63 results on '"Valerio Conti"'
Search Results
2. Morphometric network-based abnormalities correlate with psychiatric comorbidities and gene expression in PCDH19-related developmental and epileptic encephalopathy
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Matteo Lenge, Simona Balestrini, Antonio Napolitano, Davide Mei, Valerio Conti, Giulia Baldassarri, Marina Trivisano, Simona Pellacani, Letizia Macconi, Daniela Longo, Maria Camilla Rossi Espagnet, Simona Cappelletti, PCDH19 Clinical Study Group, Ludovico D’Incerti, Carmen Barba, Nicola Specchio, and Renzo Guerrini
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Neurosciences. Biological psychiatry. Neuropsychiatry ,RC321-571 - Abstract
Abstract Protocadherin-19 (PCDH19) developmental and epileptic encephalopathy causes an early-onset epilepsy syndrome with limbic seizures, typically occurring in clusters and variably associated with intellectual disability and a range of psychiatric disorders including hyperactive, obsessive-compulsive and autistic features. Previous quantitative neuroimaging studies revealed abnormal cortical areas in the limbic formation (parahippocampal and fusiform gyri) and underlying white-matter fibers. In this study, we adopted morphometric, network-based and multivariate statistical methods to examine the cortex and substructure of the hippocampus and amygdala in a cohort of 20 PCDH19-mutated patients and evaluated the relation between structural patterns and clinical variables at individual level. We also correlated morphometric alterations with known patterns of PCDH19 expression levels. We found patients to exhibit high-significant reductions of cortical surface area at a whole-brain level (left/right p value = 0.045/0.084), and particularly in the regions of the limbic network (left/right parahippocampal gyri p value = 0.230/0.016; left/right entorhinal gyri p value = 0.002/0.327), and bilateral atrophy of several subunits of the amygdala and hippocampus, particularly in the CA regions (head of the left CA3 p value = 0.002; body of the right CA3 p value = 0.004), and differences in the shape of hippocampal structures. More severe psychiatric comorbidities correlated with more significant altered patterns, with the entorhinal gyrus (p value = 0.013) and body of hippocampus (p value = 0.048) being more severely affected. Morphometric alterations correlated significantly with the known expression patterns of PCDH19 (r value = -0.26, p spin = 0.092). PCDH19 encephalopathy represents a model of genetically determined neural network based neuropsychiatric disease in which quantitative MRI-based findings correlate with the severity of clinical manifestations and had have a potential predictive value if analyzed early.
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- 2024
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3. Structural and non-coding variants increase the diagnostic yield of clinical whole genome sequencing for rare diseases
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Alistair T. Pagnamenta, Carme Camps, Edoardo Giacopuzzi, John M. Taylor, Mona Hashim, Eduardo Calpena, Pamela J. Kaisaki, Akiko Hashimoto, Jing Yu, Edward Sanders, Ron Schwessinger, Jim R. Hughes, Gerton Lunter, Helene Dreau, Matteo Ferla, Lukas Lange, Yesim Kesim, Vassilis Ragoussis, Dimitrios V. Vavoulis, Holger Allroggen, Olaf Ansorge, Christian Babbs, Siddharth Banka, Benito Baños-Piñero, David Beeson, Tal Ben-Ami, David L. Bennett, Celeste Bento, Edward Blair, Charlotte Brasch-Andersen, Katherine R. Bull, Holger Cario, Deirdre Cilliers, Valerio Conti, E. Graham Davies, Fatima Dhalla, Beatriz Diez Dacal, Yin Dong, James E. Dunford, Renzo Guerrini, Adrian L. Harris, Jane Hartley, Georg Hollander, Kassim Javaid, Maureen Kane, Deirdre Kelly, Dominic Kelly, Samantha J. L. Knight, Alexandra Y. Kreins, Erika M. Kvikstad, Craig B. Langman, Tracy Lester, Kate E. Lines, Simon R. Lord, Xin Lu, Sahar Mansour, Adnan Manzur, Reza Maroofian, Brian Marsden, Joanne Mason, Simon J. McGowan, Davide Mei, Hana Mlcochova, Yoshiko Murakami, Andrea H. Németh, Steven Okoli, Elizabeth Ormondroyd, Lilian Bomme Ousager, Jacqueline Palace, Smita Y. Patel, Melissa M. Pentony, Chris Pugh, Aboulfazl Rad, Archana Ramesh, Simone G. Riva, Irene Roberts, Noémi Roy, Outi Salminen, Kyleen D. Schilling, Caroline Scott, Arjune Sen, Conrad Smith, Mark Stevenson, Rajesh V. Thakker, Stephen R. F. Twigg, Holm H. Uhlig, Richard van Wijk, Barbara Vona, Steven Wall, Jing Wang, Hugh Watkins, Jaroslav Zak, Anna H. Schuh, Usha Kini, Andrew O. M. Wilkie, Niko Popitsch, and Jenny C. Taylor
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Genome sequencing ,Rare diseases ,Structural variant ,Splice site variant ,Non-coding ,Diagnostic yield ,Medicine ,Genetics ,QH426-470 - Abstract
Abstract Background Whole genome sequencing is increasingly being used for the diagnosis of patients with rare diseases. However, the diagnostic yields of many studies, particularly those conducted in a healthcare setting, are often disappointingly low, at 25–30%. This is in part because although entire genomes are sequenced, analysis is often confined to in silico gene panels or coding regions of the genome. Methods We undertook WGS on a cohort of 122 unrelated rare disease patients and their relatives (300 genomes) who had been pre-screened by gene panels or arrays. Patients were recruited from a broad spectrum of clinical specialties. We applied a bioinformatics pipeline that would allow comprehensive analysis of all variant types. We combined established bioinformatics tools for phenotypic and genomic analysis with our novel algorithms (SVRare, ALTSPLICE and GREEN-DB) to detect and annotate structural, splice site and non-coding variants. Results Our diagnostic yield was 43/122 cases (35%), although 47/122 cases (39%) were considered solved when considering novel candidate genes with supporting functional data into account. Structural, splice site and deep intronic variants contributed to 20/47 (43%) of our solved cases. Five genes that are novel, or were novel at the time of discovery, were identified, whilst a further three genes are putative novel disease genes with evidence of causality. We identified variants of uncertain significance in a further fourteen candidate genes. The phenotypic spectrum associated with RMND1 was expanded to include polymicrogyria. Two patients with secondary findings in FBN1 and KCNQ1 were confirmed to have previously unidentified Marfan and long QT syndromes, respectively, and were referred for further clinical interventions. Clinical diagnoses were changed in six patients and treatment adjustments made for eight individuals, which for five patients was considered life-saving. Conclusions Genome sequencing is increasingly being considered as a first-line genetic test in routine clinical settings and can make a substantial contribution to rapidly identifying a causal aetiology for many patients, shortening their diagnostic odyssey. We have demonstrated that structural, splice site and intronic variants make a significant contribution to diagnostic yield and that comprehensive analysis of the entire genome is essential to maximise the value of clinical genome sequencing.
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- 2023
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4. Generation of human induced pluripotent stem cell line (AOUMEYi001-A) from a patient affected by Congenital disorders of glycosylation (ALG8-CDG) using self-replicating RNA vector
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Rodolfo Tonin, Federica Feo, Silvia Falliano, Lorenzo Ferri, Laura Giunti, Martino Calamai, Elena Procopio, Francesco Mari, Valerio Conti, Ilaria Fanelli, Franco Bambi, Renzo Guerrini, and Amelia Morrone
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Biology (General) ,QH301-705.5 - Abstract
Congenital Disorders of Glycosylation (CDG) are rare inherited metabolic diseases caused by genetic defects in the glycosylation of proteins and lipids. In this study, we describe the generation and characterization of one human induced pluripotent stem cell (hiPSC) line from a 15-year-old male patient with CDG. The patient carried three variants, one (c.122G > A; p.Arg41Gln) inherited from his father and two (c.445 T > G; p.Leu149Arg and the novel c.980C > G; p.Thr327Arg) inherited from his mother in the ALG8 gene (OMIM #608103). The generated hiPSC line shows a normal karyotype, expresses pluripotency markers, and is able to differentiate into the three germ layers.
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- 2023
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5. Circulating tumor cells and palbociclib treatment in patients with ER-positive, HER2-negative advanced breast cancer: results from a translational sub-study of the TREnd trial
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Francesca Galardi, Francesca De Luca, Chiara Biagioni, Ilenia Migliaccio, Giuseppe Curigliano, Alessandro M. Minisini, Martina Bonechi, Erica Moretti, Emanuela Risi, Amelia McCartney, Matteo Benelli, Dario Romagnoli, Silvia Cappadona, Stefano Gabellini, Cristina Guarducci, Valerio Conti, Laura Biganzoli, Angelo Di Leo, and Luca Malorni
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Liquid biopsy ,CTCs ,Luminal breast cancer ,Metastatic ,Palbociclib ,CDK4/6 inhibitor ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Abstract Background Circulating tumor cells (CTCs) are prognostic in patients with advanced breast cancer (ABC). However, no data exist about their use in patients treated with palbociclib. We analyzed the prognostic role of CTC counts in patients enrolled in the cTREnd study, a pre-planned translational sub-study of TREnd (NCT02549430), that randomized patients with ABC to palbociclib alone or palbociclib plus the endocrine therapy received in the prior line of treatment. Moreover, we evaluated RB1 gene expression on CTCs and explored its prognostic role within the cTREnd subpopulation. Methods Forty-six patients with ER-positive, HER2-negative ABC were analyzed. Blood samples were collected before starting palbociclib treatment (timepoint T0), after the first cycle of treatment (timepoint T1), and at disease progression (timepoint T2). CTCs were isolated and counted by CellSearch® System using the CellSearch™Epithelial Cell kit. Progression-free survival (PFS), clinical benefit (CB) during study treatment, and time to treatment failure (TTF) after study treatment were correlated with CTC counts. Samples with ≥ 5 CTCs were sorted by DEPArray system® (DA). RB1 and GAPDH gene expression levels were measured by ddPCR. Results All 46 patients were suitable for CTCs analysis. CTC count at T0 did not show significant prognostic value in terms of PFS and CB. Patients with at least one detectable CTC at T1 (n = 26) had a worse PFS than those with 0 CTCs (n = 16) (p = 0.02). At T1, patients with an increase of at least three CTCs showed reduced PFS compared to those with no increase (mPFS = 3 versus 9 months, (p = 0.004). Finally, patients with ≥ 5 CTCs at T2 (n = 6/23) who received chemotherapy as post-study treatment had a shorter TTF (p = 0.02). Gene expression data for RB1 were obtained from 19 patients. CTCs showed heterogeneous RB1 expression. Patients with detectable expression of RB1 at any timepoint showed better, but not statistically significant, outcomes than those with undetectable levels. Conclusions CTC count seems to be a promising modality in monitoring palbociclib response. Moreover, CTC count at the time of progression could predict clinical outcome post-palbociclib. RB1 expression analysis on CTCs is feasible and may provide additional prognostic information. Results should be interpreted with caution given the small studied sample size.
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- 2021
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6. An Enhanced Distributed Computational Platform for Developmental and Epileptic Encephalopathies.
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Gabriele Cecchetti, Piero Castoldi, Anna Lina Ruscelli, Stefano Dalmiani, Paolo Marcheschi, Federica Pieroni, Davide Galletti, Valerio Conti, and Renzo Guerrini
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- 2022
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7. Developmental and epileptic encephalopathies: from genetic heterogeneity to phenotypic continuum
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Renzo Guerrini, Valerio Conti, Massimo Mantegazza, Simona Balestrini, Aristea S. Galanopoulou, and Fabio Benfenati
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Genetic Heterogeneity ,Epilepsy ,Physiology ,Physiology (medical) ,Mutation ,Animals ,General Medicine ,Molecular Biology - Abstract
Developmental and epileptic encephalopathies (DEEs) are a heterogeneous group of disorders characterized by early-onset, often severe epileptic seizures and EEG abnormalities on a background of developmental impairment that tends to worsen as a consequence of epilepsy. DEEs may result from both nongenetic and genetic etiologies. Genetic DEEs have been associated with mutations in many genes involved in different functions including cell migration, proliferation, and organization, neuronal excitability, and synapse transmission and plasticity. Functional studies performed in different animal models and clinical trials on patients have contributed to elucidate pathophysiological mechanisms underlying many DEEs and have explored the efficacy of different treatments. Here, we provide an extensive review of the phenotypic spectrum included in the DEEs and of the genetic determinants and pathophysiological mechanisms underlying these conditions. We also provide a brief overview of the most effective treatment now available and of the emerging therapeutic approaches.
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- 2023
8. Towards automated neuron tracing via global and local 3D image analysis.
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Ludovica Acciai, Irene Costantini, Francesco Saverio Pavone, Valerio Conti, Renzo Guerrini, Paolo Soda, and Giulio Iannello
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- 2016
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9. Computer-based automatic identification of neurons in gigavoxel-sized 3D human brain images.
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Paolo Soda, Ludovica Acciai, Ermanno Cordelli, Irene Costantini, Leonardo Sacconi, Francesco Saverio Pavone, Valerio Conti, Renzo Guerrini, Paolo Frasconi, and Giulio Iannello
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- 2015
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10. Preliminary Functional Outcomes and Quality of Life after Tongue Reconstruction with the Vastus Lateralis Myofascial Free Flap
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Giuseppe Spriano, Armando De Virgilio, Gerardo Petruzzi, Tiziana Mondello, Jacopo Zocchi, Valerio Conti, Raul Pellini, Andrea Costantino, and Giuseppe Mercante
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medicine.medical_specialty ,Free flap ,Free Tissue Flaps ,Quadriceps Muscle ,03 medical and health sciences ,0302 clinical medicine ,Tongue ,Quality of life ,Interquartile range ,Surveys and Questionnaires ,medicine ,Humans ,Prospective Studies ,030223 otorhinolaryngology ,Prospective cohort study ,Performance status ,business.industry ,Head and neck cancer ,Tongue reconstruction ,Middle Aged ,medicine.disease ,Tongue Neoplasms ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Quality of Life ,Physical therapy ,Surgery ,business - Abstract
The aim of the present study is to report our preliminary experience with the vastus lateralis myofascial free flap (VLMFF) for tongue reconstruction according to tongue and donor site functional outcomes. Twelve consecutive patients (F: 5; median age: 54.0 years, interquartile range or IQR 42.75–69.0) were included. The validated European Organization for Research and Treatment of Cancer of the Head and Neck 35 Quality of Life Questionnaire (EORTC QLQ-H&N35) and the performance status scale for head and neck cancer (PSS-HN) questionnaires were used to assess the health-related quality of life (HRQOL). The lower extremity functional scale (LEFS) was used to self-report the donor area function. All patients were successfully treated, and no VLMFF failure was detected during a median follow-up period of 10.5 months (IQR: 6.5–33.0). The HRQOL showed a median EORTC QLQ-H&N35 score of 56.0 (IQR: 50.0–72.5). The median PSS-HN score was 80.0 (IQR: 45.0–95.0), 75.0 (IQR: 62.5–100.0), 75.0 (IQR: 62.5–100.0) for “Normalcy of Diet,” “Public Eating,” and “Understandability of Speech,” respectively. The self-reported function of the lower extremities (donor area) showed a median LEFS of 59.0 (IQR: 32.5–74.0). This study reports optimistic data regarding the functional and quality of life outcomes after tongue reconstruction using VLMFF. Prospective controlled studies are needed to demonstrate advantages and disadvantages when compared with other reconstructive techniques.
