Enzymatic Screening and Diagnosis of Lysosomal Storage Diseases

Chunli Yu, MD, Qin Sun, PhD, Hui Zhou, MD, PhD

Abstract


Lysosomal storage diseases (LSDs) are a group of more than 50 genetic disorders.  Clinical symptoms are caused by the deficiency of specific enzyme (enzymes) function and resultant substrate accumulation in the lysosomes, which leads to impaired cellular function and progressive tissue and organ dysfunction.  Measurement of lysosomal enzyme activity plays an important role in the clinical diagnosis of LSDs.  The major enzymatic testing methods include fluorometric assays using artificial 4-methylumbelliferyl (4-MU) substrates, spectrophotometric assays and radioactive assays with radiolabeled natural substrates. As many effective treatment options have become available, presymptomatic diagnosis and early intervention are imperative.  Many methods were developed in the past decade for newborn screening (NBS) of selective LSDs in dried blood spot (DBS) specimens.  Modified fluorometric assays with 4-MU substrates, MS/MS or LC-MS/MS multiplex enzyme assays, digital microfluidic fluorometric assays, and immune-quantification assays for enzyme contents have been reported in NBS of LSDs, each with its own advantages and limitations.  Active technical validation studies and pilot screening studies have been conducted or are ongoing.  These studies have provided insight in the efficacy of various methodologies.   In this review, technical aspects of the enzyme assays used in clinical diagnosis and NBS are summarized.  The important findings from pilot NBS studies are also reviewed.

[N A J Med Sci. 2013;6(4):186-193.   DOI:  10.7156/najms.2013.0604186]



Keywords


lysosomal storage diseases (LSDs), newborn screening (NBS), lysosomal enzyme, tandem mass spectrometry (MS/MS)

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References


Wenger DA, Coppola S, Liu SL. Lysosomal storage disorders: diagnostic dilemmas and prospects for therapy. Genet Med. 2002;4(6):412-419.

Wenger DA, Coppola S, Liu SL. Insights into the diagnosis and treatment of lysosomal storage diseases. Arch Neurol. 2003;60(3):322-328.

Wilcox WR. Lysosomal storage disorders: the need for better pediatric recognition and comprehensive care. J Pediatr. 2004;144(5 Suppl):S3-14.

Wenger DA, Williams C. Screening for Lysosomal Disorders. In: Hommes FA, ed. Techniques on Diagnostic Human Biochemical Genetics. A Laboratory Manual. New York, NY 10003: Wiley-Liss; 1991:587-617.

Meikle PJ, Hopwood JJ, Clague AE, Carey WF. Prevalence of lysosomal storage disorders. JAMA. 1999;281(3):249-254.

van Gelder CM, Vollebregt AA, Plug I, van der Ploeg AT, Reuser AJ. Treatment options for lysosomal storage disorders: developing insights. Expert Opin Pharmacother. 2012;13(16):2281-2299.

Smid BE, Aerts JM, Boot RG, Linthorst GE, Hollak CE. Pharmacological small molecules for the treatment of lysosomal storage disorders. Expert opinion on investigational drugs. 2010;19(11):1367-1379.

Wang RY, Bodamer OA, Watson MS, Wilcox WR. Lysosomal storage diseases: diagnostic confirmation and management of presymptomatic individuals. Genet Med. 2011;13(5):457-484.

Chamoles NA, Blanco M, Gaggioli D. Diagnosis of alpha-L-iduronidase deficiency in dried blood spots on filter paper: the possibility of newborn diagnosis. Clin Chem. 2001;47(4):780-781.

Li Y, Scott CR, Chamoles NA, et al. Direct multiplex assay of lysosomal enzymes in dried blood spots for newborn screening. Clin Chem. 2004;50(10):1785-1796.

Wang D, Eadala B, Sadilek M, et al. Tandem mass spectrometric analysis of dried blood spots for screening of mucopolysaccharidosis I in newborns. Clin Chem. 2005;51(5):898-900.

Duffey TA, Khaliq T, Scott CR, Turecek F, Gelb MH. Design and synthesis of substrates for newborn screening of Maroteaux-Lamy and Morquio A syndromes. Bioorg Med Chem Lett. 2010;20(20):5994-5996.

Duffey TA, Sadilek M, Scott CR, Turecek F, Gelb MH. Tandem mass spectrometry for the direct assay of lysosomal enzymes in dried blood spots: application to screening newborns for mucopolysaccharidosis VI (Maroteaux-Lamy syndrome). Anal Chem. 2010;82(22):9587-9591.

Khaliq T, Sadilek M, Scott CR, Turecek F, Gelb MH. Tandem mass spectrometry for the direct assay of lysosomal enzymes in dried blood spots: application to screening newborns for mucopolysaccharidosis IVA. Clin Chem. 2011;57(1):128-131.

Wolfe BJ, Blanchard S, Sadilek M, Scott CR, Turecek F, Gelb MH. Tandem mass spectrometry for the direct assay of lysosomal enzymes in dried blood spots: application to screening newborns for mucopolysaccharidosis II (Hunter Syndrome). Anal Chem. 2011;83(3):1152-1156.

