/CIRCULATIONAHA.108.794529

  36. 36.

    Takenaka T, Teraguchi H, Yoshida A, et al. Terminal stage cardiac findings in patients with cardiac Fabry disease: an electrocardiographic, echocardiographic, and autopsy study. J Cardiol. 2008;51:50–59. https://doi.org/10.1016/j.jjcc.2007.12.001

  37. 37.

    Basso C, Thiene G. Adipositas cordis, fatty infiltration of the right ventricle, and arrhythmogenic right ventricular cardiomyopathy Just a matter of fat?. Cardiovasc Pathol. 2005;14:37–41. https://doi.org/10.1016/j.carpath.2004.12.001

  38. 38.

    Kim E, Choe YH, Han BK, et al. Right ventricular fat infiltration in asymptomatic subjects: observations from ECG-gated 16-slice multidetector CT. J Comput Assist Tomogr. 2007;31:22–28. https://doi.org/10.1097/01.rct.0000236416.05267.6c

  39. 39.

    Fontaine G, Fontaliran F, Zenati O, et al. Fat in the heart. A feature unique to the human species? Observational reflections on an unsolved problem. Acta Cardiol. 1999;54:189–94.

Download references

ACKNOWLEDGMENTS

We would like to thank Dr. Sakuraba for his generosity in sharing the anti-Gb3 monoclonal antibody with us, and Dr. Nien-Jung Chen and Dr. Cheun-Minn Liu for insightful discussions.

This work was supported in part by the Taipei Veterans General Hospital and University System of Taiwan Joint Research Program (VGHUST105-G7-6-1, VGHUST106-G7-3-1 to C.-L. Hsu and D.-M. Niu) and the Ministry of Science and Technology (MOST), Taiwan (MOST-104-2323-B-010-024 to C.-L. Hsu).

Author information

Disclosure

The authors declare no conflicts of interest.

Correspondence to Chia-Lin Hsu PhD or Dau-Ming Niu PhD.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Keywords

  • Fabry disease
  • Cardiomyopathy
  • Globotriaosylceramide
  • Immunofluorescent staining

Further reading

Fig. 1: Magnetic resonance image (MRI) findings of IVS4 FD patients (patients 1, 2, and 4).
Fig. 2: Gb3 immunofluorescent staining of cardiac tissue sections.
Fig. 3: The histological analysis of the IVS4 patients with no or slight Gb3 accumulation in their endomyocardial biopsies.
Fig. 4: Quantification of Gb3 staining fluorescence intensity.
Fig. 5: Colocalization of Gb3 and LAMP-1 immunoactivity in the endomyocardial biopsies of the IVS4 patients 1–5.