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International audience; Congenital limb malformations (CLM) comprise many conditions affecting limbs and more than 150 associated genes have been reported. Due to this large heterogeneity, a high proportion of patients remains without a molecular diagnosis. In the last two decades, advances in high throughput sequencing have allowed new methodological strategies in clinical practice. Herein, we report the screening of 52 genes/regulatory sequences by multiplex high-throughput targeted sequencing, in a series of 352 patients affected with various CLM, over a 3-year period of time. Patients underwent a clinical triage by expert geneticists in CLM. A definitive diagnosis was achieved in 35.2% of patients, the yield varying considerably, depending on the phenotype. We identified 112 single nucleotide variants and 26 copy-number variations, of which 52 are novel pathogenic or likely pathogenic variants. In 6% of patients, variants of uncertain significance have been found in good candidate genes. We showed that multiplex targeted high-throughput sequencing works as an efficient and cost-effective tool in clinical practice for molecular diagnosis of congenital limb malformations. Careful clinical evaluation of patients may maximize the yield of CLM panel testing.

International audience; Chemical burns are often deep with difficult initial clinical evaluation, especially those due to alkalic agents, which have a strong penetrating power. They therefore require specialized care in a Burn Unit. Self-inflicted burns are infrequent but their management represents a real challenge. We report the case of a 47-year-old referred to our Burn Center for the evaluation of a self-inflicted corrosion with an alkalic agent (soda), injected at the crease of the left elbow. The patient, right handed, was a nurse and had notably a psychiatric history of depressive syndrome. We observed a deep, well-defined necrosis area, associated with intense peri-lesional inflammation and extensive cellulitis. Faced with this unusual clinical appearance for a chemical burn, the patient's questioning was repeated and the patient finally admitted to having injected himself with a basic caustic product intravenously. Surgical treatment was carried out in two stages: debridement with exposure of vascular and neural structures then coverage with a free anterolateral thigh flap. The postoperative consequences were uneventful with a satisfactory functional result. Factitious disorders are underestimated and often misleading. Among factitious disorders, self-inflicted wounds remain a real challenge requiring multidisciplinary management. Many etiologies exist, among which injection of drugs or substances, in any anatomical localization, leading to variable loss of substance. The use of a free flap for acute extravasation is rare but sometimes essential. The anterolateral thigh flap allows good resurfacing on areas with important functional requirements.; Les brûlures chimiques sont souvent des brûlures profondes, dont l’évaluation clinique initiale est difficile, en particulier pour les brûlures causées par des agents chimiques basiques, ayant un fort pouvoir pénétrant. Elles nécessitent une prise en charge spécialisée dans un centre de traitement des brûlés. Les brûlures auto-infligées sont peu fréquentes et leur prise en charge représente un véritable challenge. Nous rapportons le cas d’un patient de 47 ans adressé initialement dans notre centre pour une brûlure chimique basique du pli du coude gauche suite à un contact accidentel avec de la soude. Le patient droitier et infirmier présentait, notamment comme antécédent, un syndrome dépressif. Nous avons observé une zone nécrotique bien délimitée au niveau du pli du coude associée à une inflammation péri-lésionnelle intense. Devant cet aspect inhabituel pour une brûlure chimique par contact, nous avons répété l’interrogatoire du patient. Ce dernier a fini par reconnaître s’être auto-infligé une injection intraveineuse de soude au niveau du pli du coude. Un traitement chirurgical en deux étapes a été réalisé : parage avec exposition de structures vasculonerveuses, puis couverture par un lambeau antérolatéral de cuisse libre. Les suites postopératoires ont été simples avec un résultat fonctionnel satisfaisant. Les « troubles factices » parmi lesquels les automutilations restent un véritable défi nécessitant une prise en charge multidisciplinaire. Les pertes de substance sont de taille et de profondeur variables. L’utilisation des lambeaux libres dans les cas d’extravasation est rare, mais parfois indispensable. Le lambeau antérolatéral de cuisse permet un bon resurfaçage des zones fonctionnelles.

Summary Background & aims Mitochondrial OXPHOS disorders (MODs) affect one or several complexes of respiratory chain oxidative phosphorylation. An increased fat/low-carbohydrate ratio of the diet was recommended for treating MODs without, however, evaluating its potential benefits through changes in the respective contributions of cell pathways (glycolysis, fatty acid oxidation) initiating energy production. Therefore, the objective of the present work was to compare Resting Energy Expenditure (REE) under basal diet (BD) and challenging diet (CD) in which fat on sugar content ratio was doubled. Diet-induced thermogenesis (DIT) and body compositions were also compared. Energetic vs regulatory aspects of increasing fat contribution to total nutritional energy input were essentially addressed through measures primarily aiming at modifying total fat amounts and not the types of fats in designed diets. Methods In this randomized cross-over study, BD contained 10% proteins/30% lipids/60% carbohydrates (fat on sugar ratio = 0.5) and was the imposed diet at baseline. CD contained 10% proteins/45% lipids/45% carbohydrates (fat on sugar ratio = 1). Main and second evaluation criteria measured by indirect calorimetry (QUARK RMR ® , Cosmed, Pavona; Italy) were REE and DIT, respectively. Thirty four MOD patients were included; 22 (mean age 13.2 ± 4.7 years, 50% female; BMI 16.9 ± 4.2 kg/m 2 ) were evaluated for REE, and 12 (mean age 13.8 ± 4.8 years, 60% female; BMI 17.4 ± 4.6 kg/m 2 ) also for DIT. OXPHOS complex deficiency repartition in 22 analysed patients was 55% for complex I, 9% for complex III, 27% for complex IV and 9% for other proteins. Results Neither carry-over nor period effects were detected (p = 0.878; ANOVA for repeated measures). REE was similar between BD vs CD (1148.8 ± 301.7 vs 1156.1 ± 278.8 kcal/day; p = 0.942) as well as DIT (peak DIT 260 vs 265 kcal/day; p = 0.842) and body composition (21.9 ± 13.0 vs 21.6 ± 13.3% of fat mass; p = 0.810). Conclusion Doubling diet fat on sugar ratio does not appear to improve, per se , energetic status and body composition of patients with MODs.

International audience; Silver-Russell syndrome (SRS) is a rare imprinting disorder characterized by prenatal and postnatal growth retardation. The two principal causes of SRS are loss of methylation on chromosome 11p15 (11p15 LOM) and maternal uniparental disomy of chromosome 7 (UPD(7)mat). Knowledge of the neuropsychological profile of SRS remains sparse and incomplete even if several difficulties related to attention and learning have been reported both in the literature and by patients with SRS. These difficulties could be the result of troubles in different cognitive domains, but also of executive dysfunction. Nevertheless, executive functioning has never been investigated, even though executive functions play an essential role in psychological development, and are extensively involved in daily life. The present study explored the executive functioning of individuals with SRS due to UPD(7)mat or 11p15 LOM. A battery of executive tasks assessing cognitive flexibility, inhibitory control, and working memory, together with a task assessing sustained attention, was administered to 19 individuals with SRS (13–39 years) and 19 healthy controls. The Behavior Rating Inventory of Executive Function was also completed by the participants’ families. The results showed that participants with SRS had similar performance (z-scores) to our controls, in a context of normal intellectual efficiency. Group comparisons with Bayesian statistics showed a single difference between the 11p15 LOM and control groups: the completion time for part A of the Trail Making Test appeared to be longer in the 11p15 LOM group than in the control group. However, at the clinical level, several participants with SRS had clinically significant scores on various measures of EFs. Thus, the cognitive phenotype of SRS did not appear to be characterized by executive dysfunction, but individuals with SRS could be at high risk of developing executive dysfunction or attention-deficit/hyperactivity disorder. These results provide new insights into the neuropsychological profile of individuals with SRS.

