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JIMD Reports, Volume 37 pp 73–83Cite as

Hyperphenylalaninemia Correlated with Global Decrease of Antioxidant Genes Expression in White Blood Cells of Adult Patients with Phenylketonuria

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Part of the book series: JIMD Reports ((JIMD,volume 37))

Abstract

Background: Several studies have highlighted disturbance of redox homeostasis in patients with phenylketonuria (PKU) which may be associated with neurological disorders observed in patients, especially during adulthood when phenylalanine restrictive diets are not maintained. The aim of this study was to assess the antioxidant profile in a cohort of PKU patients in comparison to the controls and to evaluate its relation to biochemical parameters especially phenylalaninemia.

Methods: We measured RNA expression of 22 antioxidant genes and reactive oxygen species (ROS) levels in white blood cells of 10 PKU patients and 10 age- and gender-matched controls. We also assessed plasma amino acids, vitamins, oligo-elements, and urinary organic acids concentrations. Then we evaluated the relationship between redox status and biochemical parameters.

Results: In addition to expected biochemical disturbances, we highlighted a significant global decrease of antioxidant genes expression in PKU patients in comparison to the controls. This global decrease of antioxidant genes expression, including various isoforms of peroxiredoxins, glutaredoxins, glutathione peroxidases, and superoxide dismutases, was significantly correlated to hyperphenylalaninemia.

Conclusion: This study is the first to evaluate the expression of 22 antioxidant genes in white blood cells regarding biochemical parameters in PKU. These findings highlight the association of hyperphenylalaninemia with antioxidant genes expression. New experiments to specify the role of oxidative stress in PKU pathogenesis may be useful in suggesting new recommendations in PKU management and new therapeutic trials based on antioxidant defenses.

The authors “Olivier Herault” and “François Maillot” contributed equally.

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Abbreviations

BMI:

Body mass index

CAT:

Catalase

Ct:

Cycle threshold

GLRX:

Glutaredoxin

GPX:

Glutathione peroxidase

GSR:

Glutathione-disulfide reductase

Phe:

Phenylalanine

PKU:

Phenylketonuria

PRDX:

Peroxiredoxin

ROS:

Reactive oxygen species

RQ:

Relative quantification

SOD:

Superoxide dismutase

TXN:

Thioredoxin

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Acknowledgements

We thank the SFEIM (Société Française des Erreurs Innées du Métabolisme) for its support. We also thank Penelope HODGES for her participation in PKU patients’ recruitment and for the English proofreading.

Author information

Authors and Affiliations

  1. Laboratoire de biochimie et biologie moléculaire, hôpital Bretonneau CHRU de Tours, Tours, France

    Charlotte Veyrat-Durebex, Hélène Blasco, Franck Patin & Catherine Antar

  2. INSERM U930, Université François Rabelais Tours, Tours, France

    Charlotte Veyrat-Durebex, Hélène Blasco, Franck Patin, Patrick Emond & Catherine Antar

  3. CNRS UMR 7292 - équipe LNOx, Université François Rabelais Tours, Tours, France

    Christelle Debeissat & Olivier Herault

  4. Service d’hématologie biologique, CHRU de Tours, Tours, France

    Christelle Debeissat & Olivier Herault

  5. INSERM CIC 1415, CHRU de Tours, Tours, France

    Valérie Gissot

  6. Service de médecine interne, CHRU de Tours, Tours, France

    Hélène Henique & François Maillot

  7. INSERM U1069, Université François Rabelais Tours, Tours, France

    Hélène Henique & François Maillot

Authors
  1. Charlotte Veyrat-Durebex
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  2. Christelle Debeissat
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  3. Hélène Blasco
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  4. Franck Patin
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  5. Hélène Henique
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  7. Catherine Antar
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  8. Valérie Gissot
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  9. Olivier Herault
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  10. François Maillot
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Corresponding author

Correspondence to Charlotte Veyrat-Durebex .

