title: Molecular basis of pyruvate kinase deficiency among Tunisians: description of new mutations affecting coding and noncoding regions in the PKLR gene.
abstract: BACKGROUND
  Pyruvate kinase (PK) deficiency is one of the most common hereditary nonspherocytic hemolytic anemias worldwide with clinical manifestations ranging from mild to severe hemolysis. However, investigation of this enzymopathy is lacking in Tunisia. We report here a pioneer investigation of PK deficiency among Tunisian cases referred to our laboratory for biological analysis of unknown cause of hemolytic anemia.
  
  
  METHODS
  Two hundred and fifty-three patients with unknown cause of hemolytic anemia have been addressed to our laboratory in order to investigate for red blood cells genetic disorders. Red cell enzyme activities were measured by standard methods, and molecular analysis was performed by DNA sequencing. The interpretation of mutation effect and the molecular modeling were performed by using specific software.
  
  
  RESULTS
  Six different PKLR mutations were found (c.966-1G>T; c.965+1G>A; c.721G>T; c.1163C>A; c.1456C>T; c.1537T>A), among which four are described for the first time. Genotype-phenotype correlations for the novel missense mutations were investigated by three-dimensional structure analysis.
  
  
  CONCLUSIONS
  This study provides important data of PK deficiency among Tunisians. It might be followed by a large neonatal screening to determine the spectrum of PK mutations and identify potential deficient patients for an early medical follow-up.
authors.affiliation: Laboratoire d'Hématologie Moléculaire et Cellulaire, Institut Pasteur de Tunis, Université de Tunis El Manar, Tunis, Tunisia.
authors.forename: M
authors.initials: M
authors.lastname: Jaouani
authors.affiliation: Unidade de Hematlogia Molecular, Centro Hospitalar e Universitário de Coimbra (CHUC), Coimbra, Portugal.|Research Centre for Anthropology and Health (CIAS), Department of Life Sciences, Universidade de Coimbra, Coimbra, Portugal.
authors.forename: L
authors.initials: L
authors.lastname: Manco
authors.affiliation: Laboratoire d'Hématologie Moléculaire et Cellulaire, Institut Pasteur de Tunis, Université de Tunis El Manar, Tunis, Tunisia.
authors.forename: M
authors.initials: M
authors.lastname: Kalai
authors.affiliation: Laboratoire d'Hématologie Moléculaire et Cellulaire, Institut Pasteur de Tunis, Université de Tunis El Manar, Tunis, Tunisia.
authors.forename: L
authors.initials: L
authors.lastname: Chaouch
authors.affiliation: Laboratoire d'Hématologie Moléculaire et Cellulaire, Institut Pasteur de Tunis, Université de Tunis El Manar, Tunis, Tunisia.
authors.forename: K
authors.initials: K
authors.lastname: Douzi
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.
authors.forename: A
authors.initials: A
authors.lastname: Silva
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.
authors.forename: S
authors.initials: S
authors.lastname: Macedo
authors.affiliation: Laboratoire d'Hématologie Moléculaire et Cellulaire, Institut Pasteur de Tunis, Université de Tunis El Manar, Tunis, Tunisia.
authors.forename: I
authors.initials: I
authors.lastname: Darragi
authors.affiliation: Laboratoire d'Hématologie Moléculaire et Cellulaire, Institut Pasteur de Tunis, Université de Tunis El Manar, Tunis, Tunisia.
authors.forename: I
authors.initials: I
authors.lastname: Boudriga
authors.affiliation: Laboratoire d'Hématologie Moléculaire et Cellulaire, Institut Pasteur de Tunis, Université de Tunis El Manar, Tunis, Tunisia.
authors.forename: D
authors.initials: D
authors.lastname: Chaouachi
authors.affiliation: Service de pédiatrie-urgences-consultations, Hôpital d'Enfants de Tunis, Tunis, Tunisia.
authors.forename: Z
authors.initials: Z
authors.lastname: Fitouri
authors.affiliation: Laboratory for Red Blood Cell Research, Department of Clinical Chemistry and Hematology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: R
authors.initials: R
authors.lastname: Van Wijk
authors.affiliation: Unidade de Hematlogia Molecular, Centro Hospitalar e Universitário de Coimbra (CHUC), Coimbra, Portugal.
authors.forename: M L
authors.initials: ML
authors.lastname: Ribeiro
authors.affiliation: Laboratoire d'Hématologie Moléculaire et Cellulaire, Institut Pasteur de Tunis, Université de Tunis El Manar, Tunis, Tunisia.
authors.forename: S
authors.initials: S
authors.lastname: Abbes
mesh_terms: D000743:Anemia, Hemolytic / Q000209:etiology / Q000235:genetics; D000746:Anemia, Hemolytic, Congenital Nonspherocytic / Q000209:etiology / Q000235:genetics*; D004252:DNA Mutational Analysis; D004912:Erythrocytes / Q000201:enzymology; D056726:Genetic Association Studies; D006801:Humans; D008958:Models, Molecular; D020125:Mutation, Missense / Q000235:genetics*; D011770:Pyruvate Kinase / Q000172:deficiency* / Q000235:genetics; D015323:Pyruvate Metabolism, Inborn Errors / Q000209:etiology / Q000235:genetics*; D014416:Tunisia / Q000453:epidemiology
chemical_list: D011770:Pyruvate Kinase
keywords: PKLR mutations; Pyruvate kinase deficiency; Tunisian families; genotype-phenotype correlation; hemolytic anemia