Ciliopathies in Complex Congenital Heart Disease: Molecular Genetics, Embryologic Mechanisms and Clinical Implications Maria Felicia Gagliardi, Emanuele Micaglio, Angelo Micheletti, Sara Benedetti, Andrea Giordano, Baldassare Maria Alonzo, Giulia Guglielmi, Diana Gabriela Negura, Alessandro Giamberti, Massimo Chessa Genes, 2026 Background/Objectives: Congenital heart malformations (CHDs) are not rare diseases, and, in many cases, their pathogenic mechanisms are well established. Several conotruncal defects are associated with genetic syndromes such as DiGeorge syndrome and RASopathies, reflecting shared developmental pathways affecting cardiac outflow tract formation. However, even common CHDs may occur within complex syndromic contexts, making early diagnosis essential for optimal management. This review aims to provide a unifying framework linking ciliary dysfunction to CHD phenotypes. Methods: We performed an integrative narrative review of genetic, experimental, and developmental studies focusing on the role of primary and motile cilia in cardiac morphogenesis. Particular attention was given to signaling pathways regulated by cilia and their contribution to disease phenotypes. Results: Emerging evidence indicates that primary and motile cilia act as central regulators of cardiac development, integrating morphogen gradients and mechanical cues into transcriptional programs. Dysfunctions in ciliary structure or signaling are increasingly recognized as important contributors to selected complex CHD phenotypes, particularly in syndromic forms and laterality-associated defects. This cilia-centered model may help explain part of the phenotypic heterogeneity observed in CHD and highlights shared mechanisms across distinct clinical entities. Conclusions: Understanding cilia-dependent mechanisms provides a unifying conceptual framework linking genetic defects to disrupted morphogenesis. This perspective may refine disease interpretation and support future development of precision diagnostics and pathway-informed therapeutic strategies in CHD.
Tetralogy of Fallot: Genetic, Epigenetic and Clinical Insights into a Multifactorial Congenital Heart Disease Maria Felicia Gagliardi, Emanuele Micaglio, Angelo Micheletti, Sara Benedetti, Diana Gabriela Negura, Francesca Bevilacqua, Giulia Guglielmi, Giulia Pasqualin, Alessandro Giamberti, Massimo Chessa Genes, 2026 Tetralogy of Fallot (TOF) is the most common cyanotic congenital heart disease, classically characterized by right ventricular outflow tract obstruction, ventricular septal defect, overriding aorta, and right ventricular hypertrophy. Recent advances in molecular and genomic research indicate that TOF is part of a phenotypic continuum encompassing Trilogy, Tetralogy, and Pentalogy of Fallot, in which the variability of anatomical presentation reflects shared genetic and epigenetic mechanisms with highly variable penetrance and expressivity. Variants in NOTCH1, FLT4, KDR, GATA6, and TBX1 highlight key pathways in conotruncal development and endothelial–mesenchymal transition, yet these well-known genes explain only a fraction of the genetic landscape. Emerging studies have identified additional candidate genes and networks involved in cardiac morphogenesis, including transcriptional regulators, signaling mediators, chromatin-remodeling factors, and splicing-associated genes such as PUF60 and DVL3. Epigenetic mechanisms, including DNA methylation, histone modifications, and non-coding RNA expression, further modulate phenotypic expressivity and contribute to variability along the Trilogy–Tetralogy–Pentalogy spectrum. This review integrates current genomic and clinical evidence to provide a comprehensive overview of the molecular architecture of Fallot-type conotruncal malformations, emphasizing the interplay between genetic and epigenetic mechanisms, genotype–phenotype correlations, and implications for diagnosis, risk stratification, counseling, and personalized management in the era of precision cardiology.