Editor information

Editors and Affiliations

  1. Tulane University Medical School , New Orleans, Louisiana, USA

    Eva Morava

  2. Division of Metabolism & Children's Research Centre, University Children's Hospital Zürich, Zürich, Switzerland

    Matthias Baumgartner

  3. Division of Child and Adolescent Neurology, Mayo Clinic, Rochester, Minnesota, USA

    Marc Patterson

  4. Clinical and Molecular Genetics Unit, UCL Institute of Child Health, London, United Kingdom

    Shamima Rahman

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

    Johannes Zschocke

  6. Center for Child and Adolescent Medicine, Heidelberg University Hospital, Heidelberg, Germany

    Verena Peters

Additional information

Communicated by: Nenad Blau, PhD

Take-Home Message (Synopsis)

This preliminary study highlights a lower antioxidant arsenal in leukocytes associated with hyperphenylalaninemia using assessment of gene expression of 22 major antioxidant enzymes in adult PKU patients.

Conflict of Interest

All authors declare that they have no conflicts of interest with the contents of this chapter.

Informed Consent

All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000. Informed consent was obtained from all patients for being included in the study. Proof that informed consent was obtained are available upon request.

Author Contributions

C.V-D designed and performed experiments, analyzed data, and wrote the manuscript; C.D performed and supervised redox status experiments and gave technical and intellectual support on redox study; H.B designed experiments, supervised the writing of the manuscript, and gave intellectual support and conceptual advice; F.P performed experiments; H.H collected clinical data; P.E supervised biochemical parameters assays; C.A performed biochemical experiments; V.G recruited healthy controls and participated in the design of study; O.H gave technical and intellectual support on redox status interpretation and supervised the writing of the manuscript; F.M designed and supervised the study, recruited PKU patients, and supervised the writing of the manuscript. All authors provided conceptual advices and comments on the manuscript.

Author who serves as guarantor: François Maillot.

Details of Funding

This study was funded by a research grant from the University Hospital of Tours. The authors confirm independence from the sponsors; the content of the chapter has not been influenced by the sponsors.

Details of Ethics Approval

This study was approved by the ethics committee of University Hospital of Tours (CPPRB, for Comité de Protection des Personnes) and registered on ClinicalTrials.gov (study ID number PHAO14/FM-StressOX-PCU, “Antioxidant Signature in Adults Patients with Phenlyketonuria”). Informed consent was signed by all patients and controls prior to inclusion.

Supplementary Material

Fig. S1

Profile of expression of antioxidant genes of different subcategories of PKU patients compared to healthy controls (fold change ratio of gene expression in PKU patients compared to controls). This graph represented antioxidant profile of 3 particular patients (curves represent expression of the 26 studied transcripts in patient compared to their matched controls): a patient receiving BH4 treatment (excluded from statistical analysis, red curve), a patient with permanent moderate hyperphenylalaninemia (yellow curve), and a patient with atypical PKU (green curve). Antioxidant profiles of these 3 patients were presented in comparison to the mean antioxidant profile of 7 typical PKU patients (black curve). RNA isolated from white blood cells of PKU patients and controls was reverse transcribed, and a real-time PCR using a custom panel was performed on cDNA in order to assess the expression of main antioxidant genes. Results of real-time PCR assays are presented as fold change of genes expression in white blood cells of PKU patients compared to controls. Genes of the antioxidant profile are presented in order from most to least expressed in X-axis, according to their ΔCt values (versus mean references) (JPEG 99 kb)

Table S1

Patients’ characteristics (DOCX 16 kb)

Table S2

Comparison of biochemical parameters concentrations in plasma of PKU patients and controls (DOCX 17 kb)

Table S3

Comparison of urinary concentrations of organic acids of PKU patients and controls. Levels of urinary concentrations of organic acids are given in arbitrary unit of peak area in chromatogram, reported to area peak of internal standard and reported to urinary concentration of creatinine (DOCX 18 kb)

Supplementary Methods (DOCX 15 kb)

Supplementary Data (XLSX 26 kb)

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Veyrat-Durebex, C. et al. (2017). Hyperphenylalaninemia Correlated with Global Decrease of Antioxidant Genes Expression in White Blood Cells of Adult Patients with Phenylketonuria. In: Morava, E., Baumgartner, M., Patterson, M., Rahman, S., Zschocke, J., Peters, V. (eds) JIMD Reports, Volume 37. JIMD Reports, vol 37. Springer, Berlin, Heidelberg. https://doi.org/10.1007/8904_2017_16

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  • DOI: https://doi.org/10.1007/8904_2017_16

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