Therapy of Fabry disease with pharmacological chaperones: from in silico predictions to in vitro tests(1806 views) Andreotti G, Citro V, De Crescenzo A, Orlando P, Cammisa M, Correra A, Cubellis MV
Orphanet Journal Of Rare Diseases (ISSN: 1750-1172), 2011 Oct 17; 6: 66-66.
Keywords: 1-Deoxynojirimycin Pharmacology Therapeutic Use, Animals, Base Sequence, Cos Cells Metabolism, Catalytic Domain, Cercopithecus Aethiops, Fabry Disease Drug Therapy Enzymology Genetics, Female, Humans, Models, Molecular, Molecular Chaperones Pharmacology Therapeutic Use, Molecular Sequence Data, Muramidase, Mutagenesis, Site-Directed, Mutation, Missense, Predictive Value Of Tests, Alpha-Galactosidase Drug Effects Genetics Metabolism, Alpha Galactosidase, Migalastat, 1 Deoxynojirimycin, Lysozyme, Animal Cell, Article, Computer Model, Drug Mechanism, Gene Mutation, Genotype, In Vitro Study, Nonhuman, Western Blotting, Cell Strain Cos1, Chemical Structure, Enzyme Active Site, Evaluation, Missense Mutation, Molecular Genetics, Nucleotide Sequence, Site Directed Mutagenesis,
Affiliations: *** IBB - CNR ***
Istituto di Chimica Biomolecolare - CNR, Pozzuoli, Italy. giuseppina.andreotti@icb.cnr.it
Institute of Genetics and Biophysics A. Buzzati Traverso, CNR, Naples, Italy
Dipartimento di Scienze Ambientali, Seconda Università di Napoli, Caserta, Italy
Institute of Protein Biochemistry, CNR, Napoli, Italy
Dipartimento di Biologia Strutturale e Funzionale, Università Federico II, Napoli, Italy
Istituto di Biostrutture e Bioimmagini, CNR, Napoli, Italy
References: Not available.
Therapy of Fabry disease with pharmacological chaperones: from in silico predictions to in vitro tests
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.
Therapy of Fabry disease with pharmacological chaperones: from in silico predictions to in vitro tests