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- 2021
11. Somatic Focal Copy Number Gains of Noncoding Regions of Receptor Tyrosine Kinase Genes in Treatment-Resistant Epilepsy
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Luis Chiriboga, Matija Snuderl, Valerio Conti, Briana Zeck, Javier Hernaez Rodriguez, Adriana Heguy, Maristela L. Onozato, Claudio Forcato, Anna Maria Buccoliero, Hussein Mohamed, Sitharam Ramaswami, Marianna Garonzi, Kaicen Zhu, Jonathan Serrano, Jane A. Skok, Varshini Vasudevaraja, Eveline Teresa Hidalgo, Daniel Friedman, Aristotelis Tsirigos, Arline Faustin, A. John Iafrate, David Zagzag, Renzo Guerrini, Cristiana Pelorosso, James M. Stafford, Orrin Devinsky, Lily M. Tredwin, and John G. Golfinos
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Adult ,Male ,Drug Resistant Epilepsy ,Adolescent ,DNA Copy Number Variations ,PDGFRA ,Receptor tyrosine kinase ,Pathology and Forensic Medicine ,Young Adult ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,Epilepsy ,0302 clinical medicine ,medicine ,Humans ,Epidermal growth factor receptor ,Copy-number variation ,Child ,Gene ,In Situ Hybridization, Fluorescence ,Retrospective Studies ,030304 developmental biology ,0303 health sciences ,biology ,Brain ,Promoter ,Original Articles ,General Medicine ,DNA Methylation ,Middle Aged ,medicine.disease ,ErbB Receptors ,Neurology ,DNA methylation ,Cancer research ,biology.protein ,Female ,Neurology (clinical) ,030217 neurology & neurosurgery - Abstract
Epilepsy is a heterogenous group of disorders defined by recurrent seizure activity due to abnormal synchronized activity of neurons. A growing number of epilepsy cases are believed to be caused by genetic factors and copy number variants (CNV) contribute to up to 5% of epilepsy cases. However, CNVs in epilepsy are usually large deletions or duplications involving multiple neurodevelopmental genes. In patients who underwent seizure focus resection for treatment-resistant epilepsy, whole genome DNA methylation profiling identified 3 main clusters of which one showed strong association with receptor tyrosine kinase (RTK) genes. We identified focal copy number gains involving epidermal growth factor receptor (EGFR) and PDGFRA loci. The dysplastic neurons of cases with amplifications showed marked overexpression of EGFR and PDGFRA, while glial and endothelial cells were negative. Targeted sequencing of regulatory regions and DNA methylation analysis revealed that only enhancer regions of EGFR and gene promoter of PDGFRA were amplified, while coding regions did not show copy number abnormalities or somatic mutations. Somatic focal copy number gains of noncoding regulatory represent a previously unrecognized genetic driver in epilepsy and a mechanism of abnormal activation of RTK genes. Upregulated RTKs provide a potential avenue for therapy in seizure disorders.
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- 2020
12. Profiling PI3K-AKT-MTOR variants in focal brain malformations reveals new insights for diagnostic care
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Filomena Pirozzi, Matthew Berkseth, Rylee Shear, Lorenzo Gonzalez, Andrew E Timms, Josef Sulc, Emily Pao, Nora Oyama, Francesca Forzano, Valerio Conti, Renzo Guerrini, Emily S Doherty, Sulagna C Saitta, Christina M Lockwood, Colin C Pritchard, William B Dobyns, Edward Novotny, Jason N N Wright, Russell P Saneto, Seth Friedman, Jason Hauptman, Jeffrey Ojemann, Raj P Kapur, and Ghayda M Mirzaa
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Original Article ,Neurology (clinical) - Abstract
Focal malformations of cortical development including focal cortical dysplasia, hemimegalencephaly and megalencephaly, are a spectrum of neurodevelopmental disorders associated with brain overgrowth, cellular and architectural dysplasia, intractable epilepsy, autism and intellectual disability. Importantly, focal cortical dysplasia is the most common cause of focal intractable paediatric epilepsy. Gain and loss of function variants in the PI3K-AKT-MTOR pathway have been identified in this spectrum, with variable levels of mosaicism and tissue distribution. In this study, we performed deep molecular profiling of common PI3K-AKT-MTOR pathway variants in surgically resected tissues using droplet digital polymerase chain reaction (ddPCR), combined with analysis of key phenotype data. A total of 159 samples, including 124 brain tissue samples, were collected from 58 children with focal malformations of cortical development. We designed an ultra-sensitive and highly targeted molecular diagnostic panel using ddPCR for six mutational hotspots in three PI3K-AKT-MTOR pathway genes, namely PIK3CA (p.E542K, p.E545K, p.H1047R), AKT3 (p.E17K) and MTOR (p.S2215F, p.S2215Y). We quantified the level of mosaicism across all samples and correlated genotypes with key clinical, neuroimaging and histopathological data. Pathogenic variants were identified in 17 individuals, with an overall molecular solve rate of 29.31%. Variant allele fractions ranged from 0.14 to 22.67% across all mutation-positive samples. Our data show that pathogenic MTOR variants are mostly associated with focal cortical dysplasia, whereas pathogenic PIK3CA variants are more frequent in hemimegalencephaly. Further, the presence of one of these hotspot mutations correlated with earlier onset of epilepsy. However, levels of mosaicism did not correlate with the severity of the cortical malformation by neuroimaging or histopathology. Importantly, we could not identify these mutational hotspots in other types of surgically resected epileptic lesions (e.g. polymicrogyria or mesial temporal sclerosis) suggesting that PI3K-AKT-MTOR mutations are specifically causal in the focal cortical dysplasia-hemimegalencephaly spectrum. Finally, our data suggest that ultra-sensitive molecular profiling of the most common PI3K-AKT-MTOR mutations by targeted sequencing droplet digital polymerase chain reaction is an effective molecular approach for these disorders with a good diagnostic yield when paired with neuroimaging and histopathology.
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- 2022
13. Atypical Ocular Coloboma in Tuberous Sclerosis-2: Report of Two Novel Cases
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Roberto Caputo, Renzo Guerrini, Valerio Conti, Giacomo Bacci, Francesco Mari, and Silvio Polizzi
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Male ,Fovea Centralis ,congenital, hereditary, and neonatal diseases and abnormalities ,medicine.medical_specialty ,DNA Mutational Analysis ,Visual Acuity ,Iris ,Retina ,Tuberous sclerosis ,chemistry.chemical_compound ,Tuberous Sclerosis ,Tuberous Sclerosis Complex 2 Protein ,Humans ,Medicine ,Abnormalities, Multiple ,Papilledema ,business.industry ,Retinal ,DNA ,medicine.disease ,Dermatology ,Iris coloboma ,Angiofibromas ,eye diseases ,nervous system diseases ,Coloboma ,Ophthalmology ,medicine.anatomical_structure ,chemistry ,Child, Preschool ,Mutation ,Female ,sense organs ,Neurology (clinical) ,Eyelid ,TSC1 ,medicine.symptom ,TSC2 ,business ,Tomography, Optical Coherence - Abstract
Tuberous sclerosis complex (TSC) is an autosomal dominant multisystemic disorder caused by mutations in either TSC1 or TSC2 genes and is characterized by hamartomas in multiple organs. The most frequent and best-known ocular manifestation in TSC is the retinal hamartoma. Less frequent ocular manifestations include punched out areas of retinal depigmentation, eyelid angiofibromas, uveal colobomas, papilledema, and sector iris depigmentation. In this article, we report 2 patients carrying known pathogenic variants in the TSC2 gene who exhibited an atypical, unilateral, iris coloboma associated with localized areas of retinal dysembryogenesis.
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- 2020
14. <scp>Pre‐Clinical</scp> Experience With the <scp>VITOM 3D</scp> and the <scp>ARTip</scp> Cruise System for <scp>Micro‐Laryngeal</scp> Surgery
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Tiziana Mondello, Elena Russo, Giuseppe Mercante, Armando De Virgilio, Andrea Costantino, Giuseppe Spriano, Valerio Conti, and Francesca Pirola
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Larynx ,medicine.medical_specialty ,Laryngoscopy ,business.industry ,General surgery ,Cruise ,MEDLINE ,Pilot Projects ,Equipment Design ,Robotics ,Laryngoscopes ,medicine.anatomical_structure ,Otorhinolaryngology ,Surgical Procedures, Operative ,medicine ,Robotic surgery ,Prospective Studies ,Laryngeal surgery ,business - Published
- 2020
15. Genomic DNA methylation distinguishes subtypes of human focal cortical dysplasia
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Roland Coras, Ingmar Blümcke, Renzo Guerrini, Tom Pieper, Hans Holthausen, Eleonora Aronica, Johannes A. Hainfellner, Valerio Conti, Assam El-Osta, Harikrishnan Kaipananickal, Antony Kaspi, Manfred Kudernatsch, Martha Feucht, Thomas Czech, Hajo M. Hamer, Mark Ziemann, Karl Rössler, Angelika Mühlebner, Ishant Khurana, Burkhard S. Kasper, Katja Kobow, Pathology, ANS - Cellular & Molecular Mechanisms, APH - Aging & Later Life, and APH - Mental Health
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Adult ,Male ,focal epilepsy ,0301 basic medicine ,diagnostic biomarker ,Adolescent ,Tissue Banks ,Biology ,Young Adult ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Cluster Analysis ,Humans ,RNA, Messenger ,Epigenetics ,Child ,Gene ,Tomography, Emission-Computed, Single-Photon ,Massive parallel sequencing ,Genome, Human ,Gene Expression Profiling ,TOR Serine-Threonine Kinases ,Infant ,DNA ,Methylation ,DNA Methylation ,Middle Aged ,Cortical dysplasia ,medicine.disease ,DEPDC5 ,3. Good health ,Malformations of Cortical Development ,genomic DNA ,mTOR pathway ,030104 developmental biology ,Neurology ,Child, Preschool ,DNA methylation ,Full‐length Original Research ,Cancer research ,Female ,Epilepsies, Partial ,Neurology (clinical) ,Tomography, X-Ray Computed ,Transcriptome ,focal cortical dysplasia ,epigenetic ,030217 neurology & neurosurgery - Abstract
Objectives Focal cortical dysplasia (FCD) is a major cause of drug‐resistant focal epilepsy in children, and the clinicopathological classification remains a challenging issue in daily practice. With the recent progress in DNA methylation–based classification of human brain tumors we examined whether genomic DNA methylation and gene expression analysis can be used to also distinguish human FCD subtypes. Methods DNA methylomes and transcriptomes were generated from massive parallel sequencing in 15 surgical FCD specimens, matched with 5 epilepsy and 6 nonepilepsy controls. Results Differential hierarchical cluster analysis of DNA methylation distinguished major FCD subtypes (ie, Ia, IIa, and IIb) from patients with temporal lobe epilepsy patients and nonepileptic controls. Targeted panel sequencing identified a novel likely pathogenic variant in DEPDC5 in a patient with FCD type IIa. However, no enrichment of differential DNA methylation or gene expression was observed in mechanistic target of rapamycin (mTOR) pathway–related genes. Significance Our studies extend the evidence for disease‐specific methylation signatures toward focal epilepsies in favor of an integrated clinicopathologic and molecular classification system of FCD subtypes incorporating genomic methylation.
- Published
- 2019
16. Circulating tumor cells and palbociclib treatment in patients with ER-positive, HER2-negative advanced breast cancer: results from a translational sub-study of the TREnd trial
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Stefano Gabellini, Angelo Di Leo, Erica Moretti, Giuseppe Curigliano, Luca Malorni, C. Guarducci, Francesca De Luca, Matteo Benelli, Ilenia Migliaccio, Emanuela Risi, Chiara Biagioni, Laura Biganzoli, Martina Bonechi, Amelia McCartney, Silvia Cappadona, Dario Romagnoli, Francesca Galardi, Valerio Conti, and Alessandro Marco Minisini
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Oncology ,medicine.medical_specialty ,Pyridines ,Receptor, ErbB-2 ,Ubiquitin-Protein Ligases ,medicine.medical_treatment ,ddPCR ,Breast Neoplasms ,Cell Count ,Palbociclib ,lcsh:RC254-282 ,Piperazines ,CDK4/6 inhibitor ,03 medical and health sciences ,0302 clinical medicine ,Circulating tumor cell ,Breast cancer ,Luminal breast cancer ,Surgical oncology ,Internal medicine ,Biomarkers, Tumor ,Humans ,Medicine ,Liquid biopsy ,Protein Kinase Inhibitors ,030304 developmental biology ,0303 health sciences ,Chemotherapy ,business.industry ,Cancer ,Biomarker ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Neoplastic Cells, Circulating ,medicine.disease ,Progression-Free Survival ,Retinoblastoma Binding Proteins ,Treatment Outcome ,Receptors, Estrogen ,030220 oncology & carcinogenesis ,Disease Progression ,Metastatic ,Biomarker (medicine) ,Female ,CTCs ,Receptors, Progesterone ,business ,Research Article - Abstract
Background Circulating tumor cells (CTCs) are prognostic in patients with advanced breast cancer (ABC). However, no data exist about their use in patients treated with palbociclib. We analyzed the prognostic role of CTC counts in patients enrolled in the cTREnd study, a pre-planned translational sub-study of TREnd (NCT02549430), that randomized patients with ABC to palbociclib alone or palbociclib plus the endocrine therapy received in the prior line of treatment. Moreover, we evaluated RB1 gene expression on CTCs and explored its prognostic role within the cTREnd subpopulation. Methods Forty-six patients with ER-positive, HER2-negative ABC were analyzed. Blood samples were collected before starting palbociclib treatment (timepoint T0), after the first cycle of treatment (timepoint T1), and at disease progression (timepoint T2). CTCs were isolated and counted by CellSearch® System using the CellSearch™Epithelial Cell kit. Progression-free survival (PFS), clinical benefit (CB) during study treatment, and time to treatment failure (TTF) after study treatment were correlated with CTC counts. Samples with ≥ 5 CTCs were sorted by DEPArray system® (DA). RB1 and GAPDH gene expression levels were measured by ddPCR. Results All 46 patients were suitable for CTCs analysis. CTC count at T0 did not show significant prognostic value in terms of PFS and CB. Patients with at least one detectable CTC at T1 (n = 26) had a worse PFS than those with 0 CTCs (n = 16) (p = 0.02). At T1, patients with an increase of at least three CTCs showed reduced PFS compared to those with no increase (mPFS = 3 versus 9 months, (p = 0.004). Finally, patients with ≥ 5 CTCs at T2 (n = 6/23) who received chemotherapy as post-study treatment had a shorter TTF (p = 0.02). Gene expression data for RB1 were obtained from 19 patients. CTCs showed heterogeneous RB1 expression. Patients with detectable expression of RB1 at any timepoint showed better, but not statistically significant, outcomes than those with undetectable levels. Conclusions CTC count seems to be a promising modality in monitoring palbociclib response. Moreover, CTC count at the time of progression could predict clinical outcome post-palbociclib. RB1 expression analysis on CTCs is feasible and may provide additional prognostic information. Results should be interpreted with caution given the small studied sample size.
