Clinical Epigenetics (Oct 2024)
Integrating D4Z4 methylation analysis into clinical practice: improvement of FSHD molecular diagnosis through distinct thresholds for 4qA/4qA and 4qA/4qB patients
- Claudia Strafella,
- Domenica Megalizzi,
- Giulia Trastulli,
- Emma Proietti Piorgo,
- Luca Colantoni,
- Giorgio Tasca,
- Mauro Monforte,
- Stefania Zampatti,
- Guido Primiano,
- Cristina Sancricca,
- Sara Bortolani,
- Eleonora Torchia,
- Beatrice Ravera,
- Francesca Torri,
- Giulio Gadaleta,
- Barbara Risi,
- Filomena Caria,
- Francesca Gerardi,
- Elena Carraro,
- Valeria Gioiosa,
- Matteo Garibaldi,
- Laura Tufano,
- Erica Frezza,
- Roberto Massa,
- Carlo Caltagirone,
- Elena Maria Pennisi,
- Antonio Petrucci,
- Marika Pane,
- Annalia Frongia,
- Francesca Gragnani,
- Marianna Scutifero,
- Paola Mandich,
- Marina Grandis,
- Maria Antonietta Maioli,
- Carlo Casali,
- Elisabetta Manfroi,
- Luisa Politano,
- Luigia Passamano,
- Roberta Petillo,
- Carmelo Rodolico,
- Alessia Pugliese,
- Stefano Carlo Previtali,
- Valeria Sansone,
- Liliana Vercelli,
- Tiziana Enrica Mongini,
- Giulia Ricci,
- Gabriele Siciliano,
- Massimiliano Filosto,
- Enzo Ricci,
- Raffaella Cascella,
- Emiliano Giardina,
- FSHD Italian Clinical Group
Affiliations
- Claudia Strafella
- Genomic Medicine Laboratory UILDM, IRCCS Santa Lucia Foundation
- Domenica Megalizzi
- Genomic Medicine Laboratory UILDM, IRCCS Santa Lucia Foundation
- Giulia Trastulli
- Genomic Medicine Laboratory UILDM, IRCCS Santa Lucia Foundation
- Emma Proietti Piorgo
- Genomic Medicine Laboratory UILDM, IRCCS Santa Lucia Foundation
- Luca Colantoni
- Genomic Medicine Laboratory UILDM, IRCCS Santa Lucia Foundation
- Giorgio Tasca
- John Walton Muscular Dystrophy Research Centre, Newcastle University and Newcastle Hospitals NHS Foundation Trusts
- Mauro Monforte
- Unità Operativa Complessa di Neurologia, Fondazione Policlinico Universitario A. Gemelli IRCCS
- Stefania Zampatti
- Genomic Medicine Laboratory UILDM, IRCCS Santa Lucia Foundation
- Guido Primiano
- Neurofisiopathology Unit, Fondazione Policlinico Universitario Agostino Gemelli IRCCS
- Cristina Sancricca
- Neurofisiopathology Unit, Fondazione Policlinico Universitario Agostino Gemelli IRCCS
- Sara Bortolani
- Unità Operativa Complessa di Neurologia, Fondazione Policlinico Universitario A. Gemelli IRCCS
- Eleonora Torchia
- Unità Operativa Complessa di Neurologia, Fondazione Policlinico Universitario A. Gemelli IRCCS
- Beatrice Ravera
- Unità Operativa Complessa di Neurologia, Fondazione Policlinico Universitario A. Gemelli IRCCS
- Francesca Torri
- Department of Clinical and Experimental Medicine, University of Pisa
- Giulio Gadaleta
- Presidio Molinette e OIRM (SS Malattie Neuromuscolari e SC Neuropsichiatria Infantile), AOU Città della Salute e della Scienza di Torino
- Barbara Risi
- NeMO-Brescia Clinical Center for Neuromuscular Diseases
- Filomena Caria
- NeMO-Brescia Clinical Center for Neuromuscular Diseases
- Francesca Gerardi
- The NEMO Center in Milan, Neurorehabilitation Unit,, University of Milan, ASST Niguarda Hospital
- Elena Carraro
- The NEMO Center in Milan, Neurorehabilitation Unit,, University of Milan, ASST Niguarda Hospital
- Valeria Gioiosa
- Department of Medico-Surgical Sciences and Biotechnologies, University of Rome Sapienza
- Matteo Garibaldi
- Neuromuscular and Rare Disease Centre, Sant’Andrea Hospital
- Laura Tufano
- Neuromuscular and Rare Disease Centre, Sant’Andrea Hospital
- Erica Frezza
- Department of Systems Medicine, Neurology Unit, Tor Vergata University of Rome
- Roberto Massa
- Department of Systems Medicine, Neurology Unit, Tor Vergata University of Rome
- Carlo Caltagirone
- Department of Clinical and Behavioral Neurology, IRCCS Fondazione Santa Lucia
- Elena Maria Pennisi
- UOC of Neurology, San Filippo Neri Hospital
- Antonio Petrucci
- Department of Neurology and Neurophysiopathology, Azienda Ospedaliera San Camillo Forlanini
- Marika Pane
- Pediatric Neurology, Università Cattolica del Sacro Cuore
- Annalia Frongia
- Centro Clinico Nemo, Fondazione Agostino Gemelli IRCCS
- Francesca Gragnani
- Neurology and Neurophysiopathology Unit, Sandro Pertini Hospital
- Marianna Scutifero
- Cardiomyology and Medical Genetics, University of Campania Luigi Vanvitelli