Wolfe BJ, Ghomashchi F, Kim T, et al. New substrates and enzyme assays for the detection of mucopolysaccharidosis III (Sanfilippo Syndrome) types A, B, C, and D by tandem mass spectrometry. Bioconjug Chem. 2012;23(3):557-564.

Sista RS, Eckhardt AE, Wang T, et al. Digital microfluidic platform for multiplexing enzyme assays: implications for lysosomal storage disease screening in newborns. Clin Chem. 2011;57(10):1444-1451.

Sista RS, Wang T, Wu N, et al. Rapid assays for Gaucher and Hurler diseases in dried blood spots using digital microfluidics. Mol Genet Metab. 2013;109(2):218-220.

Sista RS, Wang T, Wu N, et al. Multiplex newborn screening for Pompe, Fabry, Hunter, Gaucher, and Hurler diseases using a digital microfluidic platform. Clin Chim Acta. 2013;424C:12-18.

Meikle PJ, Grasby DJ, Dean CJ, et al. Newborn screening for lysosomal storage disorders. Mol Genet Metab. 2006;88(4):307-314.

Fuller M, Tucker JN, Lang DL, et al. Screening patients referred to a metabolic clinic for lysosomal storage disorders. J Med Genet. 2011;48(6):422-425.

Voznyi YV, Keulemans JL, van Diggelen OP. A fluorimetric enzyme assay for the diagnosis of MPS II (Hunter disease). J Inherit Metab Dis. 2001;24(6):675-680.

Voznyi Ya V, Karpova EA, Dudukina TV, et al. A fluorimetric enzyme assay for the diagnosis of Sanfilippo disease C (MPS III C). J Inherit Metab Dis. 1993;16(2):465-472.

Wood TC, Harvey K, Beck M, et al. Diagnosing mucopolysaccharidosis IVA. J Inherit Metab Dis. 2013;36(2):293-307.

He W, Voznyi Ya V, Boer AM, Kleijer WJ, van Diggelen OP. A fluorimetric enzyme assay for the diagnosis of Sanfilippo disease type D (MPS IIID). J Inherit Metab Dis. 1993;16(6):935-941.

van Diggelen OP, Voznyi YV, Keulemans JL, et al. A new fluorimetric enzyme assay for the diagnosis of Niemann-Pick A/B, with specificity of natural sphingomyelinase substrate. J Inherit Metab Dis. 2005;28(5):733-741.

Wiederschain G, Raghavan S, Kolodny E. Characterization of 6-hexadecanoylamino-4-methylumbelliferyl-beta-D- galactopyranoside as fluorogenic substrate of galactocerebrosidase for the diagnosis of Krabbe disease. Clin Chim Acta. 1992;205(1-2):87-96.

Baum H, Dodgson KS, Spencer B. The assay of arylsulphatases A and B in human urine. Clin Chim Acta. 1959;4(3):453-455.

Hopwood JJ, Elliott H, Muller VJ, Saccone GT. Diagnosis of Maroteaux-Lamy syndrome by the use of radiolabelled oligosaccharides as substrates for the determination of arylsulphatase B activity. Biochem J. 1986;234(3):507-514.

Kaback MM, Desnick RJ. Hexosaminidase A Deficiency. In: Pagon RA, Adam MP, Bird TD, Dolan CR, Fong CT, Stephens K, eds. GeneReviews. Seattle (WA),1993.

Labrousse P, Chien YH, Pomponio RJ, et al. Genetic heterozygosity and pseudodeficiency in the Pompe disease newborn screening pilot program. Mol Genet Metab. 2010;99(4):379-383.

Zhou H, Fernhoff P, Vogt RF. Newborn bloodspot screening for lysosomal storage disorders. J Pediatr. 2011;159(1):7-13 e11.

The Addition of Pompe Disease to the Recommended Uniform Screening Panel.

http://www.hrsa.gov/advisorycommittees/mchbadvisory/heritabledisorders/recommendations/correspondence/uniformpanel060313.pdf. Accessed on 10/1/2013.

Chamoles NA, Blanco MB, Gaggioli D, Casentini C. Hurler-like phenotype: enzymatic diagnosis in dried blood spots on filter paper. Clin Chem. 2001;47(12):2098-2102.

Chamoles NA, Blanco M, Gaggioli D, Casentini C. Gaucher and

Niemann-Pick diseases--enzymatic diagnosis in dried blood spots on filter paper: retrospective diagnoses in newborn-screening cards. Clin Chim Acta. 2002;317(1-2):191-197.

Chamoles NA, Blanco M, Gaggioli D, Casentini C. Tay-Sachs and Sandhoff diseases: enzymatic diagnosis in dried blood spots on filter paper: retrospective diagnoses in newborn-screening cards. Clin Chim Acta. 2002;318(1-2):133-137.

Chamoles NA, Blanco MB, Iorcansky S, Gaggioli D, Specola N, Casentini C. Retrospective diagnosis of GM1 gangliosidosis by use of a newborn-screening card. Clin Chem. 2001;47(11):2068.