International audience; Background:Infectious purpura fulminans is a disabling disease often leading to amputations. Free flaps preserve limb length, covering exposed areas. We examined the efficacy of free flaps for lower limb salvage in infectious purpura fulminans survivors by evaluating surgical management, walking ability and quality of life.Methods:This single-center, observational, descriptive, retrospective study was conducted in from 2016 to 2019. Adult purpura fulminans survivors who received a free flap for lower limb salvage were included. Patient characteristics and data on surgical management and rehabilitation were collected. Quality of life (SF-36 questionnaire), limb function and walking ability were later evaluated post-surgically.Results:The 6 patients included, mean age 38 years, had all required amputations. Nine free flaps were performed to cover important structures in 7 cases and for stump resurfacing in 2. All flaps were successful. Patients resumed walking at a mean of 204 ± 108 days after the onset of purpura fulminans. Post-surgical evaluation was performed at a mean of 30 ± 9.3 months. Five patients required secondary revision. All were independent for the activities of daily living. Mean physical component score was 37.6 ± 9.4 and mental component score was 44.6 ± 13.2 (minimum 0, maximum 100).Conclusions:Use of the free flap in patients with infectious purpura fulminans, after multidisciplinary reflection, is an appropriate procedure that preserves limb length. In spite of secondary complications, preservation of limb length enables patients to resume walking, with relatively good independence and quality of life.; Introduction :Le purpura fulminans infectieux est une pathologie invalidante entraînant souvent des amputations. L’objectif est d’évaluer le sauvetage de membre inférieur par lambeaux libres dans le purpura fulminans infectieux : la prise en charge chirurgicale, la marche, et la qualité de vie des patients.Méthodes :Il s’agit d’une étude observationnelle, descriptive, rétrospective de 2016 à 2019. Les patients inclus étaient des adultes survivants au purpura fulminans, bénéficiant d’un lambeau libre pour sauvetage d’un membre inférieur. Les données recueillies étaient les caractéristiques des patients, la prise en charge chirurgicale et la rééducation. L’évaluation à distance portait sur les patients, la marche, leur qualité de vie.Résultats :Six patients ont été inclus, avec 38 ans de moyenne d’âge. Neuf lambeaux libres ont été réalisés, et se sont déroulés avec succès. La reprise de la marche s’est effectuée en moyenne 204 ± 108 jours après le purpura fulminans. L’évaluation à distance était en moyenne réalisée à 30 ± 9,3 mois. Cinq patients sur 6 ont nécessité des corrections secondaires. Tous les patients sont autonomes dans la vie quotidienne. Le score résumé physique était en moyenne de 37,6 ± 9,4, le score résumé psychique de 44,6 ± 13,2 (minimum 0, maximum 100).Conclusion :Le lambeau libre chez les patients atteints de purpura fulminans infectieux est une procédure adaptée, permettant de préserver la longueur de membre, avec un bénéfice pour le patient malgré les complications secondaires, permettant une reprise de la marche, une autonomie, et une qualité de vie relativement bonne.

International audience; Developmental Cerebellar Anomalies (DCA) are rare diseases (e.g., Joubert syndrome) that affect various motor and non-motor functions during childhood. The present study examined whether music perception and production are affected in children with DCA. Sixteen children with DCA and 37 healthy matched control children were tested with the Montreal Battery for Evaluation of Musical Abilities (MBEMA) to assess musical perception. Musical production was assessed using two singing tasks: a pitch-matching task and a melodic reproduction task. Mixed model analyses showed that children with DCA were impaired on the MBEMA rhythm perception subtest, whereas there was no difference between the two groups on the melodic perception subtest. Children with DCA were also impaired in the melodic reproduction task. In both groups, singing performance was positively correlated with rhythmic and melodic perception scores, and a strong correlation was found between singing ability and oro-bucco-facial praxis in children with DCA. Overall, children with DCA showed impairments in both music perception and production, although heterogeneity in cerebellar patient's profiles was highlighted by individual analyses. These results confirm the role of the cerebellum in rhythm processing as well as in the vocal sensorimotor loop in a developmental perspective. Rhythmic deficits in cerebellar patients are discussed in light of recent work on predictive timing networks including the cerebellum. Our results open innovative remediation perspectives aiming at improving perceptual and/or production musical abilities while considering the heterogeneity of patients' clinical profiles to design music-based therapies.

International audience; Craniostenosis is a morphological anomaly affecting about 0.5 of 1000 births and one third of the cases are of genetic origin. Among the syndromes responsible for craniostenosis, there is the Saethre-Chotzen syndrome due to a mutation of the TWIST 1 gene located on chromosome 7. This polymalformative syndrome classically includes a particular morphology of the auricles. The penetrance is variable and results in a phenotypic variability at the origin of “Saethre-Chotzen like” clinical pictures for which the TWIST 1 gene mutation is sometimes not found. Recently, the TCF 12 gene has been implicated in some of these cases. Among the multiple facial malformations, we have carefully examined the particular morphology of the auricle of these patients. The authors found several abnormalities in patients with a TCF 12 gene mutation, namely a thickened and hammered upper pole of the helix, a narrow concha without crux cymbae and a thickened lobe. These morphological features may guide the diagnosis and allow an earlier search for a TCF 12 gene mutation.

International audience; No abstract available

International audience; Several techniques for the stabilization and the reanimation of the paralyzed lower lip have been proposed, sometimes combined but the authors are rather vague. Thanks to our cases and a literature review, we suggest an algorithm. Sixty-six patients haven been reviewed. The static and dynamic clinical evaluation have been done thanks to the study of the medical files, photographs and videos samples. The different techniques we have used were: (a) botulinum toxin in the contralateral depressor labii inferioris (DLI) (n = 66), contralateral DLI myectomy (n = 1); (b) a strip of fascia lata (n = 16) either combined with a lengthening temporalis myoplasty (MAT) (n = 10) or only realizing a passive suspension thanks to one vector (n = 4), or a passive suspension with two strips (n = 2); (c) digastric muscle transfer (n = 4). When the elevator muscles are sounds, botulin toxin (± myectomy) is proposed if the lower lip is not inverted. Otherwise, a suspension thanks to a digastric muscle transfer or a passive suspension with a horizontal and a vertical strips are recommended. If the elevator muscles are no more operational, a MAT with a passive suspension with one vector is considerated and if the lower lip is inverted, a digastric muscle transfer with a fascia lata strip or a MAT with two strips is proposed. This algorithm submission allows to refine the treatment of the stabilization and the reanimation of the lower lip.; De multiples approches thérapeutiques de la stabilisation et réanimation de la lèvre inférieure paralysée ont été proposées, parfois combinées mais les auteurs restent peu précis. Nous proposons à travers nos cas et la littérature un algorithme. Soixante-six patients ont été revus rétrospectivement. L’évaluation clinique statique et dynamique a été effectuée par étude de dossiers, photographies et enregistrements vidéographiques. Les différents moyens thérapeutiques utilisés ont été : (a) la toxine botulique dans le depressor labii inferioris (DLI) controlatéral (n = 66), la myectomie du DLI controlatéral (n = 1) ; (b) une bandelette de fascia lata (n = 16) soit connectée à une myoplastie d’allongement du temporal (MAT) (n = 10), soit réalisant une suspension passive par un seul vecteur (n = 4), soit réalisant une suspension passive par 2 vecteurs (n = 2) ; (c) un transfert du ventre postérieur du muscle digastrique (n = 4). Quand les muscles élévateurs sont sains, la toxine botulique (± myectomie) est proposée si la lèvre inférieure n’est pas inversée. Dans le cas contraire, une suspension par transfert du digastrique ou une suspension passive à deux vecteurs est à prévoir. Si les muscles releveurs ne sont plus fonctionnels, il faut envisager une MAT avec une suspension passive à un vecteur et si la lèvre est inversée, un transfert du muscle digastrique avec une bandelette de fascia lata ou une MAT associé à deux bandelettes de fascia lata. Cette proposition d’algorithme permet de raffiner le traitement de la stabilisation et de la réanimation de la lèvre inférieure.

International audience; Contexte: Le déficit en alpha 1-antitrypsine (DAAT) est une maladie génétique autosomique récessive associée à l’état homozygote du variant Z du gène SERPINA1. Les manifestations cliniques sont un déficit sévère en alpha 1-antitrypsine, un emphysème pulmonaire et une fibrose hépatique.Méthodes: Des recommandations de prise en charge du DAAT ont été élaborées à l’initiative du centre coordonnateur de référence des maladies pulmonaires rares, sous l’égide de la Société de pneumologie de langue française, par des groupes de coordination, rédaction, et lecture, impliquant des pneumologues de divers modes d’exercice, biologistes, hépatologue, pharmacien hospitalier, conseiller en génétique, radiologues, médecins généralistes, cadre de santé, et associations de patients. La méthode d’élaboration des « Recommandations pour la pratique clinique » de la Haute autorité de santé a été suivie, incluant un vote en ligne selonune échelle Likert.Résultats: Après une analyse bibliographique, 20 recommandations ont été proposées par les groupes de travail, portant sur tous les aspects de la maladie : diagnostic biologique, bilan initial, conseils d’hygiène de vie, information, indications et modalités du traitement substitutif, dépistage.Conclusion: Ces recommandations fondées sur les preuves sont destinées à guider le diagnostic et la prise en charge pratique du DAAT.