- Published
- 2021
17. Angiocentric glioma-associated seizures: The possible role of EATT2, pyruvate carboxylase and glutamine synthetase
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Sabrina Rossi, Felice Giangaspero, Anna Maria Buccoliero, Marco Gessi, Lorenzo Genitori, Mariarita Santi, Flavio Giordano, Luca Bertero, Eleonora Aronica, Mirko Scagnet, Selene Moscardi, Vittoria Donofrio, Federico Mussa, Carmen Barba, Valerio Conti, Irene Migliastro, Chiara Caporalini, Francesca Gianno, Francesca Diomedi-Camassei, Renzo Guerrini, Iacopo Sardi, Manila Antonelli, Pathology, APH - Aging & Later Life, APH - Mental Health, and Amsterdam Neuroscience - Cellular & Molecular Mechanisms
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LEAT ,Angiocentric Glioma ,Glutamic Acid ,Pathogenesis ,03 medical and health sciences ,Epilepsy ,Brain ,Central nervous system ,Tumor ,Glutamate-Ammonia Ligase ,Humans ,Pyruvate Carboxylase ,Glioma ,Seizures ,0302 clinical medicine ,Glutamine synthetase ,medicine ,Chemistry ,Glutamate receptor ,General Medicine ,Glutamic acid ,medicine.disease ,Pyruvate carboxylase ,Neurology ,Cancer research ,Neurology (clinical) ,030217 neurology & neurosurgery - Abstract
Purpose Our purpose was to better understand the pathogenesis of seizures associated with angiocentric glioma. Angiocentric glioma is an indolent and rare low-grade glioma. Its typical clinical presentation is with epileptic seizures. The pathogenesis of tumor-associated seizures is poorly understood. Among the possible pathomechanisms, the increased neurotoxic concentrations of the glutamate has been proposed. Glutamate transporters, pyruvate carboxylase and glutamine synthetase are involved in maintaining the physiological concentration of glutamate in the inter synaptic spaces. Methods We evaluated the immunohistochemical expression of EAAT2 (the most important glutamate transporter), pyruvate carboxylase and glutamine synthetase in 17 angiocentric gliomas. Results EAAT2 was never expressed (0%) in the neoplastic cells in none of the cases studied. Pyruvate carboxylase was expressed in the cytoplasm of the neoplastic cells in 16/17 cases (94 %). Glutamine synthetase was expressed in the cytoplasm of the neoplastic cells in 15/17 cases (88 %). Conclusion The net result of this enzymatic expression, in particular considering the loss of EAAT2, could be an increased glutamate concentration in the synaptic clef, which might increase local network excitability initially involving intratumoral neurons. The observation that the angiocentric glioma-associated epilepsy might be at least in part related to EAAT2 deficiency opens up interesting therapeutic perspectives.
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- 2021
18. Prospective Evaluation of Ghrelin and Des-Acyl Ghrelin Plasma Levels in Children with Newly Diagnosed Epilepsy: Evidence for Reduced Ghrelin-to-Des-Acyl Ghrelin Ratio in Generalized Epilepsies
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Anna-Maria Costa, Tommaso Lo Barco, Elisabetta Spezia, Valerio Conti, Laura Roli, Lorenza Marini, Sara Minghetti, Elisa Caramaschi, Laura Pietrangelo, Luca Pecoraro, Fabio D’Achille, Paola Accorsi, Tommaso Trenti, Federico Melani, Carla Marini, Renzo Guerrini, Francesca Darra, Patrizia Bergonzini, and Giuseppe Biagini
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children ,ghrelin ,digestive, oral, and skin physiology ,epilepsy ,Medicine (miscellaneous) ,generalized epilepsy ,antiseizure medications ,plasma - Abstract
Children with epilepsy and identified as responders to antiseizure medications (ASMs) were found to present markedly higher ghrelin plasma levels when compared to drug-resistant patients. However, it was undetermined if this phenotype could be influenced by the ASMs. Here, we prospectively investigated total ghrelin and des-acyl ghrelin (DAG) plasma levels by enzyme-linked immunosorbent assay before and after ASM administration. Inclusion criteria were: (i) subject with a suspicion of epilepsy; (ii) age ranging from 0 to 16 years; and (iii) informed consent signed by parents or caregivers. Exclusion criteria were acute or chronic metabolic disorders with occasional convulsions but without epilepsy. Fifty patients were followed over a period of one year in Italian neuropediatric centers. Apart from a few exceptions, the majority of children were responsive to ASMs. No differences were found in total ghrelin and DAG levels before and after the treatment, but total ghrelin levels were significantly lower in children with generalized epilepsy compared to those with combined focal and generalized epilepsy. Moreover, the ghrelin-to-DAG ratio was also markedly lower in generalized epilepsies compared to all the other types of epilepsy. Finally, ghrelin was unchanged by ASMs, including the first (e.g., carbamazepine), second (levetiracetam), and third (lacosamide) generation of anticonvulsants.
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- 2022
19. A combined pipeline for quantitative analysis of human brain cytoarchitecture
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Christophe Destrieux, Luca Pesce, Francesco S. Pavone, Andrea Simonetto, Annunziatina Laurino, Renzo Guerrini, Erica Lazzeri, Giovanni Lughi, Mattia Neri, Irene Costantini, Filippo Maria Castelli, Ludovico Silvestri, Matteo Roffilli, Valerio Conti, and Giacomo Mazzamuto
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Structural organization ,business.industry ,Computer science ,3d analysis ,Pipeline (computing) ,Pattern recognition ,Human brain ,Convolutional neural network ,medicine.anatomical_structure ,Cytoarchitecture ,Cortex (anatomy) ,medicine ,Artificial intelligence ,business - Abstract
The 3D analysis of the human brain architecture at cellular resolution is still a big challenge. In this work, we propose a pipeline that solves the problem of performing neuronal mapping in large human brain samples at micrometer resolution. First, we introduce the SWITCH/TDE protocol: a robust methodology to clear and label human brain tissue. Then, we implement the 2.5D method based on a Convolutional Neural Network, to automatically detect and segment all neurons. Our method proved to be highly versatile and was applied successfully on specimens from different areas of the cortex originating from different subjects (young, adult and elderly, both healthy and pathological). We quantitatively evaluate the density and, more importantly, the mean volume of the thousands of neurons identified within the specimens. In conclusion, our pipeline makes it possible to study the structural organization of the brain and expands the histopathological studies to the third dimension.
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- 2020
20. Is Focal Cortical Dysplasia/Epilepsy Caused by Somatic
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Renzo, Guerrini, Mara, Cavallin, Tommaso, Pippucci, Anna, Rosati, Francesca, Bisulli, Paola, Dimartino, Carmen, Barba, Rita, Garbelli, Anna Maria, Buccoliero, Laura, Tassi, and Valerio, Conti
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Views & Reviews - Abstract
Objective To alert about the wide margin of unpredictability that distribution of somatic MTOR mosaicism may have in the brain and the risk for independent epileptogenesis arising from the seemingly healthy contralateral hemisphere after complete removal of epileptogenic focal cortical dysplasia (FCD). Methods Clinical, EEG, MRI, histopathology, and molecular genetics in 2 patients (1 and 2) treated with focal resections and subsequent complete hemispherectomy for epileptogenic FCD due to somatic MTOR mutations. Autoptic brain study of bilateral asymmetric hemispheric dysplasia and identification of alternative allele fraction (AAF) rates for AKT1 (patient 3). Results The strongly hyperactivating p.Ser2215Phe (patient 1) and p.Leu1460Pro (patient 2) MTOR mutations were at low-level AAF in the dysplastic tissue. After repeated resections and eventual complete hemispherectomy, both patients manifested intractable seizures arising from the contralateral, seemingly healthy hemisphere. In patient 3, the p.Glu17Lys AKT1 mutation exhibited random distribution and AAF rates in different tissues with double levels in the more severely dysplastic cerebral hemisphere. Conclusions Our understanding of the distribution of somatic mutations in the brain in relation to the type of malformation and its hypothesized time of origin may be faulty. Large studies may reveal that the risk of a first surgery being disappointing might be related more to the specific somatic mammalian target of rapamycin mutation identified than to completeness of resection and that the advantages of repeated resections after a first unsuccessful operation should be weighed against the risk of the contralateral hemisphere becoming in turn epileptogenic.
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- 2020
21. High definition three-dimensional exoscope (VITOM 3D) for microsurgery training: a preliminary experience
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Valerio Conti, Giovanni Cugini, Andrea Costantino, Tiziana Mondello, Matteo Di Bari, Giuseppe Spriano, Claudia Ebm, Armando De Virgilio, and Giuseppe Mercante
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medicine.medical_specialty ,Microsurgery ,business.industry ,Quality assessment ,medicine.medical_treatment ,Significant difference ,General Medicine ,Likert scale ,03 medical and health sciences ,0302 clinical medicine ,System quality ,Otorhinolaryngology ,030220 oncology & carcinogenesis ,Cohort ,Physical therapy ,Medicine ,High definition ,Humans ,030223 otorhinolaryngology ,business - Abstract
To assess the feasibility of a high definition 3D exoscope (VITOM®) for microsurgery training in a cohort of naive medical students. Twenty-two consecutive medical students performed a battery of four exercises assessing basic microsurgical skills. The students were randomized in two different groups based on two different VITOM® holding systems (VERSACRANE™ and ARTip™ cruise). Participants self-reported the VITOM® system quality on a 4-point Likert scale (VITOM Quality Assessment Tool). The time needed to complete the exercises was analyzed. All students successfully completed the training, and no technical issues were raised during the simulation. The majority of the individual items were judged “good” or “very good” (n = 187; 94.4%), regardless of the two groups. “Image quality” (n = 21; 95%), “magnification rate” (n = 20; 91%), “stereoscopic effect” (n = 19; 86%), and “focusing” (n = 18; 82%) represented the best-rated items. No statistically significant difference between the two groups was measured in almost all items of the VITOM Quality Assessment Tool (p > 0.05). The time needed to perform each exercise showed a statistically significant difference between groups in two tests (p
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- 2020
22. International consensus recommendations on the diagnostic work-up for malformations of cortical development
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Daniela T. Pilz, Katrien Stouffs, Orly Reiner, Anna Jansen, Eleonora Aronica, Edith Said, Renske Oegema, WB Dobyns, Richard J. Leventer, David Gomez Andres, Nadia Bahi-Buisson, Martina Wilke, Ivana Pogledic, Dina Amrom, Ute Hehr, Ghayda Mirzaa, Elena Parrini, Luis M Valor, Renzo Guerrini, Valerio Conti, Andrew E. Fry, Doriette Soler, Tahsin Stefan Barakat, Grazia M.S. Mancini, Nataliya Di Donato, Maha S. Zaki, T. Geis, Clinical Genetics, Clinical sciences, Medical Genetics, Reproduction and Genetics, Physiotherapy, Human Physiology and Anatomy, Pediatrics, Public Health Sciences, Mental Health and Wellbeing research group, Neurogenetics, Pathology, APH - Aging & Later Life, APH - Mental Health, and ANS - Cellular & Molecular Mechanisms
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medicine.medical_specialty ,Consensus ,Internationality ,Delphi Technique ,MEDLINE ,Cerebral palsy ,Cellular and Molecular Neuroscience ,Epilepsy ,Neurologie ,Intellectual disability ,medicine ,Humans ,Epilepsy surgery ,Intensive care medicine ,Diagnostic Tests, Routine ,business.industry ,Neurodevelopmental disorders ,Consensus Statement ,medicine.disease ,Work-up ,Paediatric neurological disorders ,Malformations of Cortical Development ,Workflow ,Practice Guidelines as Topic ,Etiology ,Neurology (clinical) ,business ,Sciences cognitives - Abstract
Malformations of cortical development (MCDs) are neurodevelopmental disorders that result from abnormal development of the cerebral cortex in utero. MCDs place a substantial burden on affected individuals, their families and societies worldwide, as these individuals can experience lifelong drug-resistant epilepsy, cerebral palsy, feeding difficulties, intellectual disability and other neurological and behavioural anomalies. The diagnostic pathway for MCDs is complex owing to wide variations in presentation and aetiology, thereby hampering timely and adequate management. In this article, the international MCD network Neuro-MIG provides consensus recommendations to aid both expert and non-expert clinicians in the diagnostic work-up of MCDs with the aim of improving patient management worldwide. We reviewed the literature on clinical presentation, aetiology and diagnostic approaches for the main MCD subtypes and collected data on current practices and recommendations from clinicians and diagnostic laboratories within Neuro-MIG. We reached consensus by 42 professionals from 20 countries, using expert discussions and a Delphi consensus process. We present a diagnostic workflow that can be applied to any individual with MCD and a comprehensive list of MCD-related genes with their associated phenotypes. The workflow is designed to maximize the diagnostic yield and increase the number of patients receiving personalized care and counselling on prognosis and recurrence risk., SCOPUS: ar.j, DecretOANoAutActif, info:eu-repo/semantics/published
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- 2020
23. Corrigendum to ‘Angiocentric glioma-associated seizures: The possible role of EATT2, pyruvate carboxylase and glutamine synthetase [Seizure: European Journal of Epilepsy 86 (2021) 152-154]
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Sabrina Rossi, Valerio Conti, Francesca Diomedi-Camassei, Manila Antonelli, Flavio Giordano, Carmen Barba, Luca Bertero, Anna Maria Buccoliero, Lorenzo Genitori, Federico Mussa, Vittoria Donofrio, Iacopo Sardi, Eleonora Aronica, Felice Giangaspero, Irene Migliastro, Selene Moscardi, Mariarita Santi, Marco Gessi, Renzo Guerrini, Chiara Caporalini, Francesca Gianno, and Mirko Scagnet
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Epilepsy ,Neurology ,Angiocentric Glioma ,Glutamine synthetase ,Cancer research ,medicine ,Neurology (clinical) ,General Medicine ,Biology ,medicine.disease ,Pyruvate carboxylase - Published
- 2021
24. Multimodal fiber-probe spectroscopy allows detecting epileptogenic focal cortical dysplasia in children
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Valerio Conti, Anna Maria Buccoliero, Francesco S. Pavone, Suresh Anand, Renzo Guerrini, Riccardo Cicchi, and Flavio Giordano
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Surgical resection ,Pathology ,medicine.medical_specialty ,Materials science ,General Physics and Astronomy ,Brain tissue ,Spectrum Analysis, Raman ,Multimodal Imaging ,01 natural sciences ,General Biochemistry, Genetics and Molecular Biology ,Fluorescence spectroscopy ,010309 optics ,03 medical and health sciences ,symbols.namesake ,0302 clinical medicine ,Nuclear magnetic resonance ,0103 physical sciences ,medicine ,Humans ,General Materials Science ,Child ,Spectroscopy ,Principal Component Analysis ,General Engineering ,General Chemistry ,Cortical dysplasia ,medicine.disease ,Malformations of Cortical Development ,Spectrometry, Fluorescence ,symbols ,Fiber probe ,Raman spectroscopy ,030217 neurology & neurosurgery - Abstract
Schematic draw of the experimental setup used for fiber-probe spectroscopy. We evaluated the diagnostic capability of a multimodal spectroscopic approach for classifying normal brain tissue and epileptogenic focal cortical dysplasia in children. We employed fluorescence spectroscopy at two excitation wavelengths (378 nm and 445 nm) and Raman spectroscopy (at 785 nm excitation) for acquiring fluorescence and Raman spectra from 10 normal brains, 16 focal cortical dysplasia specimens and 1 cortical tuber tissue sites using a custom-built multimodal optical point spectroscopic system. We used principal component analysis combined with leave-one-sample-out-cross-validation for tissue classification. The study resulted in 100% sensitivity and 90% specificity using the information obtained from fluorescence at two distinct wavelengths and Raman spectroscopy for discriminating normal brain tissue and focal cortical dysplasia. Our results demonstrate that this methodology has the potential to be applied clinically for the detection of focal cortical dysplasia and can help to improve as precise as possible surgical resection of the dysplastic tissue during surgery for epilepsy.