- Paola Mandich
- IRCCS Ospedale Policlinico San Martino - UOC Genetica Medica
- Marina Grandis
- IRCCS Ospedale Policlinico San Martino - UOC Genetica Medica
- Maria Antonietta Maioli
- Centro Sclerosi Multipla, ASSL
- Carlo Casali
- Department of Medico-Surgical Sciences and Biotechnologies, University of Rome Sapienza
- Elisabetta Manfroi
- Department of Neuroscience- Neurogenetics, Santa Maria Hospital, Viale Tristano Di Joannuccio
- Luisa Politano
- Cardiomyology and Medical Genetics, University of Campania Luigi Vanvitelli
- Luigia Passamano
- Cardiomyology and Medical Genetics, University of Campania Luigi Vanvitelli
- Roberta Petillo
- Medical and Laboratory Genetics Unit, A.O.R.N. ‘Antonio Cardarelli’
- Carmelo Rodolico
- Department of Clinical and Experimental Medicine, University of Messina
- Alessia Pugliese
- Department of Clinical and Experimental Medicine, University of Messina
- Stefano Carlo Previtali
- Institute of Experimental Neurology (INSPE), Division of Neuroscience, IRCCS San Raffaele Scientific Institute
- Valeria Sansone
- The NEMO Center in Milan, Neurorehabilitation Unit,, University of Milan, ASST Niguarda Hospital
- Liliana Vercelli
- Presidio Molinette e OIRM (SS Malattie Neuromuscolari e SC Neuropsichiatria Infantile), AOU Città della Salute e della Scienza di Torino
- Tiziana Enrica Mongini
- Presidio Molinette e OIRM (SS Malattie Neuromuscolari e SC Neuropsichiatria Infantile), AOU Città della Salute e della Scienza di Torino
- Giulia Ricci
- Department of Clinical and Experimental Medicine, University of Pisa
- Gabriele Siciliano
- Department of Clinical and Experimental Medicine, University of Pisa
- Massimiliano Filosto
- NeMO-Brescia Clinical Center for Neuromuscular Diseases
- Enzo Ricci
- Unità Operativa Complessa di Neurologia, Fondazione Policlinico Universitario A. Gemelli IRCCS
- Raffaella Cascella
- Genomic Medicine Laboratory UILDM, IRCCS Santa Lucia Foundation
- Emiliano Giardina
- Genomic Medicine Laboratory UILDM, IRCCS Santa Lucia Foundation
- FSHD Italian Clinical Group
- Italian Clinical Group for the Study of FSHD
- DOI
- https://doi.org/10.1186/s13148-024-01747-2
- Journal volume & issue
-
Vol. 16,
no. 1
pp. 1 – 13
Abstract
Abstract Background Facioscapulohumeral dystrophy (FSHD) is a myopathy characterized by the loss of repressive epigenetic features affecting the D4Z4 locus (4q35). The assessment of DNA methylation at two regions (DUX4-PAS and DR1) of D4Z4 locus proved to be an effective method to detect epigenetic signatures compatible with FSHD. The present study aims at validating the employment of this method into clinical practice and improving the protocol by refining the classification thresholds of 4qA/4qA patients. To this purpose, 218 subjects with clinical suspicion of FSHD collected in 2022–2023 were analyzed. Each participant underwent in parallel the traditional FSHD molecular testing (D4Z4 sizing) and the proposed methylation assay. The results provided by both analyses were compared to evaluate the concordance and calculate the performance metrics of the methylation test. Results Among the 218 subjects, the 4q variant type distribution was 54% 4qA/4qA, 43% 4qA/4qB and 3% 4qB/4qB. The methylation analysis was performed only on carriers of at least one 4qA allele. After refining the classification threshold, the test reached the following performance metrics: sensitivity = 0.90, specificity = 1.00 and accuracy = 0.93. These results confirmed the effectiveness of the methylation assay in identifying patients with genetic signature compatible with FSHD1 and FSHD2 based on their DUX4-PAS and DR1 profile, respectively. The methylation data were also evaluated with respect to the clinical information. Conclusions The study confirmed the ability of the method to accurately identify methylation profiles compatible with FSHD genetic signatures considering the 4q genotype. Moreover, the test allows the detection of hypomethylated profiles in asymptomatic patients, suggesting its potential application in identifying preclinical conditions in patients with positive family history and FSHD genetic signatures. Furthermore, the present work emphasizes the importance of interpreting methylation profiles considering the patients’ clinical data.
Keywords