{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,6]],"date-time":"2026-02-06T22:22:18Z","timestamp":1770416538711,"version":"3.49.0"},"reference-count":49,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2017,9,24]],"date-time":"2017-09-24T00:00:00Z","timestamp":1506211200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Background: Rare diseases represent a challenge for physicians because patients are rarely seen, and they can manifest with symptoms similar to those of common diseases. In this work, genetic confirmation of diagnosis is derived from DNA sequencing. We present a tutorial for the molecular analysis of a rare disease using Fabry disease as an example. Methods: An exonic sequence derived from a hypothetical male patient was matched against human reference data using a genome browser. The missense mutation was identified by running BlastX, and information on the affected protein was retrieved from the database UniProt. The pathogenic nature of the mutation was assessed with PolyPhen-2. Disease-specific databases were used to assess whether the missense mutation led to a severe phenotype, and whether pharmacological therapy was an option. Results: An inexpensive bioinformatics approach is presented to get the reader acquainted with the diagnosis of Fabry disease. The reader is introduced to the field of pharmacological chaperones, a therapeutic approach that can be applied only to certain Fabry genotypes. Conclusion: The principle underlying the analysis of exome sequencing can be explained in simple terms using web applications and databases which facilitate diagnosis and therapeutic choices.<\/jats:p>","DOI":"10.3390\/ijms18102049","type":"journal-article","created":{"date-parts":[[2017,9,26]],"date-time":"2017-09-26T04:28:01Z","timestamp":1506400081000},"page":"2049","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["E-Learning for Rare Diseases: An Example Using Fabry Disease"],"prefix":"10.3390","volume":"18","author":[{"given":"Chiara","family":"Cimmaruta","sequence":"first","affiliation":[{"name":"Dipartimento di Biologia, Universit\u00e0 Federico II, 80126 Napoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ludovica","family":"Liguori","sequence":"additional","affiliation":[{"name":"Istituto di Chimica Biomolecolare, Consiglio Nazionale delle Ricerche, Comprensorio Olivetti, Edificio 70, Via Campi Flegrei 34, 80078 Pozzuoli, Italy"},{"name":"Dipartimento di Scienze e Tecnologie Ambientali, Biologiche e Farmaceutiche, Universit\u00e0 della Campania \u201cLuigi Vanvitelli\u201d, 81100 Caserta, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3136-2138","authenticated-orcid":false,"given":"Maria","family":"Monticelli","sequence":"additional","affiliation":[{"name":"Dipartimento di Biologia, Universit\u00e0 Federico II, 80126 Napoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1594-0156","authenticated-orcid":false,"given":"Giuseppina","family":"Andreotti","sequence":"additional","affiliation":[{"name":"Istituto di Chimica Biomolecolare, Consiglio Nazionale delle Ricerche, Comprensorio Olivetti, Edificio 70, Via Campi Flegrei 34, 80078 Pozzuoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Valentina","family":"Citro","sequence":"additional","affiliation":[{"name":"Dipartimento di Biologia, Universit\u00e0 Federico II, 80126 Napoli, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2017,9,24]]},"reference":[{"key":"ref_1","unstructured":"(2017, September 19). Orphanet. Available online: http:\/\/www.Orpha.Net\/consor\/cgi-bin\/index.Php."},{"key":"ref_2","unstructured":"Online Mendelian Inheritance in Man, O (2017, September 19). Mckusick-Nathans Institute of Genetic Medicine, Johns Hopkins University (Baltimore, MD). Available online: https:\/\/omim.Org\/."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1186\/1750-1172-5-30","article-title":"Fabry disease","volume":"5","author":"Germain","year":"2010","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1136\/hrt.2005.063818","article-title":"The heart in anderson-fabry disease and other lysosomal storage disorders","volume":"93","author":"Linhart","year":"2007","journal-title":"Heart"},{"key":"ref_5","unstructured":"Schiffmann, R., and Moore, D.F. (2006). Neurological manifestations of fabry disease. Fabry Disease: Perspectives from 5 Years of FOS, Oxford PharmaGenesis."},{"key":"ref_6","unstructured":"Sunder-Plassmann, G. (2006). Renal manifestations of fabry disease. Fabry Disease: Perspectives from 5 Years of FOS, Oxford PharmaGenesis."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Citro, V., Cammisa, M., Liguori, L., Cimmaruta, C., Lukas, J., Cubellis, M.V., and Andreotti, G. (2016). The large phenotypic spectrum of fabry disease requires graduated diagnosis and personalized therapy: A meta-analysis can help to differentiate missense mutations. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17122010"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1086\/504601","article-title":"High incidence of later-onset fabry disease revealed by newborn screening","volume":"79","author":"Spada","year":"2006","journal-title":"Am. J. Hum. Genet."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1016\/j.jpeds.2013.01.031","article-title":"Identification of infants at risk for developing Fabry, Pompe, or mucopolysaccharidosis-I from newborn blood spots by tandem mass spectrometry","volume":"163","author":"Scott","year":"2013","journal-title":"J. Pediatr."