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- 2017
25. Lesional and non-lesional epilepsies: A blurring genetic boundary
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Elena Parrini, Alessandro Esposito, Renzo Guerrini, Anna Fassio, and Valerio Conti
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Epileptogenic structural abnormalities ,Genetic epilepsies ,Neurodevelopmental disorders with epilepsy ,Phenotypic and genetic heterogeneity ,Epilepsy ,Developmental Disabilities ,Boundary (topology) ,General Medicine ,Biology ,Epileptogenesis ,03 medical and health sciences ,0302 clinical medicine ,Neuroimaging ,030225 pediatrics ,Pediatrics, Perinatology and Child Health ,Humans ,Neurology (clinical) ,Neuroscience ,030217 neurology & neurosurgery - Abstract
There has been a traditional conceptual partition between the so-called non-lesional genetic epilepsies and the genetically determined interposed epileptogenic structural abnormalities. In this review, we summarise how growing evidence acquired through neuroimaging and neurobiology modelling is demonstrating that a distinction between lesional and functional (or non-lesional) epileptogenesis is less obvious than previously thought, particularly for epileptogenic neurodevelopmental disorders, but also for most genetically determined epilepsies.
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- 2019
26. Correction: The landscape of epilepsy-related GATOR1 variants
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Johannes R. Lemke, Thomas Dorn, Laura Hernandez-Hernandez, Giuseppe d'Orsi, Eveline Hagebeuk, Annapurna Poduri, Douglas R. NordliJr, Hélène Catenoix, Melanie Jennesson, Veronique Darmency, Kevin Rostasy, Thomas Becher, Laura Licchetta, Peter Uldall, Lysa Boissé Lomax, Bianca Berghuis, Ilona Krey, Gaetan Lesca, Anne de Saint Martin, Boudewijn Gunning, Anne Fabienne Lepine, Renske Oegema, Brigitte Ricard-Mousnier, Sarah Ferrand-Sorbets, Arnaud Biraben, Charles Deckers, An Sofie Schoonjans, Martin Zenker, Simona Balestrini, Inga Talvik, Julitta de Bellescize, Christopher J. Yuskaitis, Georg Dorfmüller, Erik H. Niks, Guillaume Achaz, Laurence Faivre, Pasquale Striano, Shifteh Sattar, Karen Müller-Schlüter, Sanjay M. Sisodiya, Caroline Nava, Pia Zacher, Fabrice Bartolomei, Kristin Lindstrom, Sophie Julia, Eric LeGuern, Thomas Cloppenborg, Rikke S. Møller, Antonio Gambardella, Mathilde Chipaux, Sara Baldassari, Pavel Krsek, Marie Line Jacquemont, Ilse Wegner, Christian Brandt, Barbora Benova, Valerio Conti, Gabrielle Rudolf, Floor E. Jansen, Edouard Hirsch, S. Krithika, Jamel Chelly, Katrien Stouffs, Daniëlle de Jong, Courtney Kiss, Tilman Polster, Eva H. Brilstra, Tommaso Pippucci, Markus Wolff, Cécile Marchal, Giovanni Crichiutti, Kees P.J. Braun, Paolo Tinuper, Stéphanie Baulac, Valentin Sander, Anne-Sophie Lebre, Nienke E. Verbeek, Anna Jansen, Francesca Bisulli, Fabienne Picard, Natasha E. Schoeler, Julien Thevenon, Marjan J. A. van Kempen, Guido Rubboli, Sarah Weckhuysen, Renzo Guerrini, Eleni Panagiotakaki, Meral Balci, Institut du Cerveau et de la Moëlle Epinière = Brain and Spine Institute (ICM), Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Hôpital Universitaire de Genève, Department of Genetics [Utrecht, the Netherlands], University Medical Center [Utrecht], Children's Hospital A. Meyer, Department of Clinical and Experimental Epilepsy, University College of London [London] (UCL), Molecular Cell Biology, Leiden University Medical Center (LUMC), Danish Epilepsy Centre, Denmark and Aarhus University, Aarhus, Centre Hospitalier Universitaire de La Réunion (CHU La Réunion), Service de Génétique Cytogénétique et Embryologie [CHU Pitié-Salpêtrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Neurogénétique Moléculaire et Cellulaire, Institut Naltional de la Santé et de la Recherche Médicale, Epidémiologie et analyses en santé publique : risques, maladies chroniques et handicaps (LEASP), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Regional Epilepsy Center, Reggio Calabria, Centre de génétique - Centre de référence des maladies rares, anomalies du développement et syndromes malformatifs (CHU de Dijon), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), FHU TRANSLAD (CHU de Dijon), Département de Sociologie [université de Bourgogne], UFR de sciences humaines [Université de Bourgogne], Université de Bourgogne (UB)-Université de Bourgogne (UB), Service de Neurophysiologie Clinique (CHU Dijon), Laboratoire Traitement du Signal et de l'Image (LTSI), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de génétique [Reims], Centre Hospitalier Universitaire de Reims (CHU Reims), Universitair Ziekenhuis Brussel, Equipe GAD (LNC - U1231), Lipides - Nutrition - Cancer [Dijon - U1231] (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement, Université Bourgogne Franche-Comté [COMUE] (UBFC), Département d'Epilepsie, Sommeil et Neurophysiologie Pédiatrique [HCL, Lyon], Hospices Civils de Lyon (HCL), Institute of Human Genetics, University Hospital Magdeburg, Institut de Systématique, Evolution, Biodiversité (ISYEB ), Muséum national d'Histoire naturelle (MNHN)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), University of Southern Denmark (SDU), Institut du Cerveau = Paris Brain Institute (ICM), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Hôpital Universitaire de Genève = University Hospitals of Geneva (HUG), Universiteit Leiden-Universiteit Leiden, Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Muséum national d'Histoire naturelle (MNHN)-École Pratique des Hautes Études (EPHE), Centre interdisciplinaire de recherche en biologie (CIRB), Labex MemoLife, École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Collège de France (CdF (institution))-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)
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0303 health sciences ,business.industry ,Published Erratum ,MEDLINE ,medicine.disease ,computer.software_genre ,Spelling ,03 medical and health sciences ,Epilepsy ,0302 clinical medicine ,[SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics ,medicine ,Artificial intelligence ,business ,Psychology ,computer ,030217 neurology & neurosurgery ,Genetics (clinical) ,Natural language processing ,030304 developmental biology - Abstract
International audience; The original version of this article contained an error in the spelling of the author Erik H. Niks, which was incorrectly given as Erik Niks. This has now been corrected in both the PDF and HTML versions of the article.
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- 2019
27. Biallelic DMXL2 mutations impair autophagy and cause Ohtahara syndrome with progressive course
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Matias Wagner, Hava Hafner, Erez Y. Levanon, Alessandro Esposito, Antonio Falace, Silvia Giovedì, Antonio De Fusco, Tiziana Pisano, Sarit Ravid, Hanna Mandel, Davide Aprile, Ayelet Eran, Sarah L. Stenton, Saskia B. Wortmann, Annette Seibt, Maria Sabina Cerullo, Davide Mei, Valerio Conti, Daniella Magen, Carla Marini, Chiara Fiorillo, Felix Distelmaier, Ertan Mayatepek, Fabio Benfenati, Tilman Polster, Moran Gal, Anna Fassio, and Renzo Guerrini
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0301 basic medicine ,Male ,medicine.medical_specialty ,Ohtahara syndrome ,Synaptogenesis ,Nerve Tissue Proteins ,Compound heterozygosity ,autophagy ,developmental and epileptic encephalopathy ,neuropathy ,progressive disorder ,Leukoencephalopathy ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,Exome Sequencing ,Autophagy ,Medicine ,Missense mutation ,Humans ,Child ,Developmental And Epileptic Encephalopathy ,Neuropathy ,Ohtahara Syndrome ,Progressive Disorder ,Loss function ,Exome sequencing ,Adaptor Proteins, Signal Transducing ,business.industry ,Brain ,Infant ,Electroencephalography ,medicine.disease ,Magnetic Resonance Imaging ,Pedigree ,030104 developmental biology ,Endocrinology ,Child, Preschool ,Mutation ,Disease Progression ,Female ,Neurology (clinical) ,business ,Lysosomes ,Spasms, Infantile ,030217 neurology & neurosurgery - Abstract
Ohtahara syndrome, early infantile epileptic encephalopathy with a suppression burst EEG pattern, is an aetiologically heterogeneous condition starting in the first weeks or months of life with intractable seizures and profound developmental disability. Using whole exome sequencing, we identified biallelic DMXL2 mutations in three sibling pairs with Ohtahara syndrome, belonging to three unrelated families. Siblings in Family 1 were compound heterozygous for the c.5135C>T (p.Ala1712Val) missense substitution and the c.4478C>G (p.Ser1493*) nonsense substitution; in Family 2 were homozygous for the c.4478C>A (p.Ser1493*) nonsense substitution and in Family 3 were homozygous for the c.7518-1G>A (p.Trp2507Argfs*4) substitution. The severe developmental and epileptic encephalopathy manifested from the first day of life and was associated with deafness, mild peripheral polyneuropathy and dysmorphic features. Early brain MRI investigations in the first months of life revealed thin corpus callosum with brain hypomyelination in all. Follow-up MRI scans in three patients revealed progressive moderate brain shrinkage with leukoencephalopathy. Five patients died within the first 9 years of life and none achieved developmental, communicative or motor skills following birth. These clinical findings are consistent with a developmental brain disorder that begins in the prenatal brain, prevents neural connections from reaching the expected stages at birth, and follows a progressive course. DMXL2 is highly expressed in the brain and at synaptic terminals, regulates v-ATPase assembly and activity and participates in intracellular signalling pathways; however, its functional role is far from complete elucidation. Expression analysis in patient-derived skin fibroblasts demonstrated absence of the DMXL2 protein, revealing a loss of function phenotype. Patients' fibroblasts also exhibited an increased LysoTracker® signal associated with decreased endolysosomal markers and degradative processes. Defective endolysosomal homeostasis was accompanied by impaired autophagy, revealed by lower LC3II signal, accumulation of polyubiquitinated proteins, and autophagy receptor p62, with morphological alterations of the autolysosomal structures on electron microscopy. Altered lysosomal homeostasis and defective autophagy were recapitulated in Dmxl2-silenced mouse hippocampal neurons, which exhibited impaired neurite elongation and synaptic loss. Impaired lysosomal function and autophagy caused by biallelic DMXL2 mutations affect neuronal development and synapse formation and result in Ohtahara syndrome with profound developmental impairment and reduced life expectancy.