Dropped Head Syndrome Unmasking Myotonic Dystrophy Type 1 in a Patient with Parkinson's Disease: A Case Report and a Case-Based Review Michele Giovanni Croce, Francesca Valentino, Emanuele Micaglio, Sara Benedetti, Matteo Paoletti, Giuseppe Cosentino, Sabrina Ravaglia Movement Disorders Clinical Practice, 2026 Ethical Compliance Statement: Informed patient consent was regularly obtained for video usage. Both oral and written consent was obtained. The approval of an institutional review board was not required. We confirm that we have read the Journal's position on issues involved in ethical publication and affirm that this work is consistent with those guidelines. Funding Sources and Conflicts of Interest: No specific funding was received for this work. The authors declare that there are no conflicts of interest relevant to this work. Financial Disclosures for the Previous 12 Months: The authors declare that there are no additional disclosures to report. The data that support the findings of this study are available from the corresponding author upon reasonable request. TABLE S1. Clinical, neurophysiological, neuroimaging and therapeutic features of reported cases of myotonic dystrophy and concurrent parkinsonian syndrome (ref. 1–10) and of reported cases of myotonic dystrophy presenting with dropped head syndrome (ref 11,12). AAO, age at onset; DM, myotonic dystrophy; DHS, dropped head syndrome; EMG, electromyography; PET, positron emission tomography; SPECT, single-photon emission computed tomography; DAT, dopamine transporter; MSA-P, multiple system atrophy, parkinsonian variant. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
Prevalence of rare missense TTN variants in a cohort of patients with cardiomyopathy Irene Bottillo, Maria Pia Ciccone, Monia Magliozzi, Kalliopi Pilichou, Giorgia Girotto, Francesca Girolami, Massimiliano Cecconi, Valeria D'Argenio, Valeria Novelli, Alessandra Coiana, Daniela Formicola, Emanuele Micaglio, Giada Tortora, Francesca Gualandi, Simona Petrucci, Marco Castori, Nicoletta Resta, Anna Rita Vestri, Maria Iascone, Paola Grammatico Journal of Molecular and Cellular Cardiology, 2025
When Paying Attention Pays Back: Missense Mutation c.1006G>A p. (Val336Ile) in PRKAG2 Gene Causing Left Ventricular Hypertrophy and Conduction Abnormalities in a Caucasian Patient: Case Report and Literature Review Emanuele Micaglio, Lara Tondi, Sara Benedetti, Maria Alessandra Schiavo, Antonia Camporeale, Giandomenico Disabato, Andrea Attanasio, Gianluigi Guida, Gianpaolo Carrafiello, Massimo Piepoli, Pietro Spagnolo, Carlo Pappone, Massimo Lombardi International Journal of Molecular Sciences, 2024 PRKAG2 cardiomyopathy is a rare genetic disorder that manifests early in life with an autosomal dominant inheritance pattern. It harbors left ventricular hypertrophy (LVH), ventricular pre-excitation and progressively worsening conduction system defects. Its estimated prevalence among patients with LVH ranges from 0.23 to about 1%, but it is likely an underdiagnosed condition. We report the association of the PRKAG2 missense variant c.1006G>A p. (Val336Ile) with LVH, conduction abnormalities (short PR interval and incomplete right bundle branch bock) and early-onset arterial hypertension (AH) in a 44-year-old Caucasian patient. While cardiac magnetic resonance (CMR) showed a mild hypertrophic phenotype with maximal wall thickness of 17 mm in absence of tissue alterations, the electric phenotype was relevant including brady–tachy syndrome and recurrent syncope. The same variant has been detected in the patient’s sister and daughter, with LVH + early-onset AH and electrocardiographic (ECG) alterations + lipothymic episodes, respectively. Paying close attention to the coexistence of LVH and ECG alterations in the proband has been helpful in directing genetic tests to exclude primary cardiomyopathy. Hence, identifying the genetic basis in the patient allowed for familial screening as well as a proper follow-up and therapeutic management of the affected members. A review of the PRKAG2 cardiomyopathy literature is provided alongside the case report.
