title: Therapy of Fabry disease with pharmacological chaperones: from in silico predictions to in vitro tests.
abstract: BACKGROUND
  Fabry disease is a rare disorder caused by a large variety of mutations in the gene encoding lysosomal alpha-galactosidase. Many of these mutations are unique to individual families. Fabry disease can be treated with enzyme replacement therapy, but a promising novel strategy relies on small molecules, so called "pharmacological chaperones", which can be administered orally. Unfortunately only 42% of genotypes respond to pharmacological chaperones.
  
  
  RESULTS
  A procedure to predict which genotypes responsive to pharmacological chaperones in Fabry disease has been recently proposed. The method uses a position-specific substitution matrix to score the mutations. Using this method, we have screened public databases for predictable responsive cases and selected nine representative mutations as yet untested with pharmacological chaperones. Mutant lysosomal alpha galactosidases were produced by site directed mutagenesis and expressed in mammalian cells. Seven out of nine mutations responded to pharmacological chaperones. Nineteen other mutations that were tested with pharmacological chaperones, but were not included in the training of the predictive method, were gathered from literature and analyzed in silico. In this set all five mutations predicted to be positive were responsive to pharmacological chaperones, bringing the percentage of responsive mutations among those predicted to be positive and not used to train the classifier to 86% (12/14). This figure differs significantly from the percentage of responsive cases observed among all the Fabry mutants tested so far.
  
  
  CONCLUSIONS
  In this paper we provide experimental support to an "in silico" method designed to predict missense mutations in the gene encoding lysosomal alpha galactosidase responsive to pharmacological chaperones. We demonstrated that responsive mutations can be predicted with a low percentage of false positive cases. Most of the mutations tested to validate the method were described in the literature as associated to classic or mild classic phenotype. The analysis can provide a guideline for the therapy with pharmacological chaperones supported by experimental results obtained in vitro. We are aware that our results were obtained in vitro and cannot be translated straightforwardly into benefit for patients, but need to be validated by clinical trials.
authors: Istituto di Chimica Biomolecolare - CNR, Pozzuoli, Italy. giuseppina.andreotti@icb.cnr.it
authors: Giuseppina
authors: G
authors: Andreotti
authors: Valentina
authors: V
authors: Citro
authors: Agostina
authors: A
authors: De Crescenzo
authors: Pierangelo
authors: P
authors: Orlando
authors: Marco
authors: M
authors: Cammisa
authors: Antonella
authors: A
authors: Correra
authors: Maria Vittoria
authors: MV
authors: Cubellis
mesh_terms: D017485:1-Deoxynojirimycin / Q000494:pharmacology* / Q000627:therapeutic use; D000818:Animals; D001483:Base Sequence; D019556:COS Cells / Q000378:metabolism; D020134:Catalytic Domain; D002522:Chlorocebus aethiops; D000795:Fabry Disease / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000494:pharmacology* / Q000627:therapeutic use; D008969:Molecular Sequence Data; D009113:Muramidase; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D011237:Predictive Value of Tests; D000519:alpha-Galactosidase / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism
chemical_list: D018832:Molecular Chaperones; D017485:1-Deoxynojirimycin; D009113:Muramidase; D000519:alpha-Galactosidase