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- 2019
28. Multiple genomic copy number variants associated with periventricular nodular heterotopia indicate extreme genetic heterogeneity
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Sebastiano A. Musumeci, Elena Parrini, Carla Marini, Renzo Guerrini, Elena Cellini, Claudia Clementella, Matteo Della Monica, Viola Doccini, Marco Fichera, Annalisa Vetro, Valerio Conti, and Sabrina Giglio
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Proband ,Male ,DNA Copy Number Variations ,Genetics ,Genetics (clinical) ,Biology ,Article ,03 medical and health sciences ,Lateral ventricles ,Genetic Heterogeneity ,0302 clinical medicine ,Periventricular Nodular Heterotopia ,hemic and lymphatic diseases ,Polymicrogyria ,medicine ,FLNA ,Humans ,Copy-number variation ,Gene ,030304 developmental biology ,Chromosome Aberrations ,0303 health sciences ,Genetic heterogeneity ,Breakpoint ,medicine.disease ,Magnetic Resonance Imaging ,3. Good health ,Female ,030217 neurology & neurosurgery - Abstract
Periventricular nodular heterotopia (PNH) is a brain malformation in which nodules of neurons are ectopically retained along the lateral ventricles. Genetic causes include FLNA abnormalities (classical X-linked PNH), rare variants in ARFGEF2, DCHS1, ERMARD, FAT4, INTS8, MAP1B, MCPH1, and NEDD4L, as well as several chromosomal abnormalities. We performed array-CGH in 106 patients with different malformations of cortical development (MCD) and looked for common pathways possibly involved in PNH. Forty-two patients, including two parent/proband couples, exhibited PNH associated or not with other brain abnormalities, 44 had polymicrogyria and 20 had rarer MCDs. We found an enrichment of either large rearrangements or cryptic copy number variants (CNVs) in PNH (15/42, 35.7%) vs polymicrogyria (4/44, 9.1%) (i.e., 5.6 times increased risk for PNH of carrying a pathogenic CNV). CNVs in seven genomic regions (2p11.2q12.1, 4p15, 14q11.2q12, 16p13.3, 19q13.33, 20q13.33, 22q11) represented novel, potentially causative, associations with PNH. Through in silico analysis of genes included in imbalances whose breakpoints were clearly detailed, we detected in 9/12 unrelated patients in our series and in 15/24 previously published patients, a significant (P
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- 2019
29. 6P Circulating tumour cells (CTCs) as biomarkers of resistance to the CDK4/6 inhibitor (CDK4/6i) palbociclib (P) in patients (pts) with ER+/HER2-negative advanced breast cancer (ABC)
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Laura Biganzoli, Chiara Biagioni, A. Di Leo, Valerio Conti, Amelia McCartney, F. De Luca, Luca Malorni, Giuseppe Curigliano, M. Benelli, I. Migliaccio, Alessandro Marco Minisini, Francesca Galardi, Emanuela Risi, Martina Bonechi, Dario Romagnoli, and Erica Moretti
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Oncology ,medicine.medical_specialty ,business.industry ,Advanced breast ,HER2 negative ,Cancer ,Hematology ,Palbociclib ,medicine.disease ,Internal medicine ,medicine ,In patient ,business - Published
- 2020
30. The landscape of epilepsy-related GATOR1 variants
- Author
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Johannes R. Lemke, Pia Zacher, Thomas Dorn, Laura Hernandez-Hernandez, Natasha E. Schoeler, Stéphanie Baulac, Sara Baldassari, Anne de Saint Martin, Eleni Panagiotakaki, Anne Fabienne Lepine, Markus Wolff, Arnaud Biraben, Renske Oegema, Edouard Hirsch, Anna Jansen, Charles Deckers, Nienke E. Verbeek, Fabienne Picard, Georg Dorfmüller, Sarah Ferrand-Sorbets, Barbora Benova, Francesca Bisulli, Inga Talvik, Kristin Lindstrom, Tilman Polster, Douglas R. Nordli, Tommaso Pippucci, Eva H. Brilstra, Shifteh Sattar, Erik H. Niks, Marie Line Jacquemont, Kees P.J. Braun, Karen Müller-Schlüter, Sanjay M. Sisodiya, Sarah Weckhuysen, Lysa Boissé Lomax, Sophie Julia, Brigitte Ricard-Mousnier, Mathilde Chipaux, Laura Licchetta, Gaetan Lesca, Bianca Berghuis, S. Krithika, Jamel Chelly, Renzo Guerrini, Hélène Catenoix, Annapurna Poduri, Melanie Jennesson, Pasquale Striano, Rikke S. Møller, Antonio Gambardella, Guillaume Achaz, Peter Uldall, Fabrice Bartolomei, Giuseppe d'Orsi, Laurence Faivre, Floor E. Jansen, An Sofie Schoonjans, Kevin Rostasy, Thomas Becher, Pavel Krsek, Julien Thevenon, Marjan J. A. van Kempen, Guido Rubboli, Cécile Marchal, Meral Balci, Boudewijn Gunning, Ilona Krey, Julitta de Bellescize, Veronique Darmency, Christopher J. Yuskaitis, Daniëlle de Jong, Giovanni Crichiutti, Paolo Tinuper, Katrien Stouffs, Valentin Sander, Anne-Sophie Lebre, Thomas Cloppenborg, Valerio Conti, Gabrielle Rudolf, Courtney Kiss, Eveline Hagebeuk, Caroline Nava, Eric LeGuern, Ilse Wegner, Christian Brandt, Martin Zenker, Simona Balestrini, Picard, Fabienne, Baldassari S., Picard F., Verbeek N.E., van Kempen M., Brilstra E.H., Lesca G., Conti V., Guerrini R., Bisulli F., Licchetta L., Pippucci T., Tinuper P., Hirsch E., de Saint Martin A., Chelly J., Rudolf G., Chipaux M., Ferrand-Sorbets S., Dorfmuller G., Sisodiya S., Balestrini S., Schoeler N., Hernandez-Hernandez L., Krithika S., Oegema R., Hagebeuk E., Gunning B., Deckers C., Berghuis B., Wegner I., Niks E., Jansen F.E., Braun K., de Jong D., Rubboli G., Talvik I., Sander V., Uldall P., Jacquemont M.-L., Nava C., Leguern E., Julia S., Gambardella A., d'Orsi G., Crichiutti G., Faivre L., Darmency V., Benova B., Krsek P., Biraben A., Lebre A.-S., Jennesson M., Sattar S., Marchal C., Nordli D.R., Lindstrom K., Striano P., Lomax L.B., Kiss C., Bartolomei F., Lepine A.F., Schoonjans A.-S., Stouffs K., Jansen A., Panagiotakaki E., Ricard-Mousnier B., Thevenon J., de Bellescize J., Catenoix H., Dorn T., Zenker M., Muller-Schluter K., Brandt C., Krey I., Polster T., Wolff M., Balci M., Rostasy K., Achaz G., Zacher P., Becher T., Cloppenborg T., Yuskaitis C.J., Weckhuysen S., Poduri A., Lemke J.R., Moller R.S., Baulac S., Institut du Cerveau et de la Moëlle Epinière = Brain and Spine Institute (ICM), Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Department of Genetics [Utrecht, the Netherlands], University Medical Center [Utrecht], Service de Génétique [HCL Groupement Hospitalier Est], Groupement Hospitalier Lyon-Est (GHE), Hospices Civils de Lyon (HCL)-Hospices Civils de Lyon (HCL), Centre de recherche en neurosciences de Lyon - Lyon Neuroscience Research Center (CRNL), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet [Saint-Étienne] (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Children's Hospital A. Meyer, Service de Neurologie [Strasbourg], CHU Strasbourg-Hopital Civil, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Department of Clinical and Experimental Epilepsy, University College of London [London] (UCL), Academic Center for Epileptology Kempenhaeghe & Maastricht UMC+ [Heeze], Danish Epilepsy Centre, Denmark and Aarhus University, Aarhus, Centre Hospitalier Universitaire de La Réunion (CHU La Réunion), Service de Génétique Cytogénétique et Embryologie [CHU Pitié-Salpêtrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Service de génétique médicale [Toulouse], CHU Toulouse [Toulouse], Centre de génétique - Centre de référence des maladies rares, anomalies du développement et syndromes malformatifs (CHU de Dijon), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), FHU TRANSLAD (CHU de Dijon), Université de Bourgogne (UB), Service de Neurophysiologie Clinique (CHU Dijon), CHU Pontchaillou [Rennes], Service de pédiatrie spécialisée et médecine infantile (neurologie, pneumologie, maladies héréditaires du métabolisme) [Hôpital de la Timone - APHM], Hôpital de la Timone [CHU - APHM] (TIMONE), Epilepsie, sommeil et explorations fonctionnelles neuropédiatriques, Hospices Civils de Lyon (HCL)-Hôpital Femme Mère Enfant, Equipe GAD (LNC - U1231), Lipides - Nutrition - Cancer [Dijon - U1231] (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement, Université Bourgogne Franche-Comté [COMUE] (UBFC), Département d'Epilepsie, Sommeil et Neurophysiologie Pédiatrique [HCL, Lyon], Hospices Civils de Lyon (HCL), Institute of Human Genetics, University Hospital Magdeburg, Institut de Systématique, Evolution, Biodiversité (ISYEB ), Muséum national d'Histoire naturelle (MNHN)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Groupement hospitalier Lyon-Est, Centre de recherche en neurosciences de Lyon (CRNL), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), and CHU Toulouse [Toulouse]-Hôpital Purpan [Toulouse]
- Subjects
Male ,0301 basic medicine ,Proband ,DEPDC5 ,SUDEP ,030105 genetics & heredity ,Bioinformatics ,Loss of Function Mutation/genetics ,Epilepsy ,INDEL Mutation ,Loss of Function Mutation ,mTORC1 pathway ,Genetics(clinical) ,Child ,Genetics (clinical) ,Multiprotein Complexes/genetics ,Brugada Syndrome ,DNA Copy Number Variation ,Brugada syndrome ,INDEL Mutation/genetics ,GTPase-Activating Proteins ,NPRL3 ,Seizure ,Phenotype ,Pedigree ,3. Good health ,Brugada Syndrome/genetics ,Child, Preschool ,Female ,Human ,Signal Transduction ,DNA Copy Number Variations ,Adolescent ,Seizures/complications ,Mechanistic Target of Rapamycin Complex 1/genetics ,DNA Copy Number Variations/genetics ,Mechanistic Target of Rapamycin Complex 1 ,Tumor Suppressor Proteins/genetics ,Article ,Focal cortical dysplasia ,03 medical and health sciences ,Seizures ,GTPase-Activating Proteins/genetics ,medicine ,Humans ,Genetic Predisposition to Disease ,Genetic focal epilepsy ,Epilepsy/complications ,Repressor Proteins/genetics ,business.industry ,GTPase-Activating Protein ,Tumor Suppressor Proteins ,Infant, Newborn ,Correction ,Infant ,Repressor Protein ,Cortical dysplasia ,medicine.disease ,ddc:616.8 ,Repressor Proteins ,030104 developmental biology ,Frontal lobe seizures ,[SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics ,Multiprotein Complexes ,Multiprotein Complexe ,Signal Transduction/genetics ,Human medicine ,business - Abstract
Purpose:\ud \ud To define the phenotypic and mutational spectrum of epilepsies related to DEPDC5, NPRL2 and NPRL3 genes encoding the GATOR1 complex, a negative regulator of the mTORC1 pathway.\ud \ud Methods:\ud \ud We analyzed clinical and genetic data of 73 novel probands (familial and sporadic) with epilepsy-related variants in GATOR1-encoding genes and proposed new guidelines for clinical interpretation of GATOR1 variants.\ud \ud Results:\ud \ud The GATOR1 seizure phenotype consisted mostly in focal seizures (e.g., hypermotor or frontal lobe seizures in 50%), with a mean age at onset of 4.4 years, often sleep-related and drug-resistant (54%), and associated with focal cortical dysplasia (20%). Infantile spasms were reported in 10% of the probands. Sudden unexpected death in epilepsy (SUDEP) occurred in 10% of the families. Novel classification framework of all 140 epilepsy-related GATOR1 variants (including the variants of this study) revealed that 68% are loss-of-function pathogenic, 14% are likely pathogenic, 15% are variants of uncertain significance and 3% are likely benign.\ud \ud Conclusion:\ud \ud Our data emphasize the increasingly important role of GATOR1 genes in the pathogenesis of focal epilepsies (>180 probands to date). The GATOR1 phenotypic spectrum ranges from sporadic early-onset epilepsies with cognitive impairment comorbidities to familial focal epilepsies, and SUDEP.
- Published
- 2018
31. 3d Human Brain Digital Histopatology
- Author
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Leonardo Sacconi, Ludovico Silvestri, Valerio Conti, M. Roffilli, Giacomo Mazzamuto, Mattia Neri, Irene Costantini, Renzo Guerrini, and Francesco S. Pavone
- Subjects
medicine.anatomical_structure ,Three dimensional imaging ,Cellular resolution ,Artificial neural network ,Cytoarchitecture ,business.industry ,Computer science ,medicine ,Pattern recognition ,Image processing ,Human brain ,Artificial intelligence ,business - Abstract
In this project we perform a quantitative analysis of brain cytoarchitecture to study the three-dimensional reconstruction of the human neural networks at cellular resolution expanding the histopathological studies to the third dimension.
- Published
- 2018
32. A Novel Strategy Combining Array-CGH, Whole-exome Sequencing and In Utero Electroporation in Rodents to Identify Causative Genes for Brain Malformations
- Author
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Carlos Cardoso, Antonio Falace, Renzo Guerrini, David Antony Keays, Alfonso Represa, Orsetta Zuffardi, Jenny C. Taylor, Usha Kini, Alistair T. Pagnamenta, Stefano Lise, Francesca Novara, Emmanuelle Buhler, Françoise Watrin, Agathe A. Deparis, Elena Parrini, Fabienne Schaller, Richard J. Leventer, Emilie Pallesi-Pocachard, Aurelie Carabalona, and Valerio Conti
- Subjects
General Immunology and Microbiology ,General Chemical Engineering ,General Neuroscience ,General Biochemistry, Genetics and Molecular Biology - Published
- 2017
33. Mutations of AKT3 are associated with a wide spectrum of developmental disorders including extreme megalencephaly
- Author
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Anna Lehman, Melinda Zombor, Mark O'Driscoll, Ute Moog, Natalia Gomez-Ospina, Valerio Conti, Adeline Jacquinet, Margot I. Van Allen, Sofia Ygberg, Andrew E. Timms, Renzo Guerrini, Jonathan A. Bernstein, Ghayda M. Mirzaa, Diana Alcantara, Fiona Stewart, Sarju G. Mehta, Oana Caluseriu, Sarah Collins, Ronit Mesterman, John M. Graham, Robert F. Hevner, Kaylee Park, William B. Dobyns, Enrico Alfei, László Sztriha, Gill Bejerano, Laura Baker, Anand Saggar, Chiara Pantaleoni, Aaron M. Wenger, Karen W. Gripp, Chi Cheng, and Harendra Guturu
- Subjects
0301 basic medicine ,Male ,Hemimegalencephaly ,Pathology ,medicine.medical_specialty ,RB155.5 ,Developmental Disabilities ,RB024 ,Biology ,Phosphatidylinositols ,Transfection ,akt3 ,RB127 ,03 medical and health sciences ,0302 clinical medicine ,mental disorders ,medicine ,Polymicrogyria ,Humans ,Immunoprecipitation ,Megalencephaly ,Kinase activity ,Child ,Genetic Association Studies ,Genetics ,RB151 ,Macrocephaly ,Brain ,RB057 ,Original Articles ,Cortical dysplasia ,medicine.disease ,Phenotype ,Magnetic Resonance Imaging ,3. Good health ,030104 developmental biology ,Heterotopia (medicine) ,HEK293 Cells ,Mutation ,Mutagenesis, Site-Directed ,Female ,Neurology (clinical) ,medicine.symptom ,Proto-Oncogene Proteins c-akt ,030217 neurology & neurosurgery - Abstract
Mutations of genes within the phosphatidylinositol-3-kinase (PI3K)-AKT-MTOR pathway are well known causes of brain overgrowth (megalencephaly) as well as segmental cortical dysplasia (such as hemimegalencephaly, focal cortical dysplasia and polymicrogyria). Mutations of the AKT3 gene have been reported in a few individuals with brain malformations, to date. Therefore, our understanding regarding the clinical and molecular spectrum associated with mutations of this critical gene is limited, with no clear genotype-phenotype correlations. We sought to further delineate this spectrum, study levels of mosaicism and identify genotype-phenotype correlations of AKT3-related disorders. We performed targeted sequencing of AKT3 on individuals with these phenotypes by molecular inversion probes and/or Sanger sequencing to determine the type and level of mosaicism of mutations. We analysed all clinical and brain imaging data of mutation-positive individuals including neuropathological analysis in one instance. We performed ex vivo kinase assays on AKT3 engineered with the patient mutations and examined the phospholipid binding profile of pleckstrin homology domain localizing mutations. We identified 14 new individuals with AKT3 mutations with several phenotypes dependent on the type of mutation and level of mosaicism. Our comprehensive clinical characterization, and review of all previously published patients, broadly segregates individuals with AKT3 mutations into two groups: patients with highly asymmetric cortical dysplasia caused by the common p.E17K mutation, and patients with constitutional AKT3 mutations exhibiting more variable phenotypes including bilateral cortical malformations, polymicrogyria, periventricular nodular heterotopia and diffuse megalencephaly without cortical dysplasia. All mutations increased kinase activity, and pleckstrin homology domain mutants exhibited enhanced phospholipid binding. Overall, our study shows that activating mutations of the critical AKT3 gene are associated with a wide spectrum of brain involvement ranging from focal or segmental brain malformations (such as hemimegalencephaly and polymicrogyria) predominantly due to mosaic AKT3 mutations, to diffuse bilateral cortical malformations, megalencephaly and heterotopia due to constitutional AKT3 mutations. We also provide the first detailed neuropathological examination of a child with extreme megalencephaly due to a constitutional AKT3 mutation. This child has one of the largest documented paediatric brain sizes, to our knowledge. Finally, our data show that constitutional AKT3 mutations are associated with megalencephaly, with or without autism, similar to PTEN-related disorders. Recognition of this broad clinical and molecular spectrum of AKT3 mutations is important for providing early diagnosis and appropriate management of affected individuals, and will facilitate targeted design of future human clinical trials using PI3K-AKT pathway inhibitors.