Towards genomic-Newborn Screening: Technical feasibility of Exome Sequencing starting from dried blood spots Alessia Mauri, Clarissa Berardo, Davide Biganzoli, Andrea Meta, Sara Benedetti, Federica Rey, Letizia Messa, Gian Vincenzo Zuccotti, Stephana Carelli, Luisella Alberti, Cristina Cereda Molecular Genetics and Metabolism Reports, 2024 Each year thousands of babies are born with rare genetic disorders not identified by current NBS panels, due to programs which are not yet optimal. Next-generation sequencing technologies have the potential to overcome many NBS drawbacks and provide large amounts of molecular data, broadening the number of diseases investigated. Here, we design and set up an NGS-based approach to evaluate the feasibility of NGS from dried blood spot starting from 34 DBSs. After assessing gDNA yield and integrity, libraries were performed using three target enrichment approaches, sequenced on NS500 platform, and analyzed on commercial platform. Specifically, we focus on virtual gene panels related to highly actionable neonatal/pediatric disorders. WES show that amount and quality of DBS-extracted gDNA are suitable for high-throughput sequencing. We obtain 500-1500 ng for each specimen, 1.7-1.8 260/280 wavelength, and DIN of 7 resulting DNA integrity, on par with traditional venous blood collection. A high read depth with 94.3% coverage uniformity is achieved for all samples. Data results on mean coverage are comparable among the different workflows tested and demonstrate that DBS from newborn collected at birth is a suitable material for the developing of gNBS programs.
Multimodal Detection and Targeting of Biopsy-Proven Myocardial Inflammation in Genetic Cardiomyopathies: A Pilot Report Giovanni Peretto, Giacomo De Luca, Andrea Villatore, Chiara Di Resta, Simone Sala, Anna Palmisano, Davide Vignale, Corrado Campochiaro, Davide Lazzeroni, Monica De Gaspari, Stefania Rizzo, Elena Busnardo, Paola Ferro, Luigi Gianolli, Cristina Basso, Lorenzo Dagna, Antonio Esposito, Sara Benedetti, Paolo Della Bella Jacc Basic to Translational Science, 2023 The authors present a clinical report focused on the overlap between myocarditis and genetic cardiomyopathies of the dilated and arrhythmogenic spectrum. Our cohort was composed of 25 patients undergoing extensive baseline characterization and prospective reassessment by a dedicated multidisciplinary disease unit during a median follow-up of 69 months. We showed that the use of multimodal imaging allowed both discrimination of specific genotypes and identification of myocardial inflammation proven using endomyocardial biopsy. In addition, we showed that the use of immunomodulatory therapy was beneficial for most patients.
Functional Characterisation of the Rare SCN5A p.E1225K Variant, Segregating in a Brugada Syndrome Familial Case, in Human Cardiomyocytes from Pluripotent Stem Cells Nicolò Salvarani, Giovanni Peretto, Crasto Silvia, Andrea Villatore, Cecilia Thairi, Anna Santoni, Camilla Galli, Paola Carrera, Simone Sala, Sara Benedetti, Elisa Di Pasquale, Chiara Di Resta International Journal of Molecular Sciences, 2023 Brugada syndrome (BrS) is an inherited autosomal dominant cardiac channelopathy. Pathogenic rare mutations in the SCN5A gene, encoding the alpha-subunit of the voltage-dependent cardiac Na+ channel protein (Nav1.5), are identified in 20% of BrS patients, affecting the correct function of the channel. To date, even though hundreds of SCN5A variants have been associated with BrS, the underlying pathogenic mechanisms are still unclear in most cases. Therefore, the functional characterization of the SCN5A BrS rare variants still represents a major hurdle and is fundamental to confirming their pathogenic effect. Human cardiomyocytes (CMs) differentiated from pluripotent stem cells (PSCs) have been extensively demonstrated to be reliable platforms for investigating cardiac diseases, being able to recapitulate specific traits of disease, including arrhythmic events and conduction abnormalities. Based on this, in this study, we performed a functional analysis of the BrS familial rare variant NM_198056.2:c.3673G>A (NP_932173.1:p.Glu1225Lys), which has been never functionally characterized before in a cardiac-relevant context, as the human cardiomyocyte. Using a specific lentiviral vector encoding a GFP-tagged SCN5A gene carrying the specific c.3673G>A variant and CMs differentiated from control PSCs (PSC-CMs), we demonstrated an impairment of the mutated Nav1.5, thus suggesting the pathogenicity of the rare BrS detected variant. More broadly, our work supports the application of PSC-CMs for the assessment of the pathogenicity of gene variants, the identification of which is increasing exponentially due to the advances in next-generation sequencing methods and their massive use in genetic testing.
