Biochemistry, Genetics and Molecular Biology, Biochemistry (medical)
46
Scopus Publications
Scopus Publications
Over ten years of newborn screening for LSDs in Tuscany (Italy): Epidemiology, novel variants, and the pseudodeficiency burden Sabrina Malvagia, Marta Daniotti, Rodolfo Tonin, Lorenzo Ferri, Daniela Ombrone, Giulia Forni, Emanuela Scolamiero, Silvia Funghini, Marina Rinaldi, Silvia Falliano, Antonella Paoli, Massimo Mura, Francesca Raspini, Sara Poggiali, Giusi Scaturro, Michele Sacchini, Maria Alice Donati, Elena Procopio, Renzo Guerrini, Amelia Morrone, Giancarlo la Marca Molecular Genetics and Metabolism, 2026
Newborn Screening for Metachromatic Leukodystrophy in Tuscany: The Paradigm of a Successful Preventive Medicine Program Sabrina Malvagia, Alessandra Bettiol, Margherita Porcaro, Massimo Mura, Silvia Funghini, Daniela Ombrone, Giulia Forni, Emanuela Scolamiero, Filippo Coppi, Roberta Damiano, Cristina Cereda, Simonetta Simonetti, Annalisa Lonetti, Marta Daniotti, Anna Caciotti, Amelia Morrone, Valeria Calbi, Francesca Fumagalli, Alessandro Aiuti, Elena Procopio, Renzo Guerrini, Giancarlo la Marca International Journal of Neonatal Screening, 2025 Metachromatic leukodystrophy (MLD) is a rare inherited disorder of lysosomal storage, caused by a deficiency in the arylsulfatase A (ARSA) enzyme, leading to toxic accumulation of sulfatides, which progressively impair motor and cognitive function. MLD is a candidate for inclusion in newborn screening (NBS) programs, due to the narrow pre-symptomatic window for effective therapeutic intervention. We set up a prospective pilot NBS program for MLD in Tuscany, based on a two-step approach. The first-tier test quantified four sulfatides; if levels exceeded the cut-off, we performed the second-tier test by measuring ARSA activity on the same neonatal dried blood spot (DBS). We performed the first-tier test on 42,262 newborns over two years and the second-tier test on residual neonatal DBS from 90 of them (0.21%). We recalled 10 newborns (0.02%) for an additional DBS, due to insufficient residual material for a second-tier test (n = 4) or to low ARSA activity (n = 6). We found normal ARSA activity in all new DBS and identified no new cases of MLD. Retrospective analysis of eight neonatal and fifteen non-neonatal DBS from patients with genetically confirmed MLD showed that the algorithm accurately identified MLD patients. This diagnostic algorithm proved feasible and accurate for early detection of MLD in prospective NBS.
Multi-year enzyme expression in patients with mucopolysaccharidosis type VI after liver-directed gene therapy Alessandro Rossi, Roberta Romano, Simona Fecarotta, Margherita Dell’Anno, Valentina Pecorella, Roberta Passeggio, Stefano Zancan, Giancarlo Parenti, Francesca Santamaria, Francesco Borgia, Federica Deodato, Silvia Funghini, Charles A. Rupar, Chitra Prasad, Mar O’Callaghan, John J. Mitchell, Maria Grazia Valsecchi, Giancarlo la Marca, Stefania Galimberti, Alberto Auricchio, Nicola Brunetti-Pierri Med, 2025 Background Mucopolysaccharidosis type VI (MPS VI) is due to a deficiency of the lysosomal enzyme arylsulfatase B (ARSB) that results in multi-organ accumulation of glycosaminoglycans (GAGs). Limitations of current treatments prompted the development of a liver-directed gene therapy clinical trial for MPS VI. Methods We report the long-term follow-up of patients with MPS VI who discontinued enzyme replacement therapy (ERT) and received a single intravenous infusion of high-dose (6 × 10 12 genome copies/kg) recombinant adeno-associated virus serotype 8 (AAV8) vector expressing ARSB under the control of a liver-specific promoter ( ClinicalTrials.gov : NCT03173521 ). Primary outcomes were safety and urinary GAG excretion. Secondary outcomes were endurance and respiratory function. Findings Median follow-up time was 45 months ( n = 4, three females and one male; age range: 5–10 years). No late-emergent safety events were observed. Patients showed sustained serum ARSB activity (38%–67% of mean healthy reference values), a modest increase in urinary GAG concentrations, and no relevant changes in endurance, cardiac, or pulmonary function. In one of the four patients, ERT was restarted because of elevated urinary GAGs without decreased serum ARSB