- Published
- 2017
34. De novo mutations of the ATP6V1A gene cause developmental encephalopathy with epilepsy
- Author
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Chihiro Ohba, Akgun Olmez Turker, Burcu Albuz, Fabio Benfenati, Kazuhiro Ogata, Naomichi Matsumoto, Ashley L. Siniard, Keri Ramsey, Alessandro Esposito, Davide Mei, Carla Marini, Mitsuhiro Kato, Chris Balak, C Nur Semerci Gündüz, Renzo Guerrini, Vinodh Narayanan, Keiko Yanagihara, Elisa Belmonte, Mitsuko Nakashima, Valerio Conti, Masaaki Shiina, Anna Fassio, Nobuhiko Okamoto, Luca Maragliano, and Hirotomo Saitsu
- Subjects
Male ,hippocampus ,Endocytic cycle ,autophagosome ,cell organelle ,Vesicular Transport Proteins ,whole exome sequencing ,stereospecificity ,developmental epileptic encephalopathy ,Proton transport ,v type proton transporting adenosine triphosphatase ,genetics ,nuclear magnetic resonance imaging ,Child ,Cells, Cultured ,Regulation of gene expression ,Neurons ,Mutation ,clinical article ,Adolescent ,Animals ,Brain/diagnostic imaging ,Brain Diseases/complications/*genetics/pathology ,Cohort Studies ,Epilepsy/complications/*genetics/pathology ,Female ,Gene Expression Regulation/genetics ,HEK293 Cells ,Humans ,Lysosomal-Associated Membrane Protein 1/metabolism ,Lysosomes/metabolism/pathology ,Models, Molecular ,Mutation/*genetics ,Neurons/metabolism/pathology/ultrastructure ,Rats ,Synapses/metabolism/pathology ,Vacuolar Proton-Translocating ATPases/*ge ,Brain ,gene expression regulation ,cohort analysis ,Cell biology ,developmental delay ,priority journal ,protein stability ,sequence alignment ,brain nerve cell ,nerve cell ,ATP6V1A protein, human ,mutational analysis ,Vacuolar Proton-Translocating ATPases ,EEG abnormality ,neurite outgrowth ,phenotype ,diagnostic imaging ,embryo ,complication ,Article ,animal tissue ,febrile convulsion ,03 medical and health sciences ,early endosome antigen 1 ,lysosomes ,Exome Sequencing ,V-ATPase ,case report ,human ,protein expression ,Loss function ,developmental encephalopathy ,Epilepsy ,catalysis ,human cell ,proton transporting adenosine triphosphatase ,functional connectivity ,school child ,030104 developmental biology ,proton transporting adenosine triphosphate synthase ,Synapses ,Neurology (clinical) ,proton transport ,molecular model ,0301 basic medicine ,medicine.disease_cause ,fluorescence microscopy ,synapse ,protein folding ,homeostasis ,rat ,animal ,neurite elongation ,brain disease ,Brain Diseases ,Chemistry ,intractable epilepsy ,ultrastructure ,unclassified drug ,female ,medicine.anatomical_structure ,v-ATPase ,HEK293 cell line ,lysosome ,quadriplegia ,ATP6V1A gene ,amino acid substitution ,Neurite ,HEK293T cell line ,Lysosomal-Associated Membrane Protein 1 ,Lysosome ,medicine ,controlled study ,gene ,developmental epileptic encephalopathy, lysosomes, neurite elongation, synapse, v-ATPase, Neurology (clinical) ,lymphoblast ,cell culture ,model ,nonhuman ,missense mutation ,vesicular transport protein ,Original Articles ,heterozygote ,clinical feature ,Gene Expression Regulation ,lysosome associated membrane protein 1 ,gene expression ,pathology ,metabolism - Abstract
Using whole exome sequencing, Fassio et al. identify de novo mutations in ATP6V1A, encoding the A subunit of v-ATPase, in four patients with developmental encephalopathies and epilepsy. Functional and expression studies demonstrate impaired lysosomal homeostasis, defective neurite elongation and loss of excitatory inputs in cultured neurons., V-type proton (H+) ATPase (v-ATPase) is a multi-subunit proton pump that regulates pH homeostasis in all eukaryotic cells; in neurons, v-ATPase plays additional and unique roles in synapse function. Through whole exome sequencing, we identified de novo heterozygous mutations (p.Pro27Arg, p.Asp100Tyr, p.Asp349Asn, p.Asp371Gly) in ATP6V1A, encoding the A subunit of v-ATPase, in four patients with developmental encephalopathy with epilepsy. Early manifestations, observed in all patients, were developmental delay and febrile seizures, evolving to encephalopathy with profound delay, hypotonic/dyskinetic quadriparesis and intractable multiple seizure types in two patients (p.Pro27Arg, p.Asp100Tyr), and to moderate delay with milder epilepsy in the other two (p.Asp349Asn, p.Asp371Gly). Modelling performed on the available prokaryotic and eukaryotic structures of v-ATPase predicted p.Pro27Arg to perturb subunit interaction, p.Asp100Tyr to cause steric hindrance and destabilize protein folding, p.Asp349Asn to affect the catalytic function and p.Asp371Gly to impair the rotation process, necessary for proton transport. We addressed the impact of p.Asp349Asn and p.Asp100Tyr mutations on ATP6V1A expression and function by analysing ATP6V1A-overexpressing HEK293T cells and patients’ lymphoblasts. The p.Asp100Tyr mutant was characterized by reduced expression due to increased degradation. Conversely, no decrease in expression and clearance was observed for p.Asp349Asn. In HEK293T cells overexpressing either pathogenic or control variants, p.Asp349Asn significantly increased LysoTracker® fluorescence with no effects on EEA1 and LAMP1 expression. Conversely, p.Asp100Tyr decreased both LysoTracker® fluorescence and LAMP1 levels, leaving EEA1 expression unaffected. Both mutations decreased v-ATPase recruitment to autophagosomes, with no major impact on autophagy. Experiments performed on patients’ lymphoblasts using the LysoSensor™ probe revealed lower pH of endocytic organelles for p.Asp349Asn and a reduced expression of LAMP1 with no effect on the pH for p.Asp100Tyr. These data demonstrate gain of function for p.Asp349Asn characterized by an increased proton pumping in intracellular organelles, and loss of function for p.Asp100Tyr with decreased expression of ATP6V1A and reduced levels of lysosomal markers. We expressed p.Asp349Asn and p.Asp100Tyr in rat hippocampal neurons and confirmed significant and opposite effects in lysosomal labelling. However, both mutations caused a similar defect in neurite elongation accompanied by loss of excitatory inputs, revealing that altered lysosomal homeostasis markedly affects neurite development and synaptic connectivity. This study provides evidence that de novo heterozygous ATP6V1A mutations cause a developmental encephalopathy with a pathomechanism that involves perturbations of lysosomal homeostasis and neuronal connectivity, uncovering a novel role for v-ATPase in neuronal development.
- Published
- 2017
35. Multimodal fiber-probe spectroscopy as a clinical tool for diagnosing and classifying biological tissues
- Author
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Renzo Guerrini, Francesco S. Pavone, Riccardo Cicchi, Suresh Anand, Gabriella Nesi, Anna Maria Buccoliero, Mauro Gacci, Flavio Giordano, Marco Carini, R. Fantechi, and Valerio Conti
- Subjects
Optical fiber ,Materials science ,Diffuse reflectance infrared fourier transform ,Laser diode ,business.industry ,Near-infrared spectroscopy ,Fluorescence spectroscopy ,law.invention ,symbols.namesake ,Halogen lamp ,law ,symbols ,Optoelectronics ,Raman spectroscopy ,business ,Spectroscopy - Abstract
An optical fiber probe for multimodal spectroscopy was designed, developed and used for tissue diagnostics. The probe, based on a fiber bundle with optical fibers of various size and properties, allows performing spectroscopic measurements with different techniques, including fluorescence, Raman, and diffuse reflectance, using the same probe. Two visible laser diodes were used for fluorescence spectroscopy, a laser diode emitting in the NIR was used for Raman spectroscopy, and a fiber-coupled halogen lamp for diffuse reflectance. The developed probe was successfully employed for diagnostic purposes on various tissues, including brain and bladder. In particular, the device allowed discriminating healthy tissue from both tumor and dysplastic tissue as well as to perform tumor grading. The diagnostic capabilities of the method, determined using a cross-validation method with a leave-one-out approach, demonstrated high sensitivity and specificity for all the examined samples, as well as a good agreement with histopathological examination performed on the same samples. The obtained results demonstrated that the multimodal approach is crucial for improving diagnostic capabilities with respect to what can be obtained from individual techniques. The experimental setup presented here can improve diagnostic capabilities on a broad range of tissues and has the potential of being used clinically for guiding surgical resection in the near future.
- Published
- 2017
36. Genetic Basis of Brain Malformations
- Author
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William B. Dobyns, Valerio Conti, Renzo Guerrini, and Elena Parrini
- Subjects
0301 basic medicine ,Hemimegalencephaly ,Pathology ,medicine.medical_specialty ,Microcephaly ,Genetic heterogeneity ,Lissencephaly ,Review Article ,Cortical dysplasia ,Biology ,medicine.disease ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,nervous system ,Genetics ,medicine ,Polymicrogyria ,Megalencephaly ,Cerebellar hypoplasia ,030217 neurology & neurosurgery ,Genetics (clinical) - Abstract
Malformations of cortical development (MCD) represent a major cause of developmental disabilities, severe epilepsy, and reproductive disadvantage. Genes that have been associated to MCD are mainly involved in cell proliferation and specification, neuronal migration, and late cortical organization. Lissencephaly-pachygyria-severe band heterotopia are diffuse neuronal migration disorders causing severe global neurological impairment. Abnormalities of the LIS1, DCX, ARX, RELN, VLDLR, ACTB, ACTG1, TUBG1, KIF5C, KIF2A, and CDK5 genes have been associated with these malformations. More recent studies have also established a relationship between lissencephaly, with or without associated microcephaly, corpus callosum dysgenesis as well as cerebellar hypoplasia, and at times, a morphological pattern consistent with polymicrogyria with mutations of several genes (TUBA1A, TUBA8, TUBB, TUBB2B, TUBB3, and DYNC1H1), regulating the synthesis and function of microtubule and centrosome key components and hence defined as tubulinopathies. MCD only affecting subsets of neurons, such as mild subcortical band heterotopia and periventricular heterotopia, have been associated with abnormalities of the DCX, FLN1A, and ARFGEF2 genes and cause neurological and cognitive impairment that vary from severe to mild deficits. Polymicrogyria results from abnormal late cortical organization and is inconstantly associated with abnormal neuronal migration. Localized polymicrogyria has been associated with anatomo-specific deficits, including disorders of language and higher cognition. Polymicrogyria is genetically heterogeneous, and only in a small minority of patients, a definite genetic cause has been identified. Megalencephaly with normal cortex or polymicrogyria by MRI imaging, hemimegalencephaly and focal cortical dysplasia can all result from mutations in genes of the PI3K-AKT-mTOR pathway. Postzygotic mutations have been described for most MCD and can be limited to the dysplastic tissue in the less diffuse forms.
- Published
- 2017
37. Fiber-probe optical spectroscopy discriminates normal brain from focal cortical dysplasia in pediatric subjects
- Author
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Renzo Guerrini, Anna Maria Buccoliero, Valerio Conti, Riccardo Cicchi, Francesco S. Pavone, Suresh Anand, and Flavio Giordano
- Subjects
Excitation wavelength ,Materials science ,medicine.diagnostic_test ,business.industry ,05 social sciences ,Normal tissue ,050801 communication & media studies ,Magnetic resonance imaging ,Cortical dysplasia ,medicine.disease ,01 natural sciences ,010309 optics ,symbols.namesake ,0508 media and communications ,Optics ,Nuclear magnetic resonance ,0103 physical sciences ,medicine ,symbols ,Fiber probe ,A fibers ,business ,Spectroscopy ,Raman spectroscopy - Abstract
Focal cortical dysplasia (FCD) is an abnormality in the cerebral cortex that is caused by malformations during cortical development. Currently, magnetic resonance imaging (MRI) and electro-corticography (ECoG) are used for detecting FCD. On the downside, MRI is very much insensitive to small malformations in the brain, while ECoG is an invasive and time consuming procedure. Recently, optical techniques were widely exploited as a minimally invasive and quantitative approaches for disease diagnosis. These techniques include fluorescence and Raman spectroscopy. The aim of this investigation is to study the diagnostic performances of optical spectroscopy incorporating fluorescence (at 378 nm and 445 nm excitation wavelengths) and Raman spectroscopy (at 785 nm excitation) for the discrimination of FCD from normal brain in pediatric subjects. The study included 10 normal and 17 FCD tissue sites from 3 normal and 7 FCD samples. The emission spectra of FCD at 378 nm excitation wavelength presented a blue-shifted peak with respect to normal tissue. Prominent spectral differences between normal and FCD tissue were observed at 1298 cm -1 , 1302 cm -1 , 1445 cm -1 and 1660 cm -1 using Raman spectroscopy. Tissue classification models were developed using a multivariate statistical method, principal component analysis. This study demonstrates that a combined spectroscopic approach can provide a better diagnostic capability for classifying normal and FCD tissues. Further, the implementation of the technology within a fiber probe could open the way for in vivo diagnostics and intra-operative surgical guidance.
- Published
- 2017
38. Human Mutations Associated With Brain Malformations Resulting in Hyperexcitability in Rodents
- Author
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Renzo Guerrini and Valerio Conti
- Subjects
Childhood epilepsy ,Epilepsy ,medicine.anatomical_structure ,Autism spectrum disorder ,Cerebral cortex ,Neuronal migration ,medicine ,medicine.disease ,Psychology ,Phenotype ,Neuroscience ,Motor disability - Abstract
The development of the human cerebral cortex is a complex and tightly organized process. Disruption of any of the overlapping steps that contribute to this process can result in a wide range of developmental brain disorders (DBD) that are associated, in various combinations, with severe childhood epilepsy, intellectual and motor disability, and autism spectrum disorder. Pediatric epilepsies are among the most devastating neurologic disorders, and epilepsy research has a long history of comparative anatomical and physiological studies on a range of mammalian species. A number of animal models have been generated to help researchers in investigating basic mechanisms underlying DBD, and implementing new diagnostic approaches, as well as antiepileptic medications. Here, we review to what extent different rodent models have helped researchers elucidating how mutations in specific genes cause brain malformations and epilepsy, as well as how specific treatments may help in alleviating the phenotype.
- Published
- 2017
39. Focal dysplasia of the cerebral cortex and infantile spasms associated with somatic 1q21.1-q44 duplication including theAKT3gene
- Author
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Seung Tae Baek, Gianna Baroni, Valerio Conti, Renzo Guerrini, Flavio Giordano, Carmen Barba, Sabrina Giglio, Anna Maria Buccoliero, Davide Mei, Marilena Pantaleo, and Joseph G. Gleeson
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Hemimegalencephaly ,Anatomy ,Biology ,Cortical dysplasia ,medicine.disease ,White matter ,Epilepsy ,medicine.anatomical_structure ,Cerebral cortex ,Cortex (anatomy) ,Chromosomal region ,Genetics ,medicine ,Megalencephaly ,Genetics (clinical) - Abstract
Somatic and germline duplications or activating mutations of AKT3 have been reported in patients with hemimegalencephaly and megalencephaly. We performed array comparative genomic hybridization on brain tissue and blood in 16 consecutive patients with symptomatic epilepsy due to focal or multilobar malformations of cortical development who underwent surgical treatment of epilepsy. One patient with infantile spasms and a dysplastic left frontal lobe harboured a somatic trisomy of the 1q21.1-q44 chromosomal region, encompassing the AKT3 gene, in the dysplastic brain tissue but not in blood and saliva. Histopathology revealed severe cortical dyslamination, a thin cortex in the premotor area with microgyri and microsulci, immature neurons with disoriented dendrites and areas of cortical heterotopia in the sub-cortical white matter. These cytoarchitectural changes are close to those defining type Ib focal cortical dysplasia. Immunohistochemistry in brain specimens showed hyperactivation of the PI3K/AKT/mTOR pathway. These findings indicate that AKT3 upregulation may cause focal malformations of cortical development. There appears to be an etiologic continuum between hemimegalencephaly and focal cortical dysplastic lesions. The extent of brain malformations due to AKT3 upregulation may be related to the embryonic stage when the post-zygotic gene alteration occurs.
