Previous University and Subject: Medical University of Vienna, Human Medicine
Thesis since: 11/2010
The Use of Liquid Chromatography Tandem Mass Spectrometry Methods for High Throughput Newborn Screening and Confirmational Diagnostics of Lysosomal Storage Disorders
In newborn screening (NBS), one major challenges is the implementation of feasible high throughput technologies to daily laboratory routine. The introduction of tandem mass spectrometry (MS/MS) in NBS laboratories has resulted in a rapid expansion of newborn-screening activities in both private and state-sponsored laboratories due to its multi-parametric nature (e.g. in the simultaneous determination of amino acids and acylcarnitines). Consequently, the introduction of MS/MS has unquestionably improved the screening for inborn errors of metabolism, and the technology can be applied to a much wider range of compounds. The interest in expanding the NBS for inborn errors of metabolism for diseases with enzyme function or complex macromolecular targets as biomarkers has increased significantly due to newly developed therapy concepts. First results of clinical trials with new enzyme replacement therapies, substrate reduction therapies, stem cell transplantation and gene therapy indicate that early diagnosis and early therapy introduction have an enormous impact on outcome and efficiency of such therapies. In addition, the information generated by NBS could be used for epidemiological assessments of such diseases as well it might be useful for carrier identification and genetic counseling.
To demonstrate that NBS for such diseases is technically feasible in daily laboratory routine, and to evaluate the incidences of six lysosomal storage disorders in the Austrian population, a 1 year pilot NBS study, will be performed. This study will be run in parallel to the national routine Austrian Newborn Screening Program. Dried blood spot specimens of a one year collective of newborns will be collected consecutively in a one year study period. Samples were analyzed anonymized for enzyme activities of acid β-glucocerebrosidase (M. Gaucher), α-galactosidase (M. Fabry), α-glucosidase (M. Pompe), acid sphingomyelinase (M. Niemann Pick A/B), galactocerebrosidase (M. Krabbe) and α-L-iduronidase (MucopolysaccharidosisTyp 1) by use of specific substrates and multiplex measurement by electrospray tandem mass spectrometry MS/MS. Additional, protocols and approaches will be optimized with new technologies like implementing on-line multi-dimensional chromatography combining sample preparation with sample analysis in order to facilitate sample introduction, ease-of-use and speed of analysis. In line with this, research will be also focused on the expansion of the screening panel with possible additional combinable screening parameters. Potential new screening candidates are: MPS II, MPS IV, MPS VI.
Methods and Skills:
Tendl KA, Schulz SM, Mechtler TP, Bohn A, Metz T, Greber-Platzer S, Kasper DC, Herkner KR, Item CB. DNA methylation pattern of CALCA in preterm neonates with bacterial sepsis as a putative epigenetic biomarker. Epigenetics 8: 1261-1267, 2013
Ostermann KM, Dieplinger R, Lutsch NM, Strupat K, Metz TF, Mechtler TP, Kasper DC. Matrix-assisted laser desorption/ionization for simultaneous quantitation of (acyl-) carnitines and organic acids in dried blood spots. Rapid Commun Mass Spectrom 27: 1497-504, 2013
Kasper DC, Altiok I, Mechtler TP, Böhm J, Straub J, Langgartner M, Pollak A, Herkner KR, Berger A. Molecular detection of late-onset neonatal sepsis in premature infants using small blood volumes: proof-of-concept. Neonatology 103: 268-273, 2013
Mechtler TP, Metz TF, Müller HG, Ostermann K, Ratschmann R, De Jesus VR, Shushan B, Di Bussolo JM, Herman JL, Herkner KR, Kasper DC. Short-incubation mass spectrometry assay for lysosomal storage disorders in newborn and high-risk population screening. J Chromatogr B Analyt Technol Biomed Life Sci 908: 9-17, 2012
Mechtler TP, Stary S, Metz TF, De Jesús VR, Greber-Platzer S, Pollak A, Herkner KR, Streubel B, Kasper DC. Neonatal screening for lysosomal storage disorders: feasibility and incidence from a nationwide study in Austria. Lancet 379: 335-341, 2012
Metz TF, Mechtler TP, Orsini JJ, Martin M, Shushan B, Herman JL, Ratschmann R, Item CB, Streubel B, Herkner KR, Kasper DC. Simplified newborn screening protocol for lysosomal storage disorders. Clin Chem 57: 1286-1294, 2011
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 24: 986-994, 2010
Kasper DC, Mechtler TP, Reischer GH, Witt A, Langgartner M, Pollak A, Herkner KR, Berger A. The bacterial load of Ureaplasma parvum in amniotic fluid is correlated with an increased intrauterine inflammatory response. Diagn Microbiol Infect Dis 67: 117-121, 2010
Kasper DC, Mechtler TP, Böhm J, Petricevic L, Gleiss A, Spergser J, Witt A, Herkner KR, Berger A. In utero exposure to Ureaplasma spp. is associated with increased rate of bronchopulmonary dysplasia and intraventricular hemorrhage in preterm infants. J Perinat Med 39: 331-336, 2011