activity up to about 2.5 years after gene transfer. Liver and spleen size remained within the reference ranges. Conclusions A single intravenous administration of AAV8.TBG.hARSB was safe and resulted in sustained ARSB expression and a modest increase in urinary GAGs in most patients, thus supporting liver-directed gene therapy for MPS VI. Funding This study was sponsored by the Telethon Foundation ETS, the European Union, the Isaac Foundation, and the Italian Ministry of University and Research. • Long-term data of AAV-mediated liver-directed gene therapy in patients with MPS VI • Sustained ARSB expression and modest increase in urine glycosaminoglycans • Endurance, pulmonary and cardiac function showed no changes despite ERT interruption Mucopolysaccharidosis type VI (MPS VI) is a rare lysosomal storage disease due to arylsulfatase B (ARSB) deficiency that leads to a multisystem accumulation of glycosaminoglycans (GAGs). Current treatment based on weekly infusions of enzyme replacement therapy (ERT) is unsatisfactory. In this study, after the discontinuation of ERT, MPS VI individuals received a single intravenous infusion of an AAV vector that delivered the gene encoding ARSB to the liver. AAV vector administration was safe and resulted in sustained ARSB expression for at least 3 years post-injection. Although a modest increase in urine GAGs was detected, no significant changes in endurance, liver and spleen sizes, or pulmonary and cardiac functions were observed. These findings support the safety and efficacy of AAV-mediated liver-directed gene therapy for MPS VI. Rossi et al. presented longitudinal data on mucopolysaccharidosis type VI individuals who discontinued enzyme replacement therapy and received AAV-mediated liver-directed gene therapy. Sustained serum ARSB expression and a modest increase in urine glycosaminoglycans were observed. No significant changes were detected in endurance, or pulmonary and cardiac function.
Expanded Newborn Screening in Italy Using Tandem Mass Spectrometry: Two Years of National Experience Margherita Ruoppolo, Sabrina Malvagia, Sara Boenzi, Carla Carducci, Carlo Dionisi-Vici, Francesca Teofoli, Alberto Burlina, Antonio Angeloni, Tommaso Aronica, Andrea Bordugo, Ines Bucci, Marta Camilot, Maria Teresa Carbone, Roberta Cardinali, Claudia Carducci, Michela Cassanello, Cinzia Castana, Chiara Cazzorla, Renzo Ciatti, Simona Ferrari, Giulia Frisso, Silvia Funghini, Francesca Furlan, Serena Gasperini, Vincenza Gragnaniello, Chiara Guzzetti, Giancarlo La Marca, Luisa La Spina, Tania Lorè, Concetta Meli, MariaAnna Messina, Amelia Morrone, Francesca Nardecchia, Rita Ortolano, Giancarlo Parenti, Enza Pavanello, Damiana Pieragostino, Sara Pillai, Francesco Porta, Francesca Righetti, Claudia Rossi, Valentina Rovelli, Alessandro Salina, Laura Santoro, Pina Sauro, Maria Cristina Schiaffino, Simonetta Simonetti, Monica Vincenzi, Elisabetta Tarsi, Anna Paola Uccheddu International Journal of Neonatal Screening, 2022 Newborn screening (NBS) for inborn errors of metabolism is one of the most advanced tools for secondary prevention in medicine, as it allows early diagnosis and prompt treatment initiation. The expanded newborn screening was introduced in Italy between 2016 and 2017 (Law 167/2016; DM 13 October 2016; DPCM 12-1-2017). A total of 1,586,578 infants born in Italy were screened between January 2017 and December 2020. For this survey, we collected data from 15 Italian screening laboratories, focusing on the metabolic disorders identified by tandem mass spectrometry (MS/MS) based analysis between January 2019 and December 2020. Aminoacidemias were the most common inborn errors in Italy, and an equal percentage was observed in detecting organic acidemias and mitochondrial fatty acids beta-oxidation defects. Second-tier tests are widely used in most laboratories to reduce false positives. For example, second-tier tests for methylmalonic acid and homocysteine considerably improved the screening of CblC without increasing unnecessary recalls. Finally, the newborn screening allowed us to identify conditions that are mainly secondary to a maternal deficiency. We describe the goals reached since the introduction of the screening in Italy by exchanging knowledge and experiences among the laboratories.