- Published
- 2014
40. PIK3CA-associated developmental disorders exhibit distinct classes of mutations with variable expression and tissue distribution
- Author
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Lim Jiin Yin, Beth Martin, Mark J. Stephan, Mariana Aracena, Cynthia J. Curry, Inge Krägeloh-Mann, Karen W. Gripp, Koenraad Devriendt, Michael Painter, Livia Garavelli, William B. Dobyns, Rachel Straussberg, Agustina Lanoel, Marie-Claude Addor, Margaret L. McKinnon, Luigi Boccuto, John Graham, Katrina Tatton-Brown, James D. Reggin, Jay Shendure, Colin C. Pritchard, Charles E. Schwartz, Mary Ella M Pierpont, Ian A. Glass, Fiona Stewart, Sulagna C. Saitta, Angeline Hwei Meeng Lai, Evan A. Boyle, Erin Torti, Anne Goriely, Michael T. Gabbett, Melanie Napier, Nicole Martin, Melissa T. Carter, Lisa Worgan, Renzo Guerrini, Katta M. Girisha, Ghayda M. Mirzaa, Chitra Prasad, Rachael Bradshaw, Leah W. Burke, Martin Kircher, Hulya Kayserilli, Andrew E. Timms, Jane Juusola, Karen D. Tsuchiya, Catherine E. Keegan, Robert L. Conway, David Chitayat, Kaylee Park, Hilde Van Esch, Aditi Shah Parikh, Maria R. Cordisco, Valerio Conti, Sondhya Ghedia, Raoul C.M. Hennekam, Sarah Collins, Bridget C. O’Connor, Stephen R. Braddock, Carissa Olds, ANS - Complex Trait Genetics, APH - Amsterdam Public Health, and Paediatric Genetics
- Subjects
0301 basic medicine ,Genetics ,Sanger sequencing ,Somatic cell ,Class I Phosphatidylinositol 3-Kinases/genetics ,Female ,Genetic Association Studies ,High-Throughput Nucleotide Sequencing ,Humans ,Infant ,Male ,Malformations of Cortical Development/genetics ,Mosaicism ,Mutation ,Phenotype ,Tissue Distribution ,Vascular Malformations/genetics ,General Medicine ,Biology ,Molecular biology ,3. Good health ,Variable Expression ,03 medical and health sciences ,symbols.namesake ,030104 developmental biology ,Targeted ngs ,symbols ,Amplicon sequencing ,Tissue distribution ,Class I Phosphatidylinositol 3-Kinases ,neoplasms ,Research Article - Abstract
Mosaicism is increasingly recognized as a cause of developmental disorders with the advent of next-generation sequencing (NGS). Mosaic mutations of PIK3CA have been associated with the widest spectrum of phenotypes associated with overgrowth and vascular malformations. We performed targeted NGS using 2 independent deep-coverage methods that utilize molecular inversion probes and amplicon sequencing in a cohort of 241 samples from 181 individuals with brain and/or body overgrowth. We identified PIK3CA mutations in 60 individuals. Several other individuals (n = 12) were identified separately to have mutations in PIK3CA by clinical targeted-panel testing (n = 6), whole-exome sequencing (n = 5), or Sanger sequencing (n = 1). Based on the clinical and molecular features, this cohort segregated into three distinct groups: (a) severe focal overgrowth due to low-level but highly activating (hotspot) mutations, (b) predominantly brain overgrowth and less severe somatic overgrowth due to less-activating mutations, and (c) intermediate phenotypes (capillary malformations with overgrowth) with intermediately activating mutations. Sixteen of 29 PIK3CA mutations were novel. We also identified constitutional PIK3CA mutations in 10 patients. Our molecular data, combined with review of the literature, show that PIK3CA-related overgrowth disorders comprise a discontinuous spectrum of disorders that correlate with the severity and distribution of mutations., The clinical and molecular spectrum of PIK3CA-related developmental disorders are correlated with types of mutations, tissue distributions, and levels of mosaicism with the clinical phenotype.
- Published
- 2016
41. Association of MTOR Mutations With Developmental Brain Disorders, Including Megalencephaly, Focal Cortical Dysplasia, and Pigmentary Mosaicism
- Author
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Carleton Goold, Nadia Solovieff, William B. Dobyns, Richard A. Gibbs, Elisa Rahikkala, Jonathan D. Biag, Colleen F. Macmurdo, Eric Boerwinkle, Sonya A. Gunter, Sandra L. Poliachik, Brian H.Y. Chung, Christine D. Wilson, Evan A. Boyle, Scott Mahan, Robert F. Hevner, Russell P. Saneto, Katta M. Girisha, Rebecca Leary, Wendy Winckler, Molly Weaver, Sharon S. McDaniel, Jay Shendure, Laura A. Jansen, Shanming Liu, Shalini N. Jhangiani, Andrew E. Timms, William R. Sellers, Gisele Ishak, Michael Morrissey, Michael O. Dorschner, Donna M. Muzny, Catarina D. Campbell, Jeffrey G. Ojemann, Beth Martin, Edward J. Novotny, Renzo Guerrini, Ghayda M. Mirzaa, Leon Murphy, Valerio Conti, Jonathan A. Bernstein, James R. Lupski, Sarah Collins, Suchithra Menon, Carissa Olds, and Kit San Yeung
- Subjects
0301 basic medicine ,Adult ,Male ,Hemimegalencephaly ,Pathology ,medicine.medical_specialty ,Adolescent ,Developmental Disabilities ,Neurogenetics ,Nerve Tissue Proteins ,Biology ,Mechanistic Target of Rapamycin Complex 1 ,medicine.disease_cause ,03 medical and health sciences ,Epilepsy ,Young Adult ,0302 clinical medicine ,medicine ,Animals ,Humans ,Megalencephaly ,Amino Acids ,Child ,Exome sequencing ,Cells, Cultured ,Genetic Association Studies ,Retrospective Studies ,Cerebral Cortex ,Neurons ,Mutation ,Mosaicism ,TOR Serine-Threonine Kinases ,Macrocephaly ,Cortical dysplasia ,medicine.disease ,Embryo, Mammalian ,Rats ,Malformations of Cortical Development ,030104 developmental biology ,Gene Expression Regulation ,Child, Preschool ,Multiprotein Complexes ,Intercellular Signaling Peptides and Proteins ,Female ,Neurology (clinical) ,medicine.symptom ,030217 neurology & neurosurgery - Abstract
Importance Focal cortical dysplasia (FCD), hemimegalencephaly, and megalencephaly constitute a spectrum of malformations of cortical development with shared neuropathologic features. These disorders are associated with significant childhood morbidity and mortality. Objective To identify the underlying molecular cause of FCD, hemimegalencephaly, and diffuse megalencephaly. Design, Setting, and Participants Patients with FCD, hemimegalencephaly, or megalencephaly (mean age, 11.7 years; range, 2-32 years) were recruited from Pediatric Hospital A. Meyer, the University of Hong Kong, and Seattle Children’s Research Institute from June 2012 to June 2014. Whole-exome sequencing (WES) was performed on 8 children with FCD or hemimegalencephaly using standard-depth (50-60X) sequencing in peripheral samples (blood, saliva, or skin) from the affected child and their parents and deep (150-180X) sequencing in affected brain tissue. Targeted sequencing and WES were used to screen 93 children with molecularly unexplained diffuse or focal brain overgrowth. Histopathologic and functional assays of phosphatidylinositol 3-kinase–AKT (serine/threonine kinase)–mammalian target of rapamycin (mTOR) pathway activity in resected brain tissue and cultured neurons were performed to validate mutations. Main Outcomes and Measures Whole-exome sequencing and targeted sequencing identified variants associated with this spectrum of developmental brain disorders. Results Low-level mosaic mutations ofMTORwere identified in brain tissue in 4 children with FCD type 2a with alternative allele fractions ranging from 0.012 to 0.086. Intermediate-level mosaic mutation ofMTOR(p.Thr1977Ile) was also identified in 3 unrelated children with diffuse megalencephaly and pigmentary mosaicism in skin. Finally, a constitutional de novo mutation ofMTOR(p.Glu1799Lys) was identified in 3 unrelated children with diffuse megalencephaly and intellectual disability. Molecular and functional analysis in 2 children with FCD2a from whom multiple affected brain tissue samples were available revealed a mutation gradient with an epicenter in the most epileptogenic area. When expressed in cultured neurons, allMTORmutations identified here drive constitutive activation of mTOR complex 1 and enlarged neuronal size. Conclusions and Relevance In this study, mutations ofMTORwere associated with a spectrum of brain overgrowth phenotypes extending from FCD type 2a to diffuse megalencephaly, distinguished by different mutations and levels of mosaicism. These mutations may be sufficient to cause cellular hypertrophy in cultured neurons and may provide a demonstration of the pattern of mosaicism in brain and substantiate the link between mosaic mutations ofMTORand pigmentary mosaicism in skin.
- Published
- 2016
42. Towards automated neuron tracing via global and local 3D image analysis
- Author
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Paolo Soda, Ludovica Acciai, Renzo Guerrini, Francesco S. Pavone, Giulio Iannello, Valerio Conti, and Irene Costantini
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0301 basic medicine ,Ground truth ,Computational neuroscience ,Artificial neural network ,business.industry ,Computer science ,Phase (waves) ,Human brain ,Tracing ,Sample (graphics) ,03 medical and health sciences ,030104 developmental biology ,medicine.anatomical_structure ,Two-photon excitation microscopy ,medicine ,Computer vision ,Noise (video) ,Artificial intelligence ,business ,TRACE (psycholinguistics) - Abstract
The reconstruction of the neural network is essential in computational neuroscience. Here, we present an automatic algorithm to trace single neuron projections based on two core algorithmic ideas: a global step segmenting all neuron bodies and their projections and a local growing phase that accommodates to the nonuniform illumination and to the noise of the sample. We tested our algorithm on two 3D stacks of two-photon images acquired from a human dysplastic brain sample. The results show that the traces produced are statistically equivalent to the ground truth, according to the Friedman and Li tests. Furthermore, we found that our algorithm outperforms other state-of-the-art methods.
- Published
- 2016
43. Probing focal cortical dysplasia in formalin fixed samples using tissue optical spectroscopy
- Author
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Renzo Guerrini, Anna Maria Buccoliero, Francesco S. Pavone, Valerio Conti, Riccardo Cicchi, Flavio Giordano, and Suresh Anand
- Subjects
Materials science ,medicine.diagnostic_test ,Computed tomography ,Magnetic resonance imaging ,Context (language use) ,Formalin fixed ,Cortical dysplasia ,medicine.disease ,01 natural sciences ,Intensity (physics) ,010309 optics ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,Nuclear magnetic resonance ,Cerebral cortex ,0103 physical sciences ,medicine ,Spectroscopy ,030217 neurology & neurosurgery - Abstract
Focal cortical dysplasia (FCD) is one of most common causes of intractable epilepsy in pediatric population and these are often insensitive to anti-epileptic drugs. FCD is characterized by a disarray in localized regions of the cerebral cortex and abnormal neurons which results them to misfire with incorrect signals. Resective neurosurgery to remove or disconnect the affected parts from the rest of the brain seems to be a viable option to treat FCD. Before neurosurgery the subject could undergo imaging studies including magnetic resonance imaging (MRI) or computed tomography (CT) scans. On the downside FCD could be elusive in MRI images and may be practically invisible in CT scans. Furthermore, unnecessary removal of normal tissues is to be taken into consideration as this could lead to neurological defects. In this context, optical spectroscopy have been widely investigated as an alternative technique for the detection of abnormal tissues in different organ sites. Disease progression is accompanied by a number of architectural, biochemical and morphological changes. These variations are reflected in the spectral intensity and line shape. Here, in this proof of concept study we propose to investigate the application of tissue optical spectroscopy based on fluorescence excitation at two wavelength 378 and 445 nm coupled along with Raman spectroscopy for the detection of FCD on formalin fixed tissue specimens from pediatric subjects. For fluorescence at both the excitation wavelengths FCD showed a decreased intensity at longer wavelength when compared to normal tissues. Also, differences exist in the Raman spectral profiles of normal and FCD.
- Published
- 2016
44. Germline and somatic mutations in the MTOR gene in focal cortical dysplasia and epilepsy
- Author
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Susan M. Hiatt, Sarah Weckhuysen, E. Martina Bebin, Stéphanie Baulac, Virginie Lambrecq, Sarah Ferrand-Sorbets, Line H.G. Larsen, Valerio Conti, Georg Dorfmüller, Jeremy W. Prokop, Elise Marsan, Rikke S. Møller, Gregory M. Cooper, Kevin M. Bowling, Eric LeGuern, Davide Mei, Pierre de la Grange, Mathilde Chipaux, Valérie Taly, Renzo Guerrini, Guido Rubboli, The Danish Epilepsy Centre Filadelfia [Dianalund, Denmark], Institute for Regional Health Services [Odense, Denmark], University of Southern Denmark (SDU), Centre de référence des épilepsies rares [CHU Pitié-Salpêtrière], Unité fonctionnelle d'épilepsie [CHU Pitié-Salpêtrière], Service de Neurologie [CHU Pitié-Salpêtrière], IFR70-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-IFR70-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Service de Neurologie [CHU Pitié-Salpêtrière], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Institut du Cerveau et de la Moëlle Epinière = Brain and Spine Institute (ICM), Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Service de neurochirurgie pédiatrique [Fondation Rothschild, Paris], Fondation Rothschild, Médecine Personnalisée, Pharmacogénomique, Optimisation Thérapeutique (MEPPOT - U1147), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), HudsonAlpha Institute for Biotechnology [Huntsville, AL], CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Pediatric Neurology & Neurogenetics Unit and Laboratories, Università degli Studi di Firenze = University of Florence [Firenze] (UNIFI)-Children's Hospital A. Meyer, TALY, Valerie, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-IFR70-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Service de Neurologie [CHU Pitié-Salpêtrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Institut du Cerveau = Paris Brain Institute (ICM), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), and Università degli Studi di Firenze = University of Florence (UniFI)-Children's Hospital A. Meyer
- Subjects
0301 basic medicine ,[SDV]Life Sciences [q-bio] ,Autosomal dominant nocturnal frontal lobe epilepsy ,Germline ,Article ,03 medical and health sciences ,Epilepsy ,0302 clinical medicine ,Germline mutation ,medicine ,Journal Article ,Missense mutation ,[SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] ,Genetics (clinical) ,Genetics ,business.industry ,Pharmacology. Therapy ,Macrocephaly ,Cortical dysplasia ,medicine.disease ,3. Good health ,[SDV] Life Sciences [q-bio] ,030104 developmental biology ,Epilepsy syndromes ,Cancer research ,[SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] ,Neurology (clinical) ,Human medicine ,medicine.symptom ,business ,030217 neurology & neurosurgery - Abstract
OBJECTIVE: To assess the prevalence of somatic MTOR mutations in focal cortical dysplasia (FCD) and of germline MTOR mutations in a broad range of epilepsies.METHODS: We collected 20 blood-brain paired samples from patients with FCD and searched for somatic variants using deep-targeted gene panel sequencing. Germline mutations in MTOR were assessed in a French research cohort of 93 probands with focal epilepsies and in a diagnostic Danish cohort of 245 patients with a broad range of epilepsies. Data sharing among collaborators allowed us to ascertain additional germline variants in MTOR.RESULTS: We detected recurrent somatic variants (p.Ser2215Phe, p.Ser2215Tyr, and p.Leu1460Pro) in the MTOR gene in 37% of participants with FCD II and showed histologic evidence for activation of the mTORC1 signaling cascade in brain tissue. We further identified 5 novel de novo germline missense MTOR variants in 6 individuals with a variable phenotype from focal, and less frequently generalized, epilepsies without brain malformations, to macrocephaly, with or without moderate intellectual disability. In addition, an inherited variant was found in a mother-daughter pair with nonlesional autosomal dominant nocturnal frontal lobe epilepsy.CONCLUSIONS: Our data illustrate the increasingly important role of somatic mutations of the MTOR gene in FCD and germline mutations in the pathogenesis of focal epilepsy syndromes with and without brain malformation or macrocephaly.
- Published
- 2016
45. Characterisation of mutations of the phosphoinositide-3-kinase regulatory subunit, PIK3R2, in perisylvian polymicrogyria: a next-generation sequencing study
- Author
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Melissa T. Carter, Ed Blair, Evan A. Boyle, Yoko Narumi-Kishimoto, Sarah Ahmed, Lorenzo Tattini, Sarah Collins, Davide Mei, Patrick Nitschke, Jamel Chelly, Valerio Conti, Amy Goldstein, Renzo Guerrini, Ghayda M. Mirzaa, Allison L. Goetsch, Kathreen Johnston, Jay Shendure, Catharine Harris, Carissa Olds, Christopher Barnett, Kathryn Friend, Andrew E. Timms, Beth Martin, Anne Slavotinek, Jean-François Deleuze, Elena Parrini, William B. Dobyns, Christopher D. Smyser, Robert B. Hufnagel, and Sarah Barnett
- Subjects
Adolescent ,Population ,DNA Mutational Analysis ,Biology ,Bioinformatics ,medicine.disease_cause ,DNA sequencing ,Article ,MPPH syndrome ,03 medical and health sciences ,Young Adult ,Phosphatidylinositol 3-Kinases ,0302 clinical medicine ,Intellectual Disability ,Polymicrogyria ,medicine ,megalencephaly ,Missense mutation ,Humans ,Abnormalities, Multiple ,Congenital bilateral perisylvian syndrome ,polymicrogyria ,education ,Child ,030304 developmental biology ,Genetic testing ,Genetics ,0303 health sciences ,education.field_of_study ,Mutation ,medicine.diagnostic_test ,Infant ,PIK3R2 ,medicine.disease ,Perisylvian polymicrogyria ,3. Good health ,Malformations of Cortical Development ,mosaicism ,Child, Preschool ,Neurology (clinical) ,030217 neurology & neurosurgery - Abstract
Summary Background Bilateral perisylvian polymicrogyria (BPP), the most common form of regional polymicrogyria, causes the congenital bilateral perisylvian syndrome, featuring oromotor dysfunction, cognitive impairment, and epilepsy. The causes of BPP are heterogeneous, but only a few genetic causes have been reported. The aim of this study was to identify additional genetic causes of BPP and characterise their frequency in this population. Methods Children (aged ≤18 years) with polymicrogyria were enrolled into our research programme from July, 1980, to October, 2015, at two centres (Florence, Italy, and Seattle, WA, USA). We obtained samples (blood and saliva) throughout this period at both centres and did whole-exome sequencing on DNA from eight trios (two parents and one affected child) with BPP in 2014. After the identification of mosaic PIK3R2 mutations in two of these eight children, we performed targeted screening of PIK3R2 by two methods in a cohort of 118 children with BPP. First, we performed targeted sequencing of the entire PIK3R2 gene by single molecule molecular inversion probes (smMIPs) on 38 patients with BPP with normal to large head size. Second, we did amplicon sequencing of the recurrent PIK3R2 mutation (Gly373Arg) in 80 children with various types of polymicrogyria including BPP. One additional patient had clinical whole-exome sequencing done independently, and was included in this study because of the phenotypic similarity to our cohort. Findings We identified a mosaic mutation (Gly373Arg) in a regulatory subunit of the PI3K-AKT-mTOR pathway, PIK3R2 , in two children with BPP. Of the 38 patients with BPP and normal to large head size who underwent targeted next-generation sequencing by smMIPs, we identified constitutional and mosaic PIK3R2 mutations in 17 additional children. In parallel, one patient had the recurrent PIK3R2 mutation identified by clinical whole-exome sequencing. Seven of these 20 patients had BPP alone, and 13 had BPP in association with features of the megalencephaly-polymicrogyria-polydactyly-hydrocephalus (MPPH) syndrome. 19 patients had the same mutation (Gly373Arg), and one had a nearby missense mutation (Lys376Glu). Mutations were constitutional in 12 patients and mosaic in eight patients. In patients with mosaic mutations, we noted substantial variation in alternate (mutant) allele levels, ranging from ten (3%) of 377 reads to 39 (37%) of 106 reads, equivalent to 5–73% of cells analysed. Levels of mosaicism varied from undetectable to 37 (17%) of 216 reads in blood-derived DNA compared with 2030 (29%) of 6889 reads to 275 (43%) of 634 reads in saliva-derived DNA. Interpretation Constitutional and mosaic mutations in the PIK3R2 gene are associated with developmental brain disorders ranging from BPP with a normal head size to the MPPH syndrome. The phenotypic variability and low-level mosaicism, which challenge conventional molecular methods, have important implications for genetic testing and counselling. Funding US National Institutes of Health.
- Published
- 2015
46. Corpus callosum agenesis, severe mental retardation, epilepsy, and dyskinetic quadriparesis due to a novel mutation in the homeodomain of ARX
- Author
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William B. Dobyns, Carla Marini, Jyotsna Sudi, Simone Gana, Valerio Conti, and Renzo Guerrini
- Subjects
Male ,Acrocallosal Syndrome ,Molecular Sequence Data ,Mutation, Missense ,Quadriplegia ,medicine.disease_cause ,Young Adult ,03 medical and health sciences ,Epilepsy ,0302 clinical medicine ,Pleiotropy ,Intellectual Disability ,Genetics ,medicine ,Humans ,Missense mutation ,Amino Acid Sequence ,Agenesis of the corpus callosum ,Genetics (clinical) ,030304 developmental biology ,Homeodomain Proteins ,0303 health sciences ,Mutation ,Corpus Callosum Agenesis ,business.industry ,medicine.disease ,Pedigree ,Radiography ,Developmental disorder ,Phenotype ,Homeobox ,business ,Sequence Alignment ,030217 neurology & neurosurgery ,Transcription Factors - Abstract
We report on a patient with agenesis of the corpus callosum (ACC), severe mental retardation, infantile spasms and subsequent intractable epilepsy, spastic/dyskinetic quadriparesis, severe limb contractures, and scoliosis. This complex, newly described phenotype, is due to a novel non-conservative missense mutation in the ARX homeodomain (c.1072A>T; p.R358W), inherited from the unaffected mother. Differently from previously reported non-conservative mutations falling within the same domain, p.R358W did not cause XLAG. It is therefore possible that differences in clinical manifestations between our patient and those with XLAG, are related to the different position of the amino acid substitution in the homeodomain, or to the different chemical properties introduced by the substitution itself. To test the hypothesis that the patient's mother was asymptomatic because of non-random X chromosome inactivation (XCI), we performed DNA methylation studies of the human androgen receptor gene, demonstrating skewing of the XCI ratio (85:15). The complex phenotype described here combines different traits that had previously been linked to various ARX mutations, including conservative missense mutations in the homeodomain and expansion in the first ARX polyalanine tract and contributes to the expanding pleiotropy associated with ARX mutations.
- Published
- 2011
47. Contractions in the second polyA tract of ARX are rare, non-pathogenic polymorphisms
- Author
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Davide Mei, Valerio Conti, Carla Marini, Melania Falchi, Anna Rita Ferrari, and Renzo Guerrini
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Sequence analysis ,Molecular Sequence Data ,Biology ,Epilepsy ,Exon ,Intellectual Disability ,Gene duplication ,Genetics ,medicine ,Humans ,Child ,education ,Transcription factor ,Genetics (clinical) ,Sequence Deletion ,Homeodomain Proteins ,education.field_of_study ,Polymorphism, Genetic ,Base Sequence ,Sequence Analysis, DNA ,medicine.disease ,Phenotype ,Pedigree ,Aristaless related homeobox ,Homeobox ,Female ,Poly A ,Transcription Factors - Abstract
Aristaless related homeobox (ARX) is a transcription factor containing highly conserved octapeptide, homeobox, acidic, and aristaless domains, as well as four polyA tracts. The most frequent ARX mutation found to date in patients with X-linked infantile spasms, Partington syndrome or X-linked mental retardation, is a duplication of 24 bp in exon 2, resulting in the expansion of the second polyA tract. Although the pathogenic role of this expansion has been well characterized, the effect of contractions in the same polyA tract is still debated since different reports have associated contractions to either mental retardation or a normal phenotype. Here, we report two unrelated girls with epilepsy and mental retardation who inherited from their unaffected parents, of either sex, a deletion of 24 bp (c.441_464del), resulting in a contraction of eight alanines in the second polyA tract of ARX. Segregation studies revealed the c.441_464del also in two healthy relatives of one of the patients. This finding supports the hypothesis that this contraction represents a rare, benign polymorphism.
- Published
- 2010
48. Correction to: The landscape of epilepsy-related GATOR1 variants
- Author
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Sara Baldassari, Fabienne Picard, Nienke E. Verbeek, Marjan van Kempen, Eva H. Brilstra, Gaetan Lesca, Valerio Conti, Renzo Guerrini, Francesca Bisulli, Laura Licchetta, Tommaso Pippucci, Paolo Tinuper, Edouard Hirsch, Anne de Saint Martin, Jamel Chelly, Gabrielle Rudolf, Mathilde Chipaux, Sarah Ferrand-Sorbets, Georg Dorfmüller, Sanjay Sisodiya, Simona Balestrini, Natasha Schoeler, Laura Hernandez-Hernandez, S. Krithika, Renske Oegema, Eveline Hagebeuk, Boudewijn Gunning, Charles Deckers, Bianca Berghuis, Ilse Wegner, Erik Niks, Floor Jansen, Kees Braun, Daniëlle de Jong, Guido Rubboli, Inga Talvik, Valentin Sander, Peter Uldall, Marie-Line Jacquemont, Caroline Nava, Eric Leguern, Sophie Julia, Antonio Gambardella, Giuseppe d’Orsi, Giovanni Crichiutti, Laurence Faivre, Veronique Darmency, Barbora Benova, Pavel Krsek, Arnaud Biraben, Anne-Sophie Lebre, Mélanie Jennesson, Shifteh Sattar, Cécile Marchal, Douglas R. NordliJr, Kristin Lindstrom, Pasquale Striano, Lysa Boissé Lomax, Courtney Kiss, Fabrice Bartolomei, Anne Fabienne Lepine, An-Sofie Schoonjans, Katrien Stouffs, Anna Jansen, Eleni Panagiotakaki, Brigitte Ricard-Mousnier, Julien Thevenon, Julitta de Bellescize, Hélène Catenoix, Thomas Dorn, Martin Zenker, Karen Müller-Schlüter, Christian Brandt, Ilona Krey, Tilman Polster, Markus Wolff, Meral Balci, Kevin Rostasy, Guillaume Achaz, Pia Zacher, Thomas Becher, Thomas Cloppenborg, Christopher J. Yuskaitis, Sarah Weckhuysen, Annapurna Poduri, Johannes R. Lemke, Rikke S. Møller, Stéphanie Baulac, Reproduction and Genetics, Clinical sciences, Medical Genetics, Mental Health and Wellbeing research group, Public Health Sciences, Neurogenetics, and Pediatrics
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ComputingMethodologies_DOCUMENTANDTEXTPROCESSING ,Correction ,Genetics(clinical) ,Genetics (clinical) - Abstract
The original version of this Article contained an error in the author list where the corresponding author Stéphanie Baulac was repeated twice. This has now been corrected in the HTML, the PDF was correct at the time of publication.
- Published
- 2018
49. Computer-based automatic identification of neurons in gigavoxel-sized 3D human brain images
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Paolo Frasconi, Renzo Guerrini, Irene Costantini, Francesco S. Pavone, Giulio Iannello, Paolo Soda, Ermanno Cordelli, Ludovica Acciai, Valerio Conti, and Leonardo Sacconi
- Subjects
Neurons ,Nervous system ,Hemimegalencephaly ,biology ,business.industry ,Brain ,Neuroimaging ,Human brain ,Macaque ,Imaging, Three-Dimensional ,medicine.anatomical_structure ,Cytoarchitecture ,biology.animal ,medicine ,Fluorescence microscope ,Humans ,Soma ,business ,Neuroscience - Abstract
Achieving a comprehensive knowledge of the human brain cytoarchitecture is a fundamental step to understand how the nervous system works, i.e., one of the greatest challenge of 21(st) century science. The recent development of biological tissue labeling and automated microscopic imaging systems has permitted to acquire images at the micro-resolution, which produce a huge quantity of data that cannot be manually analyzed. In case of mammals brain, automatic methods to extract objective information at the microscale have been applied until now to mice, macaque and cat 3D volume images. Here we report a method to automatically localize neurons in a sample of human brain removed during a surgical procedure for the treatments of drug resistant epilepsy in a child with hemimegalencephaly, whose neurons and neurites were fluorescence labelled and finally imaged using the two-photon fluorescence microscope. The method provides the map of both parvalbuminergic neurons and all other cells nuclei with a satisfactory f-score measured using more than two thousand human labelled soma.
- Published
- 2015
50. A versatile clearing agent for multi-modal brain imaging
- Author
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Francesco Vanzi, Leonardo Sacconi, Renzo Guerrini, Ludovico Silvestri, Jean Pierre Ghobril, Anna Letizia Allegra Mascaro, Francesco S. Pavone, Marie Caroline Müllenbroich, Giulio Iannello, Irene Costantini, Valerio Conti, Henry Markram, Antonino Paolo Di Giovanna, and Leonardo Onofri
- Subjects
Computer science ,Contrast Media ,FOS: Physical sciences ,brain imaging ,Neuroimaging ,Brain tissue ,Article ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Animals ,Humans ,Clearing Agent ,Tomography ,030304 developmental biology ,light sheet microscopy ,Mouse Hippocampus ,0303 health sciences ,Multidisciplinary ,Modality (human–computer interaction) ,serial two-photon microscopy ,Brain ,Physics - Medical Physics ,Immunohistochemistry ,clearing ,Modal ,Quantitative Biology - Neurons and Cognition ,FOS: Biological sciences ,Light sheet fluorescence microscopy ,Neurons and Cognition (q-bio.NC) ,Medical Physics (physics.med-ph) ,030217 neurology & neurosurgery ,Biomedical engineering - Abstract
Extensive mapping of neuronal connections in the central nervous system requires high-throughput um-scale imaging of large volumes. In recent years, different approaches have been developed to overcome the limitations due to tissue light scattering. These methods are generally developed to improve the performance of a specific imaging modality, thus limiting comprehensive neuroanatomical exploration by multimodal optical techniques. Here, we introduce a versatile brain clearing agent (2,2'-thiodiethanol; TDE) suitable for various applications and imaging techniques. TDE is cost-efficient, water-soluble and low-viscous and, more importantly, it preserves fluorescence, is compatible with immunostaining and does not cause deformations at sub-cellular level. We demonstrate the effectiveness of this method in different applications: in fixed samples by imaging a whole mouse hippocampus with serial two-photon tomography; in combination with CLARITY by reconstructing an entire mouse brain with light sheet microscopy and in translational research by imaging immunostained human dysplastic brain tissue., Comment: in Scientific Reports 2015
- Published
- 2015
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