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1016\/j.jpeds.2014.09.023","article-title":"Lysosomal storage disorder screening implementation: Findings from the first six months of full population pilot testing in missouri","volume":"166","author":"Hopkins","year":"2015","journal-title":"J. Pediatr."},{"key":"ref_11","unstructured":"(2017, September 19). HGMD. Available online: http:\/\/www.Hgmd.Cf.Ac.Uk\/."},{"key":"ref_12","first-page":"855","article-title":"High frequency of the gaucher disease mutation at nucleotide 1226 among ashkenazi jews","volume":"49","author":"Zimran","year":"1991","journal-title":"Am. J. Hum. Genet."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1186\/s13023-015-0253-6","article-title":"Recommendations for initiation and cessation of enzyme replacement therapy in patients with fabry disease: The european fabry working group consensus document","volume":"10","author":"Biegstraaten","year":"2015","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1547","DOI":"10.1681\/ASN.2006080816","article-title":"Sustained, long-term renal stabilization after 54 months of agalsidase beta therapy in patients with fabry disease","volume":"18","author":"Germain","year":"2007","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1186\/1750-1172-8-47","article-title":"Long term enzyme replacement therapy for fabry disease: Effectiveness on kidney, heart and brain","volume":"8","author":"Rombach","year":"2013","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1111\/joim.12077","article-title":"Long-term outcome of enzyme-replacement therapy in advanced fabry disease: Evidence for disease progression towards serious complications","volume":"274","author":"Weidemann","year":"2013","journal-title":"J. Intern. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1056\/NEJMoa1510198","article-title":"Treatment of fabry\u2019s disease with the pharmacologic chaperone migalastat","volume":"375","author":"Germain","year":"2016","journal-title":"N. Engl. J. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1136\/jmedgenet-2016-104178","article-title":"Oral pharmacological chaperone migalastat compared with enzyme replacement therapy in fabry disease: 18-month results from the randomised phase III attract study","volume":"54","author":"Hughes","year":"2016","journal-title":"J. Med. Genet."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1147","DOI":"10.1007\/s40265-016-0607-y","article-title":"Migalastat: First global approval","volume":"76","author":"Markham","year":"2016","journal-title":"Drugs"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1186\/s13023-015-0273-2","article-title":"Drug repositioning can accelerate discovery of pharmacological chaperones","volume":"10","author":"Citro","year":"2015","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1097\/00042560-199510050-00008","article-title":"The tolerability and pharmacokinetics of N-butyl-deoxynojirimycin in patients with advanced hiv disease (ACTG 100). The aids clinical trials group (ACTG) of the national institute of allergy and infectious diseases","volume":"10","author":"Tierney","year":"1995","journal-title":"J. Acquir. Immune Defic. Syndr. Hum. Retrovirol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1517\/14656566.8.4.427","article-title":"Pharmacotherapeutic strategies using small molecules for the treatment of glycolipid lysosomal storage disorders","volume":"8","author":"Butters","year":"2007","journal-title":"Expert Opin. Pharmacother."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4816","DOI":"10.1021\/bi9002265","article-title":"Effects of pH and iminosugar pharmacological chaperones on lysosomal glycosidase structure and stability","volume":"48","author":"Lieberman","year":"2009","journal-title":"Biochemistry"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1513","DOI":"10.4155\/fmc.11.117","article-title":"Iminosugars as therapeutic agents: Recent advances and promising trends","volume":"3","author":"Nash","year":"2011","journal-title":"Future Med. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1038\/4801","article-title":"Accelerated transport and maturation of lysosomal alpha-galactosidase a in fabry lymphoblasts by an enzyme inhibitor","volume":"5","author":"Fan","year":"1999","journal-title":"Nat. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1038\/nature19057","article-title":"Analysis of protein-coding genetic variation in 60,706 humans","volume":"536","author":"Lek","year":"2016","journal-title":"Nature"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"D862","DOI":"10.1093\/nar\/gkv1222","article-title":"Clinvar: Public archive of interpretations of clinically relevant variants","volume":"44","author":"Landrum","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1038\/nmeth0410-248","article-title":"A method and server for predicting damaging missense mutations","volume":"7","author":"Adzhubei","year":"2010","journal-title":"Nat. Methods"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"D668","DOI":"10.1093\/nar\/gkj067","article-title":"Drugbank: A comprehensive resource for in silico drug discovery and exploration","volume":"34","author":"Wishart","year":"2006","journal-title":"Nucleic Acids Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1186\/1750-1172-5-36","article-title":"Prediction of the responsiveness to pharmacological chaperones: Lysosomal human alpha-galactosidase, a case of study","volume":"5","author":"Andreotti","year":"2010","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"638","DOI":"10.1039\/C6MB00866F","article-title":"Screening methods for identifying pharmacological chaperones","volume":"13","author":"Shin","year":"2017","journal-title":"Mol. Biosyst."