Skip to main content
SpringerLink
Log in

Prevalence of tetrahydrobiopterine (BH4)-responsive alleles among Austrian patients with PAH deficiency: comprehensive results from molecular analysis in 147 patients

  • Original Article
  • Published:
Journal of Inherited Metabolic Disease

Abstract

Phenylketonuria (PKU, MIM 261600) is an autosomal recessive disorder caused by mutations of the phenylalanine hydroxylase gene (PAH, GenBank U49897.1, RefSeq NM_000277). To date more than 560 variants of the PAH gene have been identified. In Europe there is regional distribution of specific mutations. Due to recent progress in chaperone therapy, the prevalence of BH4-responsive alleles gained therapeutic importance. Here we report the mutational spectrum of PAH deficiency in 147 unrelated Austrian families. Overall mutation detection rate was 98.6 %. There was a total of 62 disease-causing mutations, including five novel mutations IVS4 + 6T>A, p.H290Y, IVS8-2A>G, p.A322V and p.I421S. The five most prevalent mutations found in patients were p.R408W, IVS12 + 1G>A, p.R261Q, p.R158Q and IVS2 + 5G>C. Neonatal phenylalanine levels before treatment were available in 114/147 patients. Prediction of BH4-responsiveness in patients with full genotypes was exclusively made according to published data. Among the 133 patients needing dietary treatment, 28.4 % are expected to be BH4 "non-responsive", 4.5 % are highly likely BH4-responsive, 35.8 % are probably BH4-responsive while no interpretation was possible for 31.3 %. The mutation data reflect the population history of Austria and provide information on the likely proportion of Austrian PKU patients that may benefit from BH4-therapy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Anjema K, Venema G, Hofstede FC, Carbasius Weber EC, Bosch AM, Ter Horst NM, Hollak CE, Jonkers CF, Rubio-Gozalbo ME, van der Ploeg EM, de Vries MC, Janssen-Regelink RG, Janssen MC, Zweers-van Essen H, Boelen CC, van der Herberg-van de Wetering NA, Heiner-Fokkema MR, van Rijn M, van Spronsen FJ (2011) The 48-hour tetrahydrobiopterin loading test in patients with phenylketonuria: evaluation of protocol and influence of baseline phenylalanine concentration. Mol Genet Metab 104 Suppl:60–63

    Article  Google Scholar 

  • Bernegger C, Blau N (2002) High frequency of tetrahydrobiopterin-responsiveness among hyperphenylalaninemias: a study of 1,919 patients observed from 1988 to 2002. Mol Genet Metab 77(4):304–313

    Article  PubMed  CAS  Google Scholar 

  • Blau N, Erlandsen H (2004) The metabolic and molecular bases of tetrahydrobiopterin-responsive phenylalanine hydroxylase deficiency. Mol Genet Metab 82(2):101–111

    Article  PubMed  CAS  Google Scholar 

  • Blau N, Belanger-Quintana A, Demirkol M, Feillet F, Giovannini M, MacDonald A, Trefz FK, van Spronsen FJ (2009) Optimizing the use of sapropterin (BH(4)) in the management of phenylketonuria. Mol Genet Metab 96(4):158–163

    Article  PubMed  CAS  Google Scholar 

  • Dahri S, Desviat LR, Perez B, Leal F, Ugarte M, Chabraoui L (2010) Mutation analysis of phenylketonuria patients from Morocco: high prevalence of mutation G352fsdelG and detection of a novel mutation p.K85X. Clin Biochem 43(1–2):76–81

    Article  PubMed  CAS  Google Scholar 

  • Daniele A, Cardillo G, Pennino C, Carbone MT, Scognamiglio D, Correra A, Pignero A, Castaldo G, Salvatore F (2007) Molecular epidemiology of phenylalanine hydroxylase deficiency in Southern Italy: a 96 % detection rate with ten novel mutations. Ann Hum Genet 71(2):185–193

    Article  PubMed  CAS  Google Scholar 

  • Desviat LD, Perez B, Belanger-Quintana A, Castro M, Aguado C, Sanchez A, Garcia MJ, Martinez-Pardo P, Ugarte M (2004) Tetrahydrobiopterin responsiveness: results of the BH4 loading test in 31 Spanish PKU patients and correlation with their genotype. Mol Genet Metab 83(1–2):157–162

    Article  PubMed  CAS  Google Scholar 

  • DiLella AG, Kwok SC, Ledley FD, Marvit J, Woo SL (1986) Molecular structure and polymorphic map of the human phenylalanine hydroxylase gene. Biochemistry 25(4):743–749