Brugada syndrome genetics is associated with phenotype severity Giuseppe Ciconte, Michelle M Monasky, Vincenzo Santinelli, Emanuele Micaglio, Gabriele Vicedomini, Luigi Anastasia, Gabriele Negro, Valeria Borrelli, Luigi Giannelli, Francesca Santini, Carlo de Innocentiis, Roberto Rondine, Emanuela T Locati, Andrea Bernardini, Beniamino C Mazza, Valerio Mecarocci, Žarko Ćalović, Andrea Ghiroldi, Sara D’Imperio, Sara Benedetti, Chiara Di Resta, Ilaria Rivolta, Giorgio Casari, Enrico Petretto, Carlo Pappone European Heart Journal, 2021
Immunosuppressive therapy in childhood-onset arrhythmogenic inflammatory cardiomyopathy Giovanni Peretto, Federica Barzaghi, Maria Pia Cicalese, Chiara Di Resta, Massimo Slavich, Sara Benedetti, Sara Giangiobbe, Stefania Rizzo, Anna Palmisano, Antonio Esposito, Francesco De Cobelli, Simone Gulletta, Cristina Basso, Giorgio Casari, Alessandro Aiuti, Paolo Della Bella, Simone Sala PACE Pacing and Clinical Electrophysiology, 2021
Novel scn5a p.V1429m variant segregation in a family with brugada syndrome Michelle M. Monasky, Emanuele Micaglio, Giuseppe Ciconte, Valeria Borrelli, Luigi Giannelli, Gabriele Vicedomini, Andrea Ghiroldi, Luigi Anastasia, Emanuela T. Locati, Sara Benedetti, Chiara Di Resta, Giorgio Casari, Carlo Pappone International Journal of Molecular Sciences, 2020
Novel scn5a p.W697x nonsense mutation segregation in a family with brugada syndrome Emanuele Micaglio, Michelle Monasky, Nicoletta Resta, Rosanna Bagnulo, Giuseppe Ciconte, Luigi Giannelli, Emanuela Locati, Gabriele Vicedomini, Valeria Borrelli, Andrea Ghiroldi, Luigi Anastasia, Sara Benedetti, Chiara Di Resta, Maurizio Ferrari, Carlo Pappone International Journal of Molecular Sciences, 2019
Comparable clinical characteristics in Brugada syndrome patients harboring SCN5A or novel SCN10A variants Michelle M Monasky, Emanuele Micaglio, Gabriele Vicedomini, Emanuela T Locati, Giuseppe Ciconte, Luigi Giannelli, Federica Giordano, Simonetta Crisà, Mattia Vecchi, Valeria Borrelli, Andrea Ghiroldi, Sara D'Imperio, Chiara Di Resta, Sara Benedetti, Maurizio Ferrari, Vincenzo Santinelli, Luigi Anastasia, Carlo Pappone Europace, 2019
Cardiac and neuromuscular features of patients with LMNA-related cardiomyopathy Giovanni Peretto, Chiara Di Resta, Jacopo Perversi, Cinzia Forleo, Lorenzo Maggi, Luisa Politano, Andrea Barison, Stefano C. Previtali, Nicola Carboni, Francesca Brun, Elena Pegoraro, Adele D'Amico, Carmelo Rodolico, Francesca Magri, Rosa C. Manzi, Alberto Palladino, Franco Isola, Lorenzo Gigli, Tiziana E. Mongini, Claudio Semplicini, Chiara Calore, Giulia Ricci, Giacomo P. Comi, Lucia Ruggiero, Enrico Bertini, Paolo Bonomo, Gerardo Nigro, Nicoletta Resta, Michele Emdin, Stefano Favale, Gabriele Siciliano, Lucio Santoro, Gianfranco Sinagra, Giuseppe Limongelli, Alessandro Ambrosi, Maurizio Ferrari, Pier G. Golzio, Paolo Della Bella, Sara Benedetti, Simone Sala, and Annals of Internal Medicine, 2019