Oxidative stress and inflammation are the key players in neuroinflammation, in which microglia dysfunction plays a central role. Previous studies suggest that argan oil attenuates oxidative stress, inflammation, and peroxisome dysfunction in mouse brains. In this study, we explored the effects of two major argan oil (AO) phytosterols, Schottenol (Schot) and Spinasterol (Spina), on oxidative stress, inflammation, and peroxisomal dysfunction in two murine microglial BV-2 cell lines, wild-ype (Wt) and Acyl-CoA oxidase 1 (Acox1)-deficient cells challenged with LPS treatment. Herein, we used an MTT test to reveal no cytotoxicity for both phytosterols with concentrations up to 5 µM. In the LPS-activated microglial cells, cotreatment with each of these phytosterols caused a significant decrease in intracellular ROS production and the NO level released in the culture medium. Additionally, Schot and Spina were able to attenuate the LPS-dependent strong induction of Il-1β and Tnf-α mRNA levels, as well as the iNos gene and protein expression in both Wt and Acox1−/− microglial cells. On the other hand, LPS treatment impacted both the peroxisomal antioxidant capacity and the fatty acid oxidation pathway. However, both Schot and Spina treatments enhanced ACOX1 activity in the Wt BV-2 cells and normalized the catalase activity in both Wt and Acox1−/− microglial cells. These data suggest that Schot and Spina can protect cells from oxidative stress and inflammation and their harmful consequences for peroxisomal functions and the homeostasis of microglial cells. Collectively, our work provides a compelling argument for the protective mechanisms of two major argan oil phytosterols against LPS-induced brain neuroinflammation.

Pathogenic heterozygous MED13L variants cause a rare neurodevelopmental disorder characterized by a moderate-to-severe intellectual disability. Whole exome or genome sequencing in patients with non-specific neurodevelopmental disorders leads to the identification of an increasing number of MED13L candidate missense variations.Through international collaboration between centers of expertise for developmental anomalies, we report 36 patients with MED13L molecular anomaly to expand the phenotype. Five MED13L pathogenic missense variants causing either a severe phenotype (p.Pro866Leu, p.Pro869Ser, and p.Cys1131Tyr) or a typical phenotype (p.Gly1899Arg and p.Thr2162Met) were selected and functionally studied. Along with the functional analysis, a comparison of in silico parameters associated either with previously published pathogenic missense or neutral missenses was performed.Missense variants clustered in two mutation hot-spots, exons 15–17 and 25–31. We found that patients carrying missense variations showed a more severe phenotype with frequent epilepsy. Significant differences were identified between pathogenic and neutral missenses for global conservation, physico-chemical difference between amino acids by Grantham scores, conservation of sequence in MED13 protein, and proximity to phosphorylation sites. Through functional evaluations, the p.Pro866Leu and p.Pro869Ser variants were likely to induce a hyper-phosphorylated status of the MED13L823-930 domain. Residues Cys1131 and Thr2162 are predicted to be critical for MED13L proper folding, and substitutions are likely to induce modification of the 3D structure. Increased proteasome-mediated degradation, was observed only with the p.Gly1899Arg variant as well as cytoplasmic relocalization and reduced integration into CKM and core Mediator complex.This study ascertains missense variations in MED13L as a cause for MED13L-related intellectual disability and improves the clinical and molecular delineations of the condition. MED13L variants associated with typical phenotypes (p.Gly1899Arg and p.Thr2162Met) probably induce a loss of function, while variants associated with severe phenotypes probably cause a dominant-negative effect., Les variations hétérozygotes pathogènes de MED13L provoquent un trouble du neurodéveloppement rare caractérisé par une déficience intellectuelle modérée à sévère. Le séquençage non ciblé chez des patients atteints de déficiences intellectuelles non-syndromiques a conduit à l'identification d'un nombre croissant de variations faux-sens candidates dans MED13L.Par une collaboration internationale entre des centres d'expertise pour les anomalies du développement, nous avons rapporté 36 patients présentant une variation pathogène de MED13L afin de préciser le phénotype. Cinq variantes faux-sens pathogènes provoquant soit un phénotype sévère (p.Pro866Leu, p.Pro869Ser, et p.Cys1131Tyr) soit un phénotype typique (p.Gly1899Arg et p.Thr2162Met) ont été sélectionnées et étudiées fonctionnellement. Parallèlement à l'analyse fonctionnelle, une comparaison des paramètres in silico associés aux faux-sens pathogènes précédemment publiés ou rapportées dans les bases de données témoins a été réalisée.Les variants faux-sens sont regroupés majoritairement dans les exons 15-17 et 25-31. Nous avons constaté que les patients porteurs de variations faux-sens présentaient un phénotype plus sévère avec une épilepsie fréquente. Des différences significatives ont été identifiées entre les faux-sens pathogènes et neutres pour les scores de conservation, les différences physico-chimiques entre acides aminés suivant le score de Grantham, la conservation de la séquence dans la protéine MED13 et la proximité de sites de phosphorylation. Fonctionnement, les variations p.Pro866Leu et p.Pro869Ser étaient susceptibles d'induire une hyper-phosphorylation du domaine MED13L823-930. Les résidus Cys1131 et Thr2162 pourraient impacter la conformation de MED13L avec une modification de la structure 3D. Une dégradation accrue médiée par le protéasome ainsi qu’une relocalisation cytoplasmique et une modification de l’intégration dans le module kinase CDK8 ont été observées uniquement pour le faux-sens p.Gly1899Arg.Cette étude confirme que les variations faux-sens de MED13L sont une cause du Syndrome MED13L avec des particularités cliniques et moléculaires. Les faux-sens de MED13L associées aux phénotypes typiques (p.Gly1899Arg et p.Thr2162Met) induisent probablement une perte de fonction, tandis que les variations associées aux phénotypes sévères provoqueraient probablement un effet dominant-négatif.

Background In France, the Ministry of Health has implemented a comprehensive program for rare diseases (RD) that includes an epidemiological program as well as the establishment of expert centers for the clinical care of patients with RD. Since 2007, most of these centers have entered the data for patients with developmental disorders into the CEMARA population-based registry, a national online data repository for all rare diseases. Through the CEMARA web portal, descriptive demographic data, clinical data, and the chronology of medical follow-up can be obtained for each center. We address the interest and ongoing challenges of this national data collection system 10 years after its implementation. Methods Since 2007, clinicians and researchers have reported the “minimum dataset (MDS)” for each patient presenting to their expert center. We retrospectively analyzed administrative data, demographic data, care organization and diagnoses. Results Over 10 years, 228,243 RD patients (including healthy carriers and family members for whom experts denied any suspicion of RD) have visited an expert center. Among them, 167,361 were patients affected by a RD (median age 11 years, 54% children, 46% adults, with a balanced sex ratio), and 60,882 were unaffected relatives (median age 37 years). The majority of patients (87%) were seen no more than once a year, and 52% of visits were for a diagnostic procedure. Among the 2,869 recorded rare disorders, 1,907 (66.5%) were recorded in less than 10 patients, 802 (28%) in 10 to 100 patients, 149 (5.2%) in 100 to 1,000 patients, and 11 (0.4%) in > 1,000 patients. Overall, 45.6% of individuals had no diagnosis and 6.7% had an uncertain diagnosis. Children were mainly referred by their pediatrician (46%; n = 55,755 among the 121,136 total children referrals) and adults by a medical specialist (34%; n = 14,053 among the 41,564 total adult referrals). Given the geographical coverage of the centers, the median distance from the patient’s home was 25.1 km (IQR = 6.3 km-64.2 km). Conclusions CEMARA provides unprecedented support for epidemiological, clinical and therapeutic studies in the field of RD. Researchers can benefit from the national scope of CEMARA data, but also focus on specific diseases or patient subgroups. While this endeavor has been a major collective effort among French RD experts to gather large-scale data into a single database, it provides tremendous potential to improve patient care.