    Article  PubMed  CAS  Google Scholar 

  • Dipple KM, McCabe ER (2000) Modifier genes convert “simple” Mendelian disorders to complex traits. Mol Genet Metab 71(1–2):43–50

    Article  PubMed  CAS  Google Scholar 

  • Eiken HG, Knappskog PM, Boman H, Thune KS, Kaada G, Motzfeld K, Apold J (1996) Relative frequency, heterogeneity and geographic clustering of PKU mutations in Norway. Eur J Hum Genet 4(4):205–213

    PubMed  CAS  Google Scholar 

  • Erlandsen H, Stevens RC (2001) A structural hypothesis for BH4 responsiveness in patients with mild forms of hyperphenylananinaemia and phenylketonuria. J Inherit Metab Dis 24(2):213–230

    Article  PubMed  CAS  Google Scholar 

  • Fiori L, Fiege B, Riva E, Giovannini M (2005) Incidence of BH4-responsiveness in phenylalanine-hydroxylase-deficient Italian patients. Mol Genet Metab 86(suppl 1):67–74

    Article  Google Scholar 

  • Gersting SW, Kemter KF, Staudigl M, Messing DD, Danecke MK, Lagler FB, Sommerhoff CP, Roscher AA, Muntau AC (2008) Loss of function in phenylketonuria is caused by impaired molecular motions and conformational instability. Am J Hum Genet 83(1):5–17

    Article  PubMed  CAS  Google Scholar 

  • Gersting SW, Lagler FB, Eichinger A, Kemter KF, Danecka MK, Messing DD, Staudigl M, Domdey KA, Zsifkovits C, Fingerhut R, Glossmann H, Roscher AA, Muntau AC (2010) Pahenu1 is a mouse model for tetrahydrobiopterin-responsive phenylalanine hydroxylase deficiency and promotes analysis of the pharmacological chaperone mechanism in vivo. Hum Mol Genet 19(10):2039–2049

    Article  PubMed  CAS  Google Scholar 

  • Guldberg P, Romano V, Ceratto N, Bosco P, Ciuna M, Indelicato A, Mollica F, Meli C, Giovannini M, Riva E, Güttler F (1993) Mutational spectrum of phenylalanine hydroxylase deficiency in Sicily: implications for diagnosis of hyperphenylalaninemia in southern Europe. Hum Mol Genet 2(10):1703–1707

    Article  PubMed  CAS  Google Scholar 

  • Gulderberg P, Rey F, Zschocke J, Romano V, Francois B, Michiels L, Ullrich K, Hoffmann GF, Burgard P, Schmidt H, Meli C, Riva E, Dianzani I, Ponzone A, Rey J, Güttler F (1998) A European multicenter study of phenylalanine hydroxylase deficiency: classification of 105 mutations and a general system for genotype- based prediction of metabolic phenotype. Am J Hum Genet 63(1):71–79

    Article  Google Scholar 

  • Jaruzelska J, Matuszak R, Lyonnet S, Rey F, Rey J, Filipowicz J, Borski K, Munnich A (1993) Genetic background of clinical homogeneity of phenylketonuria in Poland. J Med Genet 30(3):232–234

    Article  PubMed  CAS  Google Scholar 

  • Kadasi L, Polakova H, Ferakova E, Hudecova S, Bohusova T, Szomolayova I, Strnova J, Hruskovic I, Moschonas NK, Ferak V (1995) PKU in Slovakia: mutation screening and haplotype analysis. Hum Genet 95(1):112–114

    Article  PubMed  CAS  Google Scholar 

  • Karacic I, Meili D, Sarnavka V, Heintz C, Thöny B, Ramadza DP, Fumic K, Mardesic D, Baric I, Blau N (2009) Genotype-predicted tetrahydrobiopterin (BH4)-responsiveness and molecular genetics in Croatian patients with Phenylalanine Hydroxylase (PAH) deficiency. Mol Genet Metab 97(3):165–171

    Article  PubMed  CAS  Google Scholar 

  • Kasper DC, Ratschmann R, Metz TF, Mechtler TP, Möslinger D, Konstantopoulou V, Item CB, Pollak A, Herkner KR (2010) The national Austrian Newborn Screening Program- eight years experience with mass spectrometry. Past, present, and future goals. Wien Klin Wochenschr 122(21–22):607–613

    Article  PubMed  Google Scholar 

  • Kayaalp E, Treacy E, Waters PJ, Byck S, Nowacki P, Scriver CR (1997) Human phenylalanine hydroxylase mutations and hyperphenylalaninaemia phenotypes: a metanalysis of genotype-phenotype correlations. Am J Hum Genet 61(6):1309–1317