Chamoles NA, Niizawa G, Blanco M, Gaggioli D, Casentini C. Glycogen storage disease type II: enzymatic screening in dried blood spots on filter paper. Clin Chim Acta. 2004;347(1-2):97-102.

Chiang SC, Hwu WL, Lee NC, Hsu LW, Chien YH. Algorithm for Pompe disease newborn screening: results from the Taiwan screening program. Mol Genet Metab. 2012;106(3):281-286.

Chien YH, Hwu WL, Lee NC. Pompe disease: early diagnosis and early treatment make a difference. Pediatrics and neonatology. 2013;54(4):219-227.

Chien YH, Lee NC, Chiang SC, Desnick RJ, Hwu WL. Fabry disease: incidence of the common later-onset alpha-galactosidase A IVS4+919G-->A mutation in Taiwanese newborns--superiority of DNA-based to enzyme-based newborn screening for common mutations. Mol Med. 2012;18:780-784.

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(1):31-40.

Burton B, Charrow J, Angle B, Widera S, Waggoner D. A pilot newborn screening study for lysosomal storage disorders (LSD) in Illinois. Mol Genet Metab. 2012;105:S23-S24.

Blanchard S, Sadilek M, Scott CR, Turecek F, Gelb MH. Tandem mass spectrometry for the direct assay of lysosomal enzymes in dried blood spots: application to screening newborns for mucopolysaccharidosis I. Clin Chem. 2008;54(12):2067-2070.

Li Y, Brockmann K, Turecek F, Scott CR, Gelb MH. Tandem mass spectrometry for the direct assay of enzymes in dried blood spots: application to newborn screening for Krabbe disease. Clin Chem. 2004;50(3):638-640.

Wang D, Wood T, Sadilek M, Scott CR, Turecek F, Gelb MH. Tandem mass spectrometry for the direct assay of enzymes in dried blood spots: application to newborn screening for mucopolysaccharidosis II (Hunter disease). Clin Chem. 2007;53(1):137-140.

Zhang XK, Elbin CS, Chuang WL, et al. Multiplex enzyme assay screening of dried blood spots for lysosomal storage disorders by using tandem mass spectrometry. Clin Chem. 2008;54(10):1725-1728.

la Marca G, Casetta B, Malvagia S, Guerrini R, Zammarchi E. New

strategy for the screening of lysosomal storage disorders: the use of the online trapping-and-cleanup liquid chromatography/mass spectrometry. Anal Chem. 2009;81(15):6113-6121.

Kasper DC, Herman J, De Jesus VR, Mechtler TP, Metz TF, Shushan B. The application of multiplexed, multi-dimensional ultra-high-performance liquid chromatography/tandem mass spectrometry to the high-throughput screening of lysosomal storage disorders in newborn dried bloodspots. Rapid Commun Mass Spectrom. 2010;24(7):986-994.

Duffey TA, Bellamy G, Elliott S, et al. A tandem mass spectrometry triplex assay for the detection of Fabry, Pompe, and mucopolysaccharidosis-I (Hurler). Clin Chem. 2010;56(12):1854-1861.

Orsini JJ, Martin MM, Showers AL, et al. Lysosomal storage disorder 4+1 multiplex assay for newborn screening using tandem mass spectrometry: application to a small-scale population study for five lysosomal storage disorders. Clin Chim Acta. 2012;413(15-16):1270-1273.

Spacil Z, Tatipaka H, Barcenas M, Scott CR, Turecek F, Gelb MH. High-throughput assay of 9 lysosomal enzymes for newborn screening. Clin Chem. 2013;59(3):502-511.

Orsini JJ, Morrissey MA, Slavin LN, et al. Implementation of newborn screening for Krabbe disease: population study and cutoff determination. Clin Biochem. 2009;42(9):877-884.

Duffner PK, Caggana M, Orsini JJ, et al. Newborn screening for Krabbe disease: the New York State model. Pediatr Neurol. 2009;40(4):245-252; discussion 253-245.

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(2):498-503.

Matern D, Oglesbee D, Tortorelli S. Newborn screening for lysosomal storage disorders and other neuronopathic conditions. Dev Disabil Res Rev. 2013;17(3):247-253.

Moss DW. The relative merits and applicability of kinetic and fixed-incubation methods of enzyme assay in clinical enzymology. Clin Chem. 1972;18(12):1449-1454.

De Jesus VR, Zhang XK, Keutzer J, et al. Development and evaluation of quality control dried blood spot materials in newborn screening for lysosomal storage disorders. Clin Chem. 2009;55(1):158-164.

Hwu WL, Chien YH, Lee NC, et al. Newborn screening for Fabry disease in Taiwan reveals a high incidence of the later-onset GLA mutation c.936+919G>A (IVS4+919G>A). Hum Mutat. 2009;30(10):1397-1405.

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(9813):335-341.

Chien YH, Lee NC, Thurberg BL, et al. Pompe disease in infants: improving the prognosis by newborn screening and early treatment. Pediatrics. 2009;124(6):e1116-1125.

Chen LR, Chen CA, Chiu SN, et al. Reversal of cardiac dysfunction after enzyme replacement in patients with infantile-onset Pompe disease. J Pediatr. 2009;155(2):271-275 e272.


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