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1038\/gim.2016.122","article-title":"The validation of pharmacogenetics for the identification of fabry patients to be treated with migalastat","volume":"19","author":"Benjamin","year":"2016","journal-title":"Genet. Med."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1214","DOI":"10.1016\/j.bbagen.2013.12.018","article-title":"A thermodynamic assay to test pharmacological chaperones for fabry disease","volume":"1840","author":"Andreotti","year":"2014","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1186\/1479-7364-2-5-297","article-title":"Fabry disease: Identification of 50 novel alpha-galactosidase a mutations causing the classic phenotype and three-dimensional structural analysis of 29 missense mutations","volume":"2","author":"Shabbeer","year":"2006","journal-title":"Hum. Genom."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1628","DOI":"10.1093\/ndt\/gfm813","article-title":"Agalsidase therapy in patients with fabry disease on renal replacement therapy: A nationwide study in italy","volume":"23","author":"Mignani","year":"2008","journal-title":"Nephrol. Dial. Transplant."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1016\/j.clinbiochem.2014.02.014","article-title":"Fabry disease: A new approach for the screening of females in high-risk groups","volume":"47","author":"Pasqualim","year":"2014","journal-title":"Clin. Biochem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1056\/NEJM199102073240607","article-title":"An atypical variant of fabry\u2019s disease with manifestations confined to the myocardium","volume":"324","author":"Eng","year":"1991","journal-title":"N. Engl. J. Med."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1181","DOI":"10.1038\/gim.2016.55","article-title":"Is it fabry disease?","volume":"18","author":"Schiffmann","year":"2016","journal-title":"Genet. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1186\/1750-1172-8-111","article-title":"Fabry_cep: A tool to identify fabry mutations responsive to pharmacological chaperones","volume":"8","author":"Cammisa","year":"2013","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1186\/1750-1172-6-66","article-title":"Therapy of fabry disease with pharmacological chaperones: From in silico predictions to in vitro tests","volume":"6","author":"Andreotti","year":"2011","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Lukas, J., Giese, A.K., Markoff, A., Grittner, U., Kolodny, E., Mascher, H., Lackner, K.J., Meyer, W., Wree, P., and Saviouk, V. (2013). Functional characterisation of alpha-galactosidase a mutations as a basis for a new classification system in fabry disease. PLoS Genet., 9.","DOI":"10.1371\/journal.pgen.1003632"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Cammisa, M., Correra, A., Andreotti, G., and Cubellis, M.V. (2013). Identification and analysis of conserved pockets on protein surfaces. BMC Bioinform., 14.","DOI":"10.1186\/1471-2105-14-S7-S9"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1138","DOI":"10.1038\/mt.2015.62","article-title":"Pharmacological chaperone therapy: Preclinical development, clinical translation, and prospects for the treatment of lysosomal storage disorders","volume":"23","author":"Parenti","year":"2015","journal-title":"Mol. Ther."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1220","DOI":"10.1002\/jnr.23755","article-title":"Chaperones as potential therapeutics for krabbe disease","volume":"94","author":"Graziano","year":"2016","journal-title":"J. Neurosci. Res."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1527","DOI":"10.2174\/1389450117666160813180812","article-title":"Therapeutic approaches using riboflavin in mitochondrial energy metabolism disorders","volume":"17","author":"Henriques","year":"2016","journal-title":"Curr. Drug Targets"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1515","DOI":"10.2174\/1389450117666160307143512","article-title":"Pharmacological chaperones that protect tetrahydrobiopterin dependent aromatic amino acid hydroxylases through different mechanisms","volume":"17","author":"Hole","year":"2016","journal-title":"Curr. Drug Targets"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"37583","DOI":"10.1038\/srep37583","article-title":"Identification of small molecule compounds for pharmacological chaperone therapy of aspartylglucosaminuria","volume":"6","author":"Banning","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1877","DOI":"10.1093\/hmg\/dds001","article-title":"Novel pharmacological chaperones that correct phenylketonuria in mice","volume":"21","author":"Kirchmair","year":"2012","journal-title":"Hum. Mol. Genet."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1038\/jhg.2011.31","article-title":"Fabry-database.Org: Database of the clinical phenotypes, genotypes and mutant alpha-galactosidase a structures in fabry disease","volume":"56","author":"Saito","year":"2011","journal-title":"J. Hum. Genet."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/18\/10\/2049\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:45:47Z","timestamp":1760208347000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/18\/10\/2049"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,9,24]]},"references-count":49,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2017,10]]}},"alternative-id":["ijms18102049"],"URL":"https:\/\/doi.org\/10.3390\/ijms18102049","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,9,24]]}}}