BackgroundVariants in HECW2 have recently been reported to cause a neurodevelopmental disorder with hypotonia, seizures and impaired language; however, only six variants have been reported and the clinical characteristics have only broadly been defined.MethodsMolecular and clinical data were collected from clinical and research cohorts. Massive parallel sequencing was performed and identified individuals with a HECW2-related neurodevelopmental disorder.ResultsWe identified 13 novel missense variants in HECW2 in 22 unpublished cases, of which 18 were confirmed to have a de novo variant. In addition, we reviewed the genotypes and phenotypes of previously reported and new cases with HECW2 variants (n=35 cases). All variants identified are missense, and the majority of likely pathogenic and pathogenic variants are located in or near the C-terminal HECT domain (88.2%). We identified several clustered variants and four recurrent variants (p.(Arg1191Gln);p.(Asn1199Lys);p.(Phe1327Ser);p.(Arg1330Trp)). Two variants, (p.(Arg1191Gln);p.(Arg1330Trp)), accounted for 22.9% and 20% of cases, respectively. Clinical characterisation suggests complete penetrance for hypotonia with or without spasticity (100%), developmental delay/intellectual disability (100%) and developmental language disorder (100%). Other common features are behavioural problems (88.9%), vision problems (83.9%), motor coordination/movement (75%) and gastrointestinal issues (70%). Seizures were present in 61.3% of individuals. Genotype-phenotype analysis shows that HECT domain variants are more frequently associated with cortical visual impairment and gastrointestinal issues. Seizures were only observed in individuals with variants in or near the HECT domain.ConclusionWe provide a comprehensive review and expansion of the genotypic and phenotypic spectrum of HECW2 disorders, aiding future molecular and clinical diagnosis and management.

The X‐linked PTCHD1 gene, encoding a synaptic membrane protein, has been involved in neurodevelopmental disorders with the description of deleterious genomic microdeletions or truncating coding mutations. Missense variants were also identified, however, without any functional evidence supporting their pathogenicity level. We investigated 13 missense variants of PTCHD1, including eight previously described (c.152G>A,p.(Ser51Asn); c.217C>T,p.(Leu73Phe); c.517A>G,p.(Ile173Val); c.542A>C,p.(Lys181Thr); c.583G>A,p.(Val195Ile); c.1076A>G,p.(His359Arg); c.1409C>A,p.(Ala470Asp); c.1436A>G,p.(Glu479Gly)), and five novel ones (c.95C>T,p.(Pro32Leu); c.95C>G,p.(Pro32Arg); c.638A>G,p.(Tyr213Cys); c.898G>C,p.(Gly300Arg); c.928G>C,p.(Ala310Pro)) identified in male patients with intellectual disability (ID) and/or autism spectrum disorder (ASD). Interestingly, several of these variants involve amino acids localized in structural domains such as transmembrane segments. To evaluate their potentially deleterious impact on PTCHD1 protein function, we performed in vitro overexpression experiments of the wild‐type and mutated forms of PTCHD1‐GFP in HEK 293T and in Neuro‐2a cell lines as well as in mouse hippocampal primary neuronal cultures. We found that six variants impaired the expression level of the PTCHD1 protein, and were retained in the endoplasmic reticulum suggesting abnormal protein folding. Our functional analyses thus provided evidence of the pathogenic impact of missense variants in PTCHD1, which reinforces the involvement of the PTCHD1 gene in ID and in ASD., Thirteen missense variants in the PTCHD1 gene associated with X‐linked neurodevelopmental disorder were investigated to address their potentially deleterious impact. Their overexpression in various cell lines (HEK293T, Neuro‐2a) and in primary neuronal cultures revealed abnormal protein stability and impaired subcellular localization, thus providing further evidence for the significant impact of PTCHD1 genetic alterations in intellectual disability and in autism spectrum disorder.

International audience; Owing to their ease of use, glucose meters are frequently used in research and medicine. However, little is known of whether other non-glucose molecules, besides vitamin C, interfere with glucometry. Therefore, we sought to determine whether other antioxidants might behave like vitamin C in causing falsely elevated blood glucose levels, potentially exposing patients to glycemic mismanagement by being administered harmful doses of glucose-lowering drugs. To determine whether various antioxidants can be detected by seven commercial glucose meters, human blood samples were spiked with various antioxidants ex vivo and their effect on the glucose results were assessed by Parkes error grid analysis. Several of the glucose meters demonstrated a positive bias in the glucose measurement of blood samples spiked with vitamin C, N-acetylcysteine, and glutathione. With the most interference-sensitive glucose meter, non-blood solutions of 1 mmol/L N-acetylcysteine, glutathione, cysteine, vitamin C, dihydrolipoate, and dithiothreitol mimicked the results seen on that glucose meter for 0.7, 1.0, 1.2, 2.6, 3.7 and 5.5 mmol/L glucose solutions, respectively. Glucose meter users should be alerted that some of these devices might produce spurious glucose results not only in patients on vitamin C therapy but also in those being administered other antioxidants. As discussed herein, the clinical relevance of the data is immediate in view of the current use of antioxidant therapies for disorders such as the metabolic syndrome, diabetes, cardiovascular diseases, and coronavirus disease 2019.

Nail-Patella syndrome (NPS) is a genetic disorder generating physical malformations and, in approximately one in three cases, ocular and renal damage. The present research aimed to deeply understand patients' subjective experience with NPS, particularly the aspects of the syndrome that affect patients' adaptation and to propose interventions that can improve genetic and psychological counseling and help patients cope with their condition. Semi-structured interviews of nine people diagnosed with NPS were analyzed using interpretative phenomenological analysis. Results highlighted attempts to look like a person without disabilities by hiding malformations and not telling the truth about symptoms' genetic origin because of patients' poor self-esteem, negative self-cognition, and social isolation experienced from childhood to adulthood. Difficulties of adaptation to physical limits and pain were also identified. The majority of participants who were not diagnosed at birth tended to consider physical symptoms as “birth malformations” without imagining other potential implications until receiving a diagnosis. Despite the diagnosis, the majority continued to minimize the potential complications by considering NPS as a “physical difference” and not adhering to medical surveillance., SCOPUS: ar.j, info:eu-repo/semantics/published

Background The Registry of Adult and Paediatric Patients Treated with Cystadane® – Homocystinuria (RoCH) is a non-interventional, observational, multi-centre, post-authorization safety study that aimed to identify safety of betaine anhydrous (Cystadane®) in the treatment of patients with inborn errors of homocysteine metabolism (homocystinuria) in order to minimise the treatment associated risks and establish better knowledge on its clinical use. The registry included patients of all ages with homocystinuria who were treated with betaine anhydrous in conjunction with other therapies. Clinical data were collected retrospectively from 2007 to 2013, then prospectively up to February 2014. All adverse events (AEs) reported during the study were recorded. The clinical and biological status of patients was monitored at least once a year. Results A total of 125 patients with homocystinuria (adults [> 18 years]: 50; paediatric [≤18 years]: 75) were enrolled at 29 centres in France and Spain. Patients were treated with betaine anhydrous for a mean duration of 7.4 ± 4.3 years. The median total daily dose of betaine anhydrous at the first and last study visits was 6 g/day for cystathionine β-synthase (CBS)-deficient vitamin B6 responders and 9 g/day for methylenetetrahydrofolate reductase-deficient patients, while the median daily dose increased in CBS-deficient B6 non-responders (from 6 to 9 g/day) and cobalamin metabolism-defective patients (from 3 to 6 g/day) between the first and last visits. Treatment caused a mean overall reduction of 29% in plasma homocysteine levels in the study population. A total of 277 AEs were reported during the study, of which two non-serious AEs (bad taste and headache) and one serious AE (interstitial lung disease) were considered to be drug related. Overall, betaine anhydrous was well tolerated with no major safety concerns. Conclusions Data from the RoCH registry provided real-world evidence on the clinical safety and efficacy of betaine anhydrous in the management of homocystinuria in paediatric and adult patients. Electronic supplementary material The online version of this article (10.1186/s13023-019-1036-2) contains supplementary material, which is available to authorized users.

International audience; Children with a written language disorder are sometimes dependent upon help from others for their schoolwork. A computer can be a way to circumvent this difficulty. Various software programs and plug-in peripheral devices are available, some of which specifically target the needs of these young people. There is no consensus, however, with regard to how best to counsel parents and children with regard to these tools. Furthermore, written language disorders and existing technical supports are not always clearly understood. In many cases, healthcare and teaching professionals have only limited knowledge of the potentially specific advantages for patients with written language disorders. A child's full integration into daily activities and school life can be hampered by counseling that was inadequately tailored or by a lack of support in using this equipment. Joint consultations involving both an occupational and a speech therapist have been set up in our department to improve counseling with regard to technical supports. Using our daily practice as a basis, we have developed a decision tree that we see as a necessary tool for helping professionals make the most appropriate practical choices.