SCN5A nonsense mutation and NF1 frameshift mutation in a family with brugada syndrome and neurofibromatosis Emanuele Micaglio, Michelle M. Monasky, Giuseppe Ciconte, Gabriele Vicedomini, Manuel Conti, Valerio Mecarocci, Luigi Giannelli, Federica Giordano, Alberto Pollina, Massimo Saviano, Simonetta Crisà, Valeria Borrelli, Andrea Ghiroldi, Sara D’Imperio, Chiara Di Resta, Sara Benedetti, Maurizio Ferrari, Vincenzo Santinelli, Luigi Anastasia, Carlo Pappone Frontiers in Genetics, 2019
Novel SCN5A frameshift mutation in Brugada syndrome associated with complex arrhythmic phenotype Emanuele Micaglio, Michelle M. Monasky, Giuseppe Ciconte, Gabriele Vicedomini, Manuel Conti, Valerio Mecarocci, Luigi Giannelli, Federica Giordano, Alberto Pollina, Massimo Saviano, Paolo R. Pozzi, Chiara Di Resta, Sara Benedetti, Maurizio Ferrari, Vincenzo Santinelli, Carlo Pappone Frontiers in Genetics, 2019
Emerging perspectives on laminopathies Luisa Politano, Giovanna Lattanzi, Sara Benedetti, Maria Rosaria D'Apice, Lorenzo Maggi, Nicola Carboni, Emanuela Scarano Cell Health and Cytoskeleton, 2016
LMNA-associated myopathies: The Italian experience in a large cohort of patients L. Maggi, A. D'Amico, A. Pini, S. Sivo, M. Pane, G. Ricci, L. Vercelli, P. D'Ambrosio, L. Travaglini, S. Sala, G. Brenna, D. Kapetis, M. Scarlato, E. Pegoraro, M. Ferrari, A. Toscano, S. Benedetti, P. Bernasconi, L. Colleoni, G. Lattanzi, E. Bertini, E. Mercuri, G. Siciliano, C. Rodolico, T. Mongini, L. Politano, S. C. Previtali, N. Carboni, R. Mantegazza, L. Morandi Neurology, 2014
Analyzing histopathological features of rare Charcot-Marie-Tooth neuropathies to unravel their pathogenesis Sara Benedetti, Stefano Carlo Previtali, Silvia Coviello, Marina Scarlato, Federica Cerri, Emanuela Di Pierri, Lara Piantoni, Ivana Spiga, Raffaella Fazio, Nilo Riva, Maria Grazia Natali Sora, Patrizia Dacci, Maria Chiara Malaguti, Elisabetta Munerati, Luigi Maria Edoardo Grimaldi, Maria Giovanna Marrosu, Maurizio De Pellegrin, Maurizio Ferrari, Giancarlo Comi, Angelo Quattrini, Alessandra Bolino Archives of Neurology, 2010
Phenotypic clustering of lamin A/C mutations in neuromuscular patients S. Benedetti, I. Menditto, M. Degano, C. Rodolico, L. Merlini, A. D’Amico, L. Palmucci, A. Berardinelli, E. Pegoraro, C. P. Trevisan, L. Morandi, I. Moroni, G. Galluzzi, E. Bertini, A. Toscano, M. Olivè, G. Bonne, F. Mari, R. Caldara, R. Fazio, I. Mammì, P. Carrera, D. Toniolo, G. Comi, A. Quattrini, M. Ferrari, S. C. Previtali Neurology, 2007
Gene therapy of experimental brain tumors using neural progenitor cells Sara Benedetti, Barbara Pirola, Bianca Pollo, Lorenzo Magrassi, Maria Grazia Bruzzone, Dorotea Rigamonti, Rossella Galli, Silvia Selleri, Francesco Di Meco, Claudio De Fraja, Angelo Vescovi, Elena Cattaneo, Gaetano Finocchiaro Nature Medicine, 2000
Eradication of rat malignant gliomas by retroviral-mediated, in vivo delivery of the interleukin 4 gene Cancer Research, 1999
IL-4 gene transfer for the treatment of experimental gliomas S. Benedetti, F. Di Meco, N. Cirenei, M. G. Bruzzone, B. Pollo, N. Florio, L. Caposio, M. P. Colombo, E. Cattaneo, Gaetano Finocchiaro Advances in Experimental Medicine and Biology, 1998
The "bystander effect": Association of U-87 cell death with ganciclovir-mediated apoptosis of nearby cells and lack of effect in athymic mice Human Gene Therapy, 1995