    Article  PubMed  CAS  Google Scholar 

  • Kozak L, Blazkova M, Kuhrova V, Pijackova A, Ruzickova S, Stastna S (1997) Mutation and haplotype analysis of phenylalanine hydroxylase alleles in classical PKU patients from Czech Republic: identification of four novel mutations. J Med Genet 34(11):893–898

    Article  PubMed  CAS  Google Scholar 

  • Kozak L, Hrabincova E, Kintr J, Horky O, Zapletalova P, Blahakova I, Mejstrik P, Prochazkova D (2006) Identification and characterization of large deletions in the phenylalanine hydroxylase (PAH) gene by MLPA: evidence for both homologous and non-homologous mechanisms of rearrangement. Mol Genet Metab 89(4):300–309

    Article  PubMed  CAS  Google Scholar 

  • Kure S, Hou DC, Ohura T, Iwamoto H, Suzuki S, Sugiyama N, Sakamoto O, Fujii K, Matsubara Y, Narisawa K (1999) Tetrahydrobiopterin-responsive phenylalanine hydroxylase deficiency. J Pediatr 135(3):375–378

    Article  PubMed  CAS  Google Scholar 

  • Lagler FB, Gersting SW, Zsifkovits C, Steinbacher A, Eichinger A, Danecka MK, Staudigl M, Fingerhut R, Glossmann H, Muntau AC (2010) New insights into tetrahydrobiopterin pharmacodynamics from Pah enu1/2, a mouse model for compound heterozygous tetrahydrobiopterin-responsive phenylalanine hydroxylase deficiency. Biochem Pharmacol 80(10):1563–1571

    Article  PubMed  CAS  Google Scholar 

  • Leandro J, Nascimento C, de Almeida IT, Leandro P (2006) Co-expression of different subunits of human phenylalanine hydroxylase: evidence of negative interallelic complementation. Biochim Biophys Acta 1762(5):544–550

    Article  PubMed  CAS  Google Scholar 

  • Lilleväli H, Ounap K, Metspalu A (1996) Phenylalanine hydroxylase gene mutation R408W is present on 84 % of Estonian phenylketonuria chromosomes. Eur J Hum Genet 4(5):296–300

    PubMed  Google Scholar 

  • Mitchell JJ, Wilcken B, Alexander I, Ellaway C, O’Grady H, Wiley V, Earl J, Christodoulou J (2005) Tetrahydrobiopterin-responsive phenylketonuria: the New South Wales experience. Mol Genet Metab 86(suppl1):81–85

    Article  Google Scholar 

  • Møller LB, Nygren AO, Scott P, Hougaard P, Nielsen JB, Hartmann C, Güttler F, Tyfield L, Zschocke J (2007) Low proportion of whole exon deletions causing phenylketonuria in Denmark and Germany. Hum Mutat 28(2):207

    Article  Google Scholar 

  • Munk-Martin TL, Hyland K (2001) Tetrahydrobiopterin as a regulator of aromatic amino acid hydroxylase gene expression. Pteridines 12:65

    Google Scholar 

  • Pey AL, Desviat LR, Gamez A, Ugarte M, Perez B (2003) Phenylketonuria: genotype-phentotype correlations based on expression analysis of structural and functional mutations in PAH. Hum Mutat 21(4):370–378

    Article  PubMed  CAS  Google Scholar 

  • Pey AL, Perez B, Desviat LR, Martinez MA, Aguado C, Erlandsen H, Gamez A, Stevens RC, Thorolfsson M, Ugarte M, Martinez A (2004) Mechanisms underlying responsiveness to tetrahydrobiopterin in mild phenylketonuria mutations. Hum Mutat 24(5):388–399

    Article  PubMed  CAS  Google Scholar 

  • Pey AL, Stricher F, Serrano L, Martinez A (2007) Predicted effects of missense mutations on native-state stability account for phenotypic outcome in phenylketonuria, a paradigm of misfolding diseases. Am J Hum Genet 81(5):1006–1024

    Article  PubMed  CAS  Google Scholar 

  • Pronina N, Giannattasio S, Lattanzio P, Lugovska R, Vevere P, Kornejeva A (2003) The molecular basis of phenylketonuria in Latvia. Hum Mutat 21(4):398–399

    Article  PubMed  CAS  Google Scholar 

  • Scriver CR (2007) The PAH gene, phenylketonuria and a paradigm shift. Hum Mutat 28(9):831–845

    Article  PubMed  CAS  Google Scholar 

  • Scriver CR, Hurtubise M, Konecki D, Phommarinh M, Prevost L, Erlandsen H, Stevens R, Waters PJ, Ryan S, McDonald D, Sarkissian C (2003) PAHdb 2003: what a locus-specific knowledgebase can do. Hum Mutat 21(4):333–344

    Article  PubMed  CAS  Google Scholar 

  • Staudigl M, Gersting SW, Danecka MK, Messing DD, Woidy M, Pinkas D, Kemter KF, Blau N, Muntau AC (2011) The interplay between genotype, metabolic state and cofactor treatment governs phenylalanine hydroxylase function and drug response. Hum Mol Genet 20(13):2628–2641

    Article  PubMed  CAS  Google Scholar 

  • Trefz FK, Scheible D, Götz H, Frauendienst-Egger G (2009) Significance of genotype in tetrahydrobiopterin-responsive phenylketonuria. J Inherit Metab Dis 32(1):22–26

    Article  PubMed  CAS  Google Scholar 

  • Zschocke J (2003) Phenylketonuria mutations in Europe. Hum Mutat 21(4):345–356

    Article  PubMed  CAS  Google Scholar 

  • Zschocke J, Hoffman GF (1999) Phenylketonuria mutations in Germany. Hum Genet 104(5):390–398

    Article  PubMed  CAS  Google Scholar 

  • Zschocke J, Quak E, Guldberg P, Hofmann GF (2000) Mutation analysis in glutaric aciduria type I. J Med Genet 37(3):177–181

    Article  PubMed  CAS  Google Scholar 

  • Zurflüh MR, Zschocke J, Lindner M, Feillet F, Chery C, Burlina A, Stevens RC, Thöny B, Blau N (2008) Molecular genetics of tetrahydrobiopterin-responsive phenylalanine hydroxylase deficiency. Hum Mutat 29(1):167–175

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

We want to thank Martina Judmaier and Bernadette Knafl, dietician, Medical University Graz and Anna Fekete, dietician, Medical University Vienna for their assistance with dietary information and newborn levels of phenylalanine.

This work has been partly funded by Invita “Gesellschaft zur Förderung der Gesundheit unserer Kinder”.

Conflict of interest

None.

Author information

Authors and Affiliations

  1. Department of Pediatric and Adolescent Surgery, Medical University of Graz, Graz, Austria

    Elisabeth Sterl

  2. Department of Pediatrics, Medical University of Graz, Graz, Austria

    Karl Paul, Eduard Paschke, Michaela Brunner-Krainz, Eva Windisch & Barbara Plecko

  3. Division of Human Genetics, Medical University Innsbruck, Innsbruck, Austria

    Johannes Zschocke

  4. Department of Pediatrics, Medical University of Vienna, Vienna, Austria

    Vassiliki Konstantopoulou & Dorothea Möslinger

  5. Department of Pediatrics, Medical University of Innsbruck, Innsbruck, Austria

    Daniela Karall & Sabine Scholl-Bürgi

  6. Department of Pediatrics, Paracelsus Medical University, Salzburg, Austria

    Wolfgang Sperl & Florian Lagler

  7. Institute for Inborn Errors of Metabolism, Paracelsus Medical University, Salzburg, Austria

    Florian Lagler

  8. Department of Pediatrics, University of Zürich, Zürich, Switzerland

    Barbara Plecko

Authors
  1. Elisabeth Sterl
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Karl Paul
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eduard Paschke
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Johannes Zschocke
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Michaela Brunner-Krainz
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Eva Windisch
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Vassiliki Konstantopoulou
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Dorothea Möslinger
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Daniela Karall
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Sabine Scholl-Bürgi
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Wolfgang Sperl
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Florian Lagler
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Barbara Plecko
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Barbara Plecko.

Additional information

Communicated by: Nenad Blau

Electronic supplementary material

Below is the link to the electronic supplementary material.

Suppl. Table 1

Disease causing mutations of the PAH Gene (GenBank U49897.1, NM_000277) and information on phenotype as well as newborn phenylalanine (phe) levels of 147 unrelated Austrian patients with PAH deficiency (PKU; MIM 261600) included in this study (DOC 302 kb)

Suppl. Table 2

Mutations with an allele frequency <1.5% within our cohort of 147 Austrian PAH deficiency Patients (PKU; MIM 261600), number of affected alleles and population background. Number of affected alleles in case of different population background are enclosed in brackets (DOC 70 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sterl, E., Paul, K., Paschke, E. et al. Prevalence of tetrahydrobiopterine (BH4)-responsive alleles among Austrian patients with PAH deficiency: comprehensive results from molecular analysis in 147 patients. J Inherit Metab Dis 36, 7–13 (2013). https://doi.org/10.1007/s10545-012-9485-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10545-012-9485-y

Keywords

Search

Navigation