RLIM, also known as RNF12, is an X-linked E3 ubiquitin ligase acting as a negative regulator of LIM-domain containing transcription factors and participates in X-chromosome inactivation (XCI) in mice. We report the genetic and clinical findings of 84 individuals from nine unrelated families, eight of whom who have pathogenic variants in RLIM (RING finger LIM domain-interacting protein). A total of 40 affected males have X-linked intellectual disability (XLID) and variable behavioral anomalies with or without congenital malformations. In contrast, 44 heterozygous female carriers have normal cognition and behavior, but eight showed mild physical features. All RLIM variants identified are missense changes co-segregating with the phenotype and predicted to affect protein function. Eight of the nine altered amino acids are conserved and lie either within a domain essential for binding interacting proteins or in the C-terminal RING finger catalytic domain. In vitro experiments revealed that these amino acid changes in the RLIM RING finger impaired RLIM ubiquitin ligase activity. In vivo experiments in rlim mutant zebrafish showed that wild type RLIM rescued the zebrafish rlim phenotype, whereas the patient-specific missense RLIM variants failed to rescue the phenotype and thus represent likely severe loss-of-function mutations. In summary, we identified a spectrum of RLIM missense variants causing syndromic XLID and affecting the ubiquitin ligase activity of RLIM, suggesting that enzymatic activity of RLIM is required for normal development, cognition and behavior.

Acheiropodia, congenital limb truncation, is associated with homozygous deletions in the LMBR1 gene around ZRS, an enhancer regulating SHH during limb development. How these deletions lead to this phenotype is unknown. Using whole-genome sequencing, we fine-mapped the acheiropodia-associated region to 12 kb and show that it does not function as an enhancer. CTCF and RAD21 ChIP-seq together with 4C-seq and DNA FISH identify three CTCF sites within the acheiropodia-deleted region that mediate the interaction between the ZRS and the SHH promoter. This interaction is substituted with other CTCF sites centromeric to the ZRS in the disease state. Mouse knockouts of the orthologous 12 kb sequence have no apparent abnormalities, showcasing the challenges in modelling CTCF alterations in animal models due to inherent motif differences between species. Our results show that alterations in CTCF motifs can lead to a Mendelian condition due to altered enhancer–promoter interactions., Acheiropodia is associated with homozygous deletions in the LMBR1 gene around ZRS, an enhancer regulating SHH during limb development, but how these deletions lead to this phenotype is unknown. Here the authors use whole-genome sequencing, ChIP-seq, 4C-seq and DNA FISH to show that alterations in CTCF motifs are responsible via altered enhancer–promoter interactions.

Inherited metabolic disorders (IMDs) can present with psychiatric signs that vary widely from one disease to another. This picture is further complicated by the fact that these features occur at very different illness time points, which may further delay appropriate diagnosis and treatment. In this case series of 62 children and adolescents suffering from IMDs, we clustered psychiatric signs (on the basis of the fifth edition of the Diagnostic and Statistical Manual for Mental Disorders classification) as well as impaired cognitive domains (on the basis of the Research Domain Criteriamatrix) according to their mean age of onset (5.7 ± 4 years). We observed consistent patterns of occurrence across disorders. Externalizing symptoms, sleep problems, and cross‐domain self‐regulation deficits were found to precede the IMD diagnosis. Repetitive thoughts and behaviors as well as emotional dysregulation were found to occur around the disease onset. Finally, late‐onset features included dissociative or eating disorders, together with impaired emotion knowledge. Clinicians should specifically look for the co‐occurrence of age‐specific atypical signs, such as treatment resistance or worsening with psychotropic medication in the earliest stages and symptom fluctuation, confusion, catatonia, or isolated visual hallucinations. We believe that the combined characterizations of psychiatric signs and impaired neurocognitive domains may enable the earliest detection of IMDs and the appropriate care of these particular manifestations. Key Points Psychiatric signs are common in inherited metabolic disorders (IMDs) and may occur in the same age‐range as other clinical manifestations. Three clusters of psychiatric signs and two clusters of neurocognitive domains can be defined according to their mean age of onset. Warning signs to be used in liaison psychiatry should include age‐specific cognitive impairments

Background Although COPD patients are at higher risk for aspiration when breathing spontaneously, no information is available on the risk for microaspiration in invasively ventilated COPD patients. The aim of our study was to determine the relationship between COPD and abundant microaspiration in intubated critically ill patients. Methods This was a retrospective analysis of prospectively collected data, provided by 3 randomized controlled trials on microaspiration in critically ill patients receiving invasive mechanical ventilation for more than 48 h. Abundant microaspiration was defined as the presence of pepsin and or alpha-amylase at significant levels in tracheal aspirates. In all study patients, pepsin and alpha-amylase were quantitatively measured in all tracheal aspirates collected during a 48-h period. COPD was defined using spirometry criteria. Results Among the 515 included patients, 70 (14%) had proven COPD. Pepsin and alpha-amylase were quantitatively measured in 3873 and 3764 tracheal aspirates, respectively. No significant difference was found in abundant microaspiration rate between COPD and non-COPD patients (62 of 70 patients (89%) vs 366 of 445 (82%) patients, p = 0.25). Similarly, no significant difference was found in abundant microaspiration of gastric contents (53% vs 45%, p = 0.28), oropharyngeal secretions (71% vs 71%, p = 0.99), or VAP (19% vs 22%, p = 0.65) rates between the two groups. No significant difference was found between COPD and non-COPD patients in duration of mechanical ventilation, ICU length of stay, or ICU mortality. Conclusions Our results suggest that COPD is not associated with increased risk for abundant microaspiration in intubated critically ill patients.

International audience; Originally described as a risk factor for autism, CHD8 loss-of-function variants have recently been associated with a wider spectrum of neurodevelopmental abnormalities. We further expand the CHD8-related phenotype with the description of two unrelated patients who presented with childhood-onset progressive dystonia. Whole-exome sequencing conducted in two independent laboratories revealed a CHD8 nonsense variant in one patient and a frameshift variant in the second. The patients had strongly overlapping phenotypes characterized by generalized dystonia with mild-to-moderate neurodevelopmental comorbidity. Deep brain stimulation led to clinical improvement in both cases. We suggest that CHD8 should be added to the growing list of neurodevelopmental disorder-associated genes whose mutations can also result in dystoniadominant phenotypes.

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Mutations in the cytoplasmic dynein 1 heavy chain gene (DYNC1H1) have been identified in rare neuromuscular (NMD) and neurodevelopmental (NDD) disorders such as spinal muscular atrophy with lower extremity dominance (SMALED) and autosomal dominant mental retardation syndrome 13 (MRD13). Phenotypes and genotypes of ten pediatric patients with pathogenic DYNC1H1 variants were analyzed in a multi-center study. Data mining of large-scale genomic variant databases was used to investigate domain-specific vulnerability and conservation of DYNC1H1. We identified ten patients with nine novel mutations in the DYNC1H1 gene. These patients exhibit a broad spectrum of clinical findings, suggesting an overlapping disease manifestation with intermixed phenotypes ranging from neuropathy (peripheral nervous system, PNS) to severe intellectual disability (central nervous system, CNS). Genomic profiling of healthy and patient variant datasets underlines the domain-specific effects of genetic variation in DYNC1H1, specifically on toleration towards missense variants in the linker domain. A retrospective analysis of all published mutations revealed domain-specific genotype–phenotype correlations, i.e., mutations in the dimerization domain with reductions in lower limb strength in DYNC1H1–NMD and motor domain with cerebral malformations in DYNC1H1–NDD. We highlight that the current classification into distinct disease entities does not sufficiently reflect the clinical disease manifestation that clinicians face in the diagnostic work-up of DYNC1H1-related disorders. We propose a novel clinical classification for DYNC1H1-related disorders encompassing a spectrum from DYNC1H1–NMD with an exclusive PNS phenotype to DYNC1H1–NDD with concomitant CNS involvement.

International audience; Purpose: Our study aimed to evaluate potential risk factors for the development of FDICA after suprasellar tumor resection.Materials and method: After reviewing all cases of pediatric patients who benefited from a suprasellar lesion resection in our two medical institutions, we found 6 patients with a FDICA. Surgical approach strategy (pterional or subfrontal approaches) was noted. Postoperative cranial MRI was performed in each patient 3 months after surgery and every year. When a FDICA occurred, MRI was performed 6 months after the diagnosis and 1 year later to detect any progression.Results: There were 6 males with a mean age at treatment of 11 years (6 to 15). Pterional approach was performed in these 6 patients. At the 2 institutions, we have done at least 50 pterional craniotomies for suprasellar lesion resection. No FDICA was reported after subfrontal approach in 27 consecutive pediatric patients operated on from a craniopharyngioma. The delay between the surgery and the diagnosis of the FDICA was 9 months (3 to 17 months). No symptoms related to the FDICA were recorded. The mean maximal diameter of the aneurysm was 14 mm (10 to 21). ICA bifurcation was involved in 2 cases. Asymptomatic FDICA progression was noted in 2 cases but no treatment was proposed.Conclusion: The pathogenesis of FDICA is unclear, and might involve arterial wall necrosis caused by postoperative arachnoid fibrosis which might be worsened by the pterional approach.