Lysosomals Silvia Funghini, Sabrina Malvagia, Giulia Polo, Giancarlo la Marca Physician S Guide to the Diagnosis Treatment and Follow Up of Inherited Metabolic Diseases Second Edition, 2022
The successful inclusion of ADA SCID in Tuscany expanded newborn screening program Sabrina Malvagia, Silvia Funghini, Maria Della Bona, Daniela Ombrone, Massimo Mura, Roberta Damiano, Silvia Ricci, Martina Cortimiglia, Chiara Azzari, Giancarlo la Marca Clinical Chemistry and Laboratory Medicine, 2021 Adenosine deaminase (ADA) deficiency is a known cause for severe combined immunodeficiency (SCID). Various ADA deficiency phenotypes have been described depending on the clinical onset and on the severity of the disease. We report a new case of ADA-deficient patient with delayed-onset identified by Tuscany newborn screening program with tandem mass spectrometry method while both TRECs and KRECs repeatedly proved to be within normal limits.
High frequency of biotinidase deficiency in Italian population identified by newborn screening Silvia Funghini, Rodolfo Tonin, Sabrina Malvagia, Anna Caciotti, Maria Alice Donati, Amelia Morrone, Giancarlo la Marca Molecular Genetics and Metabolism Reports, 2020 The biotinidase (BTD) enzyme is essential for recycling biotin, a water-soluble B-complex vitamin that is the coenzyme of four carboxylases involved in fatty acid synthesis, amino acid catabolism and gluconeogenesis. If untreated, total or partial BTD deficiencies lead to an autosomal recessive inherited organic aciduria whose clinical features, mainly presenting in the first years of life, include, seizures, skin rash, and alopecia. Based on residual BTD enzyme activity it is possible to identify partial or total biotinidase deficiency. The incidence of profound and partial biotinidase deficiency worldwide is estimated to be about 1 in 60.000. We report twelve years of experience in the newborn screening of biotinidase deficiency on 466.182 neonates. When a positive screening result occurred, a clinical evaluation was made of the patient and genetic counselling was offered to the family. Molecular analysis the BTD gene was carried out in all recalled neonates. Newborn screening lead to the identification of 75 BTD deficiencies with an incidence of about 1:6.300 births, ten times higher than the reported worldwide incidence. BTD deficiency was confirmed at a genomic level in all patients, demonstrating a high frequency of the p.(Asp444His) amino acid substitution and the complex allele p.(Ala171Thr)/p.(Asp444His) in the analyzed Italian newborns. Four new mutations (two small deletions, one stop mutation and one missense mutation) and a new combined allelic alteration were identified. Our data suggests that there is a high incidence of the biotinidase defect in the Italian population, most likely due to the high frequency of certain mutations.
A new strategy implementing mass spectrometry in the diagnosis of congenital disorders of N-glycosylation (CDG) Bruno Casetta, Sabrina Malvagia, Silvia Funghini, Diego Martinelli, Carlo Dionisi-Vici, Rita Barone, Agata Fiumara, Maria Alice Donati, Renzo Guerrini, Giancarlo la Marca Clinical Chemistry and Laboratory Medicine, 2020 Objectives Congenital disorders of N-glycosylation (CDG) are a large group of rare metabolic disorders caused by defects in the most common post-translational modification of proteins. CDGs are often difficult to diagnose as they are manifested with non-specific symptoms and signs. Analysis of serum transferrin (TRF) isoforms, as the classical procedure used to identify a CDG patient, enables to predict pathological steps in the N-linked glycosylation process. Methods We devised a new strategy based on liquid chromatography-mass spectrometry (LC-MS) for the analysis of TRF isoforms by combining a simple and fast sample preparation with a specific chromatographic cleanup/separation step followed by mass-spectrometric measurement. Single TRF isoform masses were obtained through reconstruction of multiply charged electrospray data collected by quadrupole-MS technology. Hereby, we report the first analyzed serum samples obtained from 20 CDG patients and 100 controls. Results The ratio of desialylated isoforms to total TRF was calculated for patients and controls. CDG-Type I patients showed higher amounts of bi-sialo isoform (range: 6.7–29.6%) compared to controls (<5.5%, mean percentage 3.9%). CDG-Type II pattern showed an increased peak of tri-sialo isoforms. The mean percentage of tri-sialo-TRF was 9.3% (range: 2.9–12.9%) in controls, which was lower than that obtained from two patients with COG5-CDG and MAN1B1-CDG (18.5 and 24.5%). Intraday and between-day imprecisions were less than 9 and 16%, respectively, for bi-sialo- and less than 3 and 6% for tri-sialo-TRF. Conclusions This LC-MS-based approach provides a simple, sensitive and fast analytical tool for characterizing CDG disorders in a routine clinical biochemistry while improving diagnostic accuracy and speeding clinical decision-making.
The inclusion of ADA-SCID in expanded newborn screening by tandem mass spectrometry Giancarlo la Marca, Elisa Giocaliere, Sabrina Malvagia, Silvia Funghini, Daniela Ombrone, Maria Luisa Della Bona, Clementina Canessa, Francesca Lippi, Francesca Romano, Renzo Guerrini, Massimo Resti, Chiara Azzari Journal of Pharmaceutical and Biomedical Analysis, 2014
Diagnosis of immunodeficiency caused by a purine nucleoside phosphorylase defect by using tandem mass spectrometry on dried blood spots Giancarlo la Marca, Clementina Canessa, Elisa Giocaliere, Francesca Romano, Sabrina Malvagia, Silvia Funghini, Maria Moriondo, Claudia Valleriani, Francesca Lippi, Daniela Ombrone, Maria Luisa Della Bona, Carsten Speckmann, Stephan Borte, Nicholas Brodszki, Andrew R. Gennery, Katja Weinacht, Fatih Celmeli, Julia Pagel, Maurizio de Martino, Renzo Guerrini, Helmut Wittkowski, Ines Santisteban, Pawan Bali, Aydan Ikinciogullari, Michael Hershfield, Luigi D. Notarangelo, Massimo Resti, Chiara Azzari Journal of Allergy and Clinical Immunology, 2014
Tandem mass spectrometry, but not T-cell receptor excision circle analysis, identifies newborns with late-onset adenosine deaminase deficiency Giancarlo la Marca, Clementina Canessa, Elisa Giocaliere, Francesca Romano, Marzia Duse, Sabrina Malvagia, Francesca Lippi, Silvia Funghini, Leila Bianchi, Maria Luisa Della Bona, Claudia Valleriani, Daniela Ombrone, Maria Moriondo, Fabio Villanelli, Carsten Speckmann, Stuart Adams, Bobby H. Gaspar, Michael Hershfield, Ines Santisteban, Lynette Fairbanks, Giovanni Ragusa, Massimo Resti, Maurizio de Martino, Renzo Guerrini, Chiara Azzari Journal of Allergy and Clinical Immunology, 2013
New clinical and molecular insights on Barth syndrome. Lorenzo Ferri, Maria Alice Donati, Silvia Funghini, Sabrina Malvagia, Serena Catarzi, Licia Lugli, Luca Ragni, Enrico Bertini, Frédéréc M Vaz, David N Cooper, Renzo Guerrini, Amelia Morrone Orphanet Journal of Rare Diseases, 2013
Silent increase of urinary ethylmalonic acid is an indicator of nonspecific brain dysfunction Raffaella Zannolli, Sabrina Buoni, Maria Tassini, Anna De Nicola, Gianni Betti, Claudio De Felice, Alessandra Orsi, Maria Concetta Varetti, Francesco Ferrara, Mario Messina, Cosimo Giannini, Angelika Mohn, Francesco Chiarelli, Marco Liberati, Mirella Strambi, Silvia Funghini, Antonio Vivi, Ron A. Wevers, Joseph Hayek NMR in Biomedicine, 2010
