Toward newborn screening of metachromatic leukodystrophy: results from analysis of over 27,000 newborn dried blood spots

Abstract

Purpose

Metachromatic leukodystrophy (MLD) is a lysosomal storage disorder caused by the deficiency of arylsulfatase A (ARSA), which results in the accumulation of sulfatides. Newborn screening for MLD may be considered in the future as innovative treatments are advancing. We carried out a research study to assess the feasibility of screening MLD using dried blood spots (DBS) from de-identified newborns.

Methods

To minimize the false-positive rate, a two-tier screening algorithm was designed. The primary test was to quantify C16:0-sulfatide in DBS by ultraperformance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS). The screening cutoff was established based on the results from 15 MLD newborns to achieve 100% sensitivity. The secondary test was to measure the ARSA activity in DBS from newborns with abnormal C16:0-sulfatide levels. Only newborns that displayed both abnormal C16:0-sulfatide abundance and ARSA activity were considered screen positives.

Results

A total of 27,335 newborns were screened using this two-tier algorithm, and 2 high-risk cases were identified. ARSA gene sequencing identified these two high-risk subjects to be a MLD-affected patient and a heterozygote.

Conclusion

Our study demonstrates that newborn screening for MLD is highly feasible in a real-world scenario with near 100% assay specificity.

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Fig. 1: Sulfatide analysis in DBS.
Fig. 2: Proposed algorithms for the newborn screening of MLD.

References

  1. 1.

    Cesani, M. et al. Mutation update of ARSA and PSAP genes causing metachromatic leukodystrophy. Hum. Mutat. 37, 16–27 (2016).

    CAS  Article  Google Scholar 

  2. 2.

    Gieselmann, V. & Ingeborg, K.-M. Metachromatic Leukodystrophy. In The Online Metabolic and Molecular Bases of Inherited Disease, (eds Valle, D. L., Antonarakis, S. Ballabio, A., Beaudet, A. L. & Mitchell, G. A.) (McGraw-Hill Education: New York, NY, 2019).

  3. 3.

    Martin, H. R. et al. Neurodevelopmental outcomes of umbilical cord blood transplantation in metachromatic leukodystrophy. Biol. Blood Marrow Transplant. 19, 616–624 (2013).

    Article  Google Scholar 

  4. 4.

    Poletti, V. & Biffi, A. Gene-based approaches to inherited neurometabolic diseases. Hum. Gene Ther. 30, 1222–1235 (2019).

    CAS  Article  Google Scholar 

  5. 5.

    Spacil, Z. et al. Sulfatide analysis by mass spectrometry for screening of metachromatic leukodystrophy in dried blood and urine samples. Clin. Chem. 62, 279–286 (2016).

    CAS  Article  Google Scholar 

  6. 6.

    Tan, M. A., Dean, C. J., Hopwood, J. J. & Meikle, P. J. Diagnosis of metachromatic leukodystrophy by immune quantification of arylsulphatase A protein and activity in dried blood spots. Clin. Chem. 54, 1925–1927 (2008).

    CAS  Article  Google Scholar 

  7. 7.

    Hong, X. et al. Leukocyte and dried blood spot arylsulfatase A assay by tandem mass spectrometry. Anal. Chem. 92, 6341–6348 (2020).

    CAS  Article  Google Scholar 

  8. 8.

    Ridsdale, R. et al. Newborn screening (NBS) for metachromatic leukodystrophy (MLD): results from a study of 100,000 deidentified NBS samples. Abstr. Mol. Genet. Metab. 120, S17–S145 (2016).

    Google Scholar 

  9. 9.

    Umapathysivam, K. et al. Determination of acid α-glucosidase protein: evaluation as a screening marker for Pompe disease and other lysosomal storage disorders. Clin. Chem. 46, 1318–1325 (2000).

    CAS  Article  Google Scholar 

  10. 10.

    Harvey, J. S., Carey, W. F. & Morris, C. P. Importance of the glycosylation and polyadenylation variants in metachromatic leukodystrophy pseudodeficiency phenotype. Hum. Mol. Genet. 7, 1215–1219 (1998).

    CAS  Article  Google Scholar 

  11. 11.

    Gieselmann, V., Polten, A., Kreysing, J. & Von Figura, K. Arylsulfatase A pseudodeficiency: loss of a polyadenylylation signal and N-glycosylation site. Proc. Natl. Acad. Sci. A 86, 9436–9440 (1989).

    CAS  Article  Google Scholar 

  12. 12.

    Hopwood, J. J. & Ballabio, A. Multiple Sulfatase Deficiency and the Nature of the Sulfatase Family. In The Online Metabolic and Molecular Bases of Inherited Disease, (eds Valle, D. L., Antonarakis, S., Ballabio, A., Beaudet, A. L. & Mitchell, G. A.) (McGraw-Hill Education, New York, NY, 2019).

  13. 13.

    Liu, Y. et al. Multiplex tandem mass spectrometry enzymatic activity assay for newborn screening of the mucopolysaccharidoses and type 2 neuronal ceroid lipofuscinosis. Clin. Chem. 63, 1118–1126 (2017).

    CAS  Article  Google Scholar 

  14. 14.

    Wenger, D. A. et al. Clinical, pathological, and biochemical studies on an infantile case of sulfatide/GM1 activator protein deficiency. Am. J. Med. Genet. 33, 255–265 (1989).

    CAS  Article  Google Scholar 

  15. 15.

    Gomez-Ospina N. Arylsulfatase A Deficiency. 2006 May 30 [Updated 2020 Apr 30]. In GeneReviews® (eds Adam M. P. et al.) [Internet]. (University of Washington, Seattle, 1993-2020).

  16. 16.

    Hettiarachchi, D. & Dissanayake, V. H. W. Three novel variants in the arylsulfatase A (ARSA) gene in patients with metachromatic leukodystrophy (MLD). BMC Res. Notes 12, 726 (2019).

    CAS  Article  Google Scholar 

  17. 17.

    Eng, B. et al. Identification of nine novel arylsulfatase a (ARSA) gene mutations in patients with metachromatic leukodystrophy (MLD). Hum. Mutat. 22, 418–419 (2003).

    Article  Google Scholar 

  18. 18.

    Shukla, P. et al. Molecular and structural analysis of metachromatic leukodystrophy patients in Indian population. J. Neurol. Sci. 301, 38–45 (2011).

    CAS  Article  Google Scholar 

  19. 19.

    Wolf, N. I. et al. Metachromatic leukodystrophy and transplantation: remyelination, no cross-correction. Ann. Clin. Transl. Neurol. 7, 169–180 (2020).

    CAS  Article  Google Scholar 

  20. 20.

    Narayanan, D. L. et al. Spectrum of ARSA variations in Asian Indian patients with arylsulfatase A deficient metachromatic leukodystrophy. J. Hum. Genet. 64, 323–331 (2019).

    CAS  Article  Google Scholar 

  21. 21.

    Barth, M. L., Fensom, A. & Harris, A. Identification of seven novel mutations associated with metachromatic leukodystrophy. Hum. Mutat. 6, 170–176 (1995).

    CAS  Article  Google Scholar 

  22. 22.

    Chace, D. H. & Hannon, W. H. Impact of second-tier testing on the effectiveness of newborn screening. Clin. Chem. 56, 1653–1655 (2010).

    CAS  Article  Google Scholar 

  23. 23.

    Langan, T. J. et al. Development of a newborn screening tool based on bivariate normal limits: using psychosine and galactocerebrosidase determination on dried blood spots to predict Krabbe disease. Genet. Med. 21, 1644–1651 (2018).

    Article  Google Scholar 

  24. 24.

    Tortorelli, S. et al. Moonlighting newborn screening markers: the incidental discovery of a second-tier test for Pompe disease. Genet. Med. 20, 840–846 (2018).

    CAS  Article  Google Scholar 

  25. 25.

    Turgeon, C. T. et al. Determination of total homocysteine, methylmalonic acid, and 2-methylcitric acid in dried blood spots by tandem mass spectrometry. Clin. Chem. 56, 1686–1695 (2010).

    CAS  Article  Google Scholar 

  26. 26.

    Chang, M. H. et al. Saposins A, B, C, and D in plasma of patients with lysosomal storage disorders. Clin. Chem. 46, 167–174 (2000).

    CAS  Article  Google Scholar 

  27. 27.

    Krivit, W. Allogeneic stem cell transplantation for the treatment of lysosomal and peroxisomal metabolic diseases. Springer Semin. Immunopathol. 26, 119–132 (2004).

    Article  Google Scholar 

  28. 28.

    Ott, R., Waye, J. S., Chang, P. L. & Chang, P. Evolutionary origins of two tightly linked mutations in arylsulfatase-A pseudodeficiency. Hum. Genet. 101, 135–140 (1997).

    CAS  Article  Google Scholar 

  29. 29.

    Hohenschutz, C. et al. Probable metachromatic leukodystrophy/pseudodeficiency compound heterozygote at the arylsulfatase A locus with neurological and psychiatric symptomatology. Am. J. Med. Genet. 31, 169–175 (1988).

    CAS  Article  Google Scholar 

  30. 30.

    Hopkins, P. V. et al. Incidence of 4 lysosomal storage disorders from 4 years of newborn screening. JAMA Pediatr. 172, 696–697 (2018).

    Article  Google Scholar 

  31. 31.

    Burton, B. K. et al. Newborn screening for lysosomal storage disorders in Illinois: the initial 15-month experience. J. Pediatr. 190, 130–135 (2017).

    Article  Google Scholar 

  32. 32.

    Lee, S. et al. Evaluation of X-linked adrenoleukodystrophy newborn screening in North Carolina. JAMA Netw. Open 3, e1920356 (2020).

    Article  Google Scholar 

  33. 33.

    Scott, C. R. et al. Newborn screening for mucopolysaccharidoses: results of a pilot study with 100 000 dried blood spots. J. Pediatr. 216, 204–207 (2019).

    Article  Google Scholar 

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Acknowledgements

We are grateful for Giancarlo la Marca for providing part of the MLD newborn DBS used in the study. Funding is provided by Takeda and National Institutes of Health (R01 DK067859).

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Correspondence to Michael H. Gelb PhD.

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M.H.G. is a consultant for PerkinElmer Corp. PerkinElmer was not involved in any aspects of the study described in this paper. The other authors declare no conflicts of interest.

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Hong, X., Daiker, J., Sadilek, M. et al. Toward newborn screening of metachromatic leukodystrophy: results from analysis of over 27,000 newborn dried blood spots. Genet Med (2020). https://doi.org/10.1038/s41436-020-01017-5

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