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International audience; SHANK2 code a scaffolding protein located at the postsynaptic membrane of glutamatergic neurons. To date, only nine patients were reported with a SHANK2-variation or microdeletion. Molecular anomalies were identified through screening of large cohorts of patients, but only poor patient clinical descriptions were available. However, this information is crucial for patient care. Here, we describe two additional unrelated patients carrying a SHANK2 de novo variant, improving the delineation of the condition. Phenotypic analysis of these 11 patients identified as major features of the condition: mild to moderate intellectual disability, speech delay, poor language skills, and autism spectrum disorders.

The occurrence of 3-methylglutaconic aciduria (3-MGA) is a well understood phenomenon in leucine oxidation and ketogenesis disorders (primary 3-MGAs). In contrast, its genesis in non-canonical (secondary) 3-MGAs, a growing-up group of disorders encompassing more than a dozen of inherited metabolic diseases, is a mystery still remaining unresolved for three decades. To puzzle out this anthologic problem of metabolism, three clues were considered: (i) the variety of disorders suggests a common cellular target at the cross-road of metabolic and signaling pathways, (ii) the response to leucine loading test only discriminative for primary but not secondary 3-MGAs suggests these latter are disorders of extramitochondrial HMG-CoA metabolism as also attested by their failure to increase 3-hydroxyisovalerate, a mitochondrial metabolite accumulating only in primary 3-MGAs, (iii) the peroxisome is an extramitochondrial site possessing its own pool and displaying metabolism of HMG-CoA, suggesting its possible involvement in producing extramitochondrial 3-methylglutaconate (3-MG). Following these clues provides a unifying common basis to non-canonical 3-MGAs: constitutive mitochondrial dysfunction induces AMPK activation which, by inhibiting early steps in cholesterol and fatty acid syntheses, pipelines cytoplasmic acetyl-CoA to peroxisomes where a rise in HMG-CoA followed by local dehydration and hydrolysis may lead to 3-MGA yield. Additional contributors are considered, notably for 3-MGAs associated with hyperammonemia, and to a lesser extent in CLPB deficiency. Metabolic and signaling itineraries followed by the proposed scenario are essentially sketched, being provided with compelling evidence from the literature coming in their support., info:eu-repo/semantics/published

International audience; Research question: Primary ovarian insufficiency (POI) is defined as the early exhaustion of ovarian function, before the age of 40 years. Its origin is genetic in 20-25% of cases. In rare cases, sequence variants of the NR5A1/SF-1 gene may result in POI, or in various disorders of gonadal development (DGD) or adrenal insufficiency.Design: This study describes the cases of two families in which the association of DGD and POI enabled a diagnosis of NR5A1 deleterious variations. Their clinical, hormonal, ultrasound and genetic characteristics are reported.Results: The mothers of the affected children were 21 and 29 years when POI was diagnosed. Each nonetheless had two spontaneous pregnancies. The children have different phenotypes and different forms of DGD. None of the affected family members had adrenal insufficiency. A new sequence variant of the NR5A1 gene was identified in one family: p.Cys283Phe (c.848G>T), and the NR5A1 sequence variant c.86G>C was found in the other family.Conclusion: Sequence variation of the NR5A1 gene is a possibility that must be considered when a woman with POI or a diminished ovarian reserve has a family member or child with DGD. If a variant is identified, genetic counselling is essential for the patient and his/her family.

Developmental pathologies of the cerebellum, in particular congenital cerebellar ataxias, although rare, are the cause of often severe motor and cognitive difficulties for which there are few methods of remediation at present. Dance is a motivating and transgenerational artistic activity that involves the synchronization of the whole body with music and/or partners. In this chapter, we will begin with a brief introduction to why dance, due to its characteristics, offers particularly interesting perspectives from a therapeutic point of view. Before discussing the use of dance in the rehabilitation of neurological pathologies, we will address the particular context of congenital cerebellar ataxias, recalling the role of the cerebellum and the consequences of its impairment on the quality of life and social integration of patients and families. We will review the current possibilities of rehabilitation of children with congenital ataxia and we will highlight the absence of remedial techniques focusing on rhythm despite the rhythmic deficits induced by cerebellar damage and the important role of temporal and sensory-motor capacities during development. Finally, we will present the preliminary results of a first study conducted in our laboratory, in collaboration with the National Reference Center for Cerebellar Disorders of the University Hospital Center of Lille, which studied the benefits on motor and cognitive functions of a two-month intervention based on dance and rhythm in children with congenital cerebellar ataxia., Les pathologies développementales du cervelet, en particulier les ataxies congénitales cérébelleuses, bien que rares, sont à l’origine de difficultés motrices et cognitives souvent sévères pour lesquelles il existe peu de méthodes de remédiation à l’heure actuelle. La danse est une activité artistique motivante et transgénérationnelle qui engage la synchronisation du corps entier avec la musique et/ou des partenaires. Dans ce chapitre, nous commencerons par exposer brièvement en introduction pourquoi la danse, de par ses caractéristiques, offre des perspectives particulièrement intéressantes sur le plan thérapeutique. Avant de discuter de l’utilisation de la danse dans la réhabilitation des pathologies neurologiques, nous aborderons le contexte particulier des ataxies congénitales cérébelleuses en rappelant le rôle du cervelet et les conséquences de son atteinte sur la qualité de vie et l’intégration sociale des patients et des familles. Nous ferons le point sur les possibilités actuelles de réhabilitation des enfants porteurs d’ataxie congénitale et nous mettrons en exergue l’absence de techniques de remédiation s’intéressant au rythme malgré les déficits rythmiques induits par l’atteinte cérébelleuse et le rôle important des capacités temporelles et sensori-motrices au cours du développement. Pour terminer, nous présenterons les résultats préliminaires d’une première étude menée dans notre laboratoire, en collaboration avec le Centre National de Référence Trouble du Cervelet du Centre Hospitalier Universitaire de Lille, ayant étudié les bénéfices sur les fonctions motrices et cognitives d’une intervention de deux mois fondée sur la danse et le rythme chez des enfants présentant une ataxie congénitale cérébelleuse.

Objective To assess whether chorioamnionitis is associated with cerebral palsy (CP) or death at 2 years' corrected age in infants born before 32 weeks of gestation after spontaneous birth. Study design EPIPAGE-2 is a national, prospective, population-based cohort study of children born preterm in France in 2011; recruitment periods varied by gestational age. This analysis includes infants born alive after preterm labor or preterm premature rupture of membranes from 240/7 to 316/7 weeks of gestation. We compared the outcomes of CP, death at 2 years' corrected age, and “CP or death at age 2” according to the presence of either clinical chorioamnionitis or histologic chorioamnionitis. All percentages were weighted by the duration of the recruitment period. Results Among 2252 infants born alive spontaneously before 32 weeks of gestation, 116 (5.2%) were exposed to clinical chorioamnionitis. Among 1470 with placental examination data available, 639 (43.5%) had histologic chorioamnionitis. In total, 346 infants died before 2 years and 1586 (83.2% of the survivors) were evaluated for CP at age 2 years. CP rates were 11.1% with and 5.0% without clinical chorioamnionitis (P = .03) and 6.1% with and 5.3% without histologic chorioamnionitis (P = .49). After adjustment for confounding factors, CP risk rose with clinical chorioamnionitis (aOR 2.13, 95% CI 1.12-4.05) but not histologic chorioamnionitis (aOR 1.21, 95% 0.75-1.93). Neither form was associated with the composite outcome “CP or death at age 2.” Conclusions Among infants very preterm born spontaneously, the risk of CP at a corrected age of 2 years was associated with exposure to clinical chorioamnionitis but not histologic chorioamnionitis.

Neurochirurgie - Sous presse. Manuscrit accepte. Disponible en ligne depuis le mercredi 3 juin 2020

Limbs development is a complex process of which mecanism is today only partially known. Embryological development abnormalities of genetic origins are rare entities. Such abnormalities can be unique or multiple, single or syndromic, sporadic or of family origins.The study of large cohorts of patients carrier of limb extremities malformations is an excellent tool that allows an identification of the genes or regulatory elements involved in their pathology and consenquently, in the development of the limb. In most of the cases, the genetic event involved is a point mutation in the genes coding transcriptionnal factor or regulatory sequence. However, variations in the number of copies are also involved.Today, new technologies of genome study, from high through put sequencing of a target genes panel to a whole exome or genome sequencing, can allow an identification of these new targets. It is thank to these technological advances that we decided to study the moleculary determinism of limbs development. To do so, we analyzed a very large cohort of 684 patients, all carriers of a limb malformation, through different genes panels, of different sizes, but also through a whole exome analysis and a pangenomic CGH array.The results of this work allowed us, in the first part, to establish a genes panel, suitable to a molecular analysis laboratory, to the bioinformatic analysis with an optimized cost, and that can identify the SNVs but also the CNVs in only one analysis.On a second part, we managed to identify 5 genes, not yet described in human pathology, which seemed to have a role in limb development. For one of these genes a promising functional analysis has started.; Le développement embryonnaire des membres est un processus complexe dont le mécanisme reste à ce jour imparfaitement connu. Ses anomalies sont des entités très hétérogènes et individuellement rares, mais touchent plus de 1/500 nouveau-nés et une proportion plus élevée de foetus. Elles représentent donc un véritable problème de santé publique, d’où l’importance d’un diagnostic précis. Il peut s’agir d’anomalies uniques ou multiples, isolées ou syndromiques, sporadiques ou familiales. L’étude de larges cohortes de patients porteurs de malformations des membres est un excellent outil qui permet d’identifier des gènes ou éléments régulateurs impliqués dans leur pathologie et par conséquent dans le développement du membre. Dans la majorité des cas, l’événement génétique responsable est une mutation ponctuelle située dans des gènes codant des facteurs de transcription ou dans des régulateurs transcriptionnels. Cependant, des variations du nombre de copies peuvent être également impliquées. Actuellement, de nouvelles technologies d’étude du génome, allant du séquençage haut débit d’un panel de gènes cibles, au séquençage de l’exome complet voire du génome, peuvent permettre d’identifier ces nouvelles cibles. C’est donc grâce à l’apport de ces avancées technologiques que nous avons souhaité étudier le déterminisme moléculaire du développement des membres. Pour ce faire nous avons analysé une très large cohorte de 684 patients, tous porteurs d’une malformation des extrémités, via différents panels de gènes, plus ou moins larges, voire via l’analyse d’exome complet ou d’une CGH pangénomique enrichie. Les résultats de ce travail nous ont permis, d’une part, d’établir un panel de gènes, adapté au laboratoire d’analyse moléculaire, dont l’analyse bioinformatique et le coût sont optimisés, permettant d’identifier les SNVs mais également les CNVs en une seule technique. D’autre part, d’identifier 5 gènes peu ou non décrits en pathologie humaine mais dont le rôle dans le développement des membres semble plus que probable et dont des analyses fonctionnelles, prometteuses, ont débuté pour l’un d’entre eux.

Limbs development is a complex process of which mecanism is today only partially known. Embryological development abnormalities of genetic origins are rare entities. Such abnormalities can be unique or multiple, single or syndromic, sporadic or of family origins.The study of large cohorts of patients carrier of limb extremities malformations is an excellent tool that allows an identification of the genes or regulatory elements involved in their pathology and consenquently, in the development of the limb. In most of the cases, the genetic event involved is a point mutation in the genes coding transcriptionnal factor or regulatory sequence. However, variations in the number of copies are also involved.Today, new technologies of genome study, from high through put sequencing of a target genes panel to a whole exome or genome sequencing, can allow an identification of these new targets. It is thank to these technological advances that we decided to study the moleculary determinism of limbs development. To do so, we analyzed a very large cohort of 684 patients, all carriers of a limb malformation, through different genes panels, of different sizes, but also through a whole exome analysis and a pangenomic CGH array.The results of this work allowed us, in the first part, to establish a genes panel, suitable to a molecular analysis laboratory, to the bioinformatic analysis with an optimized cost, and that can identify the SNVs but also the CNVs in only one analysis.On a second part, we managed to identify 5 genes, not yet described in human pathology, which seemed to have a role in limb development. For one of these genes a promising functional analysis has started., Le développement embryonnaire des membres est un processus complexe dont le mécanisme reste à ce jour imparfaitement connu. Ses anomalies sont des entités très hétérogènes et individuellement rares, mais touchent plus de 1/500 nouveau-nés et une proportion plus élevée de foetus. Elles représentent donc un véritable problème de santé publique, d’où l’importance d’un diagnostic précis. Il peut s’agir d’anomalies uniques ou multiples, isolées ou syndromiques, sporadiques ou familiales. L’étude de larges cohortes de patients porteurs de malformations des membres est un excellent outil qui permet d’identifier des gènes ou éléments régulateurs impliqués dans leur pathologie et par conséquent dans le développement du membre. Dans la majorité des cas, l’événement génétique responsable est une mutation ponctuelle située dans des gènes codant des facteurs de transcription ou dans des régulateurs transcriptionnels. Cependant, des variations du nombre de copies peuvent être également impliquées. Actuellement, de nouvelles technologies d’étude du génome, allant du séquençage haut débit d’un panel de gènes cibles, au séquençage de l’exome complet voire du génome, peuvent permettre d’identifier ces nouvelles cibles. C’est donc grâce à l’apport de ces avancées technologiques que nous avons souhaité étudier le déterminisme moléculaire du développement des membres. Pour ce faire nous avons analysé une très large cohorte de 684 patients, tous porteurs d’une malformation des extrémités, via différents panels de gènes, plus ou moins larges, voire via l’analyse d’exome complet ou d’une CGH pangénomique enrichie. Les résultats de ce travail nous ont permis, d’une part, d’établir un panel de gènes, adapté au laboratoire d’analyse moléculaire, dont l’analyse bioinformatique et le coût sont optimisés, permettant d’identifier les SNVs mais également les CNVs en une seule technique. D’autre part, d’identifier 5 gènes peu ou non décrits en pathologie humaine mais dont le rôle dans le développement des membres semble plus que probable et dont des analyses fonctionnelles, prometteuses, ont débuté pour l’un d’entre eux.

International audience; Holt-Oram syndrome (HOS) is a rare genetic disease characterized by variable radial upper limb and cardiac defects. The aim of this research was to shed light on people's subjective perceptions of their diseases, how these perceptions provide meaning, and the consequences the syndrome can have in daily life and across all life stages.Semistructured interviews with ten participants diagnosed with HOS were conducted in France and analyzed using interpretative phenomenological analysis.Participants' experiences fall under two main themes, namely, "stages of self-construction as different" and "when I am no longer the only one involved", each of which has three subthemes. From childhood onwards, symptoms monopolize the physical and psychological spheres. The feeling of being different is unavoidable until the patient can appropriate his or her condition, and by the end of adolescence, the patient generally feels that he or she has adapted to the syndrome. In adulthood, other concerns arise, such as the fear of rejection, the need to better understand the genetic issues of the condition and the desire for offspring to not experience the same life difficulties.The findings underscore the specific psychological issues associated with the syndrome at different life stages and the need for holistic genetic treatment with dedicated reference centers to improve care and further address these issues.IMPLICATIONS FOR REHABILITATIONHolt-Oram syndrome is a genetic disease characterized by abnormalities of the upper limbs and shoulder girdle and associated with a congenital heart defect.Specific issues arise at different stages of life: the physical consequences of the syndrome arise during childhood, the self-construction of pervasive difference during adolescence, the fear of being rejected as a young adult, and concerns about future parenthood and the transmission of the syndrome and the desire that one's child not be confronted with the same difficulties in adulthood.The complexity and entanglement of medical and existential issues related to HOS requires the development of multidisciplinary consultations that promote holistic care.The rarity of the syndrome and the lack of knowledge about HOS among health professionals and the general public make it necessary both to establish reference centers and to create patient associations to support patients.

To evaluate in children microdose protocol compared with low dose for lower limb alignment (LLA) measurements on biplanar radiography. Children 6 years or older were included. Height, weight and hip width were measured prior to imaging. Hip width allowed classifying children into morphotype M1 (width 35 cm) corresponding to predefined acquisition parameters (kV, mA, tube speed). Micro- and low-dose protocols were used alternately, with simultaneous acquisition of frontal and lateral radiographs. LLA measurements were performed by two independent observers (n = 526). In 15 children per morphotype, a third observer performed measurements twice (n = 180). Intraclass correlation coefficients and the dose (delivered, absorbed) were calculated. 100 girls and 160 boys (mean age = 11.7 years) were investigated: 74 M1 (mean BMI, 15.7kg/m2), 149 M2 (19.8 kg/m2) and 40 M3 (30.2 kg/m2). With microdose, inter- and intra-observer agreement was >0.90 for lengths whatever the morphotype, 0.75–0.90 (M1) and >0.90 (M2, M3) for valgus/varus and flexion/hyperextension deviations. Dose reduction reached a factor of 8.5 and 5.4 for the delivered and absorbed dose respectively. Microdose could be used for LLA measurements in children and permits a significant dose reduction. • Lower limb lengths of children can be evaluated with microdose biplanar radiography. • Valgus/varus deviations also can be evaluated with microdose biplanar radiography. • Microdose biplanar radiography significantly reduces delivered and absorbed dose in children.

Split hand/foot malformation (SHFM) or ectrodactyly is a rare congenital disorder affecting limb development characterized by clinical and genetic heterogeneity. SHFM is usually inherited as an autosomal dominant trait with incomplete penetrance. Isolated and syndromic forms are described. The extent of associated malformations is highly variable and multiple syndromes with clinical and genetic overlap have been described. We report here a 28 year-old man presenting with SHFM, sparse hair and widespread freckles. Array-CGH identified a 450 kb de novo 20p12.1 microdeletion encompassing three exons (exon 6 to 8) of MACROD2. Although MACROD2 mutations have not been associated with limb malformation until now, it is located next to KIF16B, which is involved in fibroblast growth factor receptor (FGFR) signaling. Additionally, the deletion encompassed a histone modification H3K27ac mark, known as a provider of quantitative readout of promoter and enhancer activity during human limb development. Altogether, these findings suggest that the 20p12.1 CNV is causative of SHFM in the present case through disturbance of regulatory elements functioning.

International audience; The metopic suture (MS) is one of the main sutures of the calvaria; premature closure is responsible for trigonocephaly, while persistence (metopism) is considered a normal variant. The ages of onset and completion of MS closure and prevalence of metopism in normal children are poorly documented. We studied the pattern of MS closure on 3D-CT scans of 477 children admitted for head trauma since 2012. We also studied the prevalence of trigonocephaly and the sex ratio in our clinical series of patients with all types of synostosis diagnosed during the last 4 decades. In the majority of children, MS closure started at 4 months and was complete at 9 months. The prevalence of metopism was stable after 1 year of age, at 5.1%; it was more than twice as frequent in girls (F/M ratio 2.1, non-significant). Our trigonocephaly series and the literature show a steady increase in prevalence over recent decades. During the same period, the prevalence of metopism decreased steadily. Data from comparative anatomy and paleoanthropology suggest that postnatal MS persistence in our species results from the risk of dystocia caused by the closed pelvis associated with bipedalism. The increasing incidence of trigonocephaly appears to parallel the fall in prevalence of metopism. The increasing use of cesarean section may have eliminated a potent selection factor in favor of postnatal persistence of the MS.

International audience; Medulloblastomas present generally a-specifically as a fast-evolving posterior fossa tumor. Medical literature is poor concerning clinical features of medulloblastomas and their potential significance. In the present study, we reviewed 91 pediatric observations of medulloblastomas treated in Lille between 1997 and 2017. Clinical and epidemiological variables were collected and intercorrelated. They were also compared with anatomical and pathological findings, and outcome, with the aim of defining clinical-pathological entities. We also compared the group with 32 cases of posterior fossa ependymoma and 130 cases of cerebellar astrocytoma treated during the same period. We found that in medulloblastomas, the M/F ratio was higher and diagnostic delay was shorter than in astrocytomas. Also, the mean age was older than in ependymomas. Intracranial hypertension was constant; we further observed that altered general status was common (16.5%) and correlated with a metastatic tumor. We delineated two clusters: the "nodular" cluster, which associates young age, cerebello-pontine angle tumor, herniation, desmoplastic tumor, and tumor predisposition syndrome; and the "metastatic" cluster, which associates altered status, initial metastases, hydrocephalus, and diagnostic delay. Meticulous collection of clinical data at the initial phase is integral part of the oncological evaluation, with a search for genetic and prognostic risk factors, which then permits us to define clinical-pathological entities.

International audience; More and more genetic syndromes are associated with bicoronal synostosis (BCS), making non-syndromic BCS (NSBCS) a shrinking entity. However, the numerical importance and clinical impact of syndromic BCS (SBCS) versus NSBCS have not been much studied. We retrospectively reviewed our experience with BCS over the last four decades in order to compare prevalence trends in SBCS and NSBCS. 195 patients were treated for BCS during the period 1978-2017: 104 (53.3%) were syndromic, 24 (12.3%) showed clinical and/or familial features suggesting a syndrome, although without final diagnostic confirmation, and 7 (3.5%) had associated extra-cranial malformations suggesting a syndromic context without identified genetic mutation; the remaining 61 (31.3%) were purely NSBCS. Surgery was required earlier in SBCS (21.7months, 95%CI 18.4-25.1) than in NSBCS (29.5months 95%CI 26.4-32.7). Prevalence of hydrocephalus and tonsillar herniation was significantly lower in NSBCS, and mortality concerned only SBCS. Prevalence of NSBC decreased significantly over the study period, likely because of more accurate testing, and decreased slightly over the last decade, possibly because of prenatal testing and abortion. NSBCS is now much less common than SBCS, and has a less aggressive clinical course, with lower rates of hydrocephalus, tonsillar herniation and mortality. This subgroup also deserves attention because it is likely that new discoveries are still to be made.

International audience; BACKGROUND: Ependymoma is the most frequent spinal tumor in adults but it is rather uncommon in children. The aim of the present study was to retrospectively summarize the clinical and therapeutic experience in the treatment of pediatric spinal ependymomas in France.METHODS: In the present retrospective multicenter study, data from patients aged

The gene, involved in innate immune responses to bacterial peptidoglycan, has been found to be closely associated with Crohn's Disease (CD), with an Odds Ratio ranging from 3⁻36. Families with three or more CD-affected members were related to a high frequency of gene variations, such as R702W, G908R, and 1007fs, and were reported in the EPIMAD Registry. However, some rare CD multiplex families were described without identification of common linked-to-disease variations. In order to identify new genetic variation(s) closely linked with CD, whole exome sequencing was performed on available subjects, comprising four patients in two generations affected with Crohn's disease without R702W and G908R variation and three unaffected related subjects. A rare and, not yet, reported missense variation of the gene, N1010K, was detected and co-segregated across affected patients. In silico evaluation and modelling highlighted evidence for an adverse effect of the N1010K variation with regard to CD. Moreover, cumulative characterization of N1010K and 1007fs as a compound heterozygous state in two, more severe CD family members strongly suggests that N1010K could well be a new risk factor involved in Crohn's disease genetic susceptibility. 20;4

The NOD2 gene, involved in innate immune responses to bacterial peptidoglycan, has been found to be closely associated with Crohn’s Disease (CD), with an Odds Ratio ranging from 3⁻36. Families with three or more CD-affected members were related to a high frequency of NOD2 gene variations, such as R702W, G908R, and 1007fs, and were reported in the EPIMAD Registry. However, some rare CD multiplex families were described without identification of common NOD2 linked-to-disease variations. In order to identify new genetic variation(s) closely linked with CD, whole exome sequencing was performed on available subjects, comprising four patients in two generations affected with Crohn’s disease without R702W and G908R variation and three unaffected related subjects. A rare and, not yet, reported missense variation of the NOD2 gene, N1010K, was detected and co-segregated across affected patients. In silico evaluation and modelling highlighted evidence for an adverse effect of the N1010K variation with regard to CD. Moreover, cumulative characterization of N1010K and 1007fs as a compound heterozygous state in two, more severe CD family members strongly suggests that N1010K could well be a new risk factor involved in Crohn’s disease genetic susceptibility.

ObjectiveAlveolar capillary dysplasia (ACD) is one of the causes of pulmonary hypertension. Its diagnosis is histological but new pathogenetic data have emerged. The aim of this study was to describe a French cohort of patients with ACD to improve the comprehension and the diagnosis of this pathology which is probably underdiagnosed.MethodsA retrospective observational study was conducted in French hospitals. Patients born between 2005 and 2017, whose biological samples were sent to the French genetic reference centres, were included. Clinical, histological and genetic data were retrospectively collected.ResultsWe presented a series of 21 patients. The mean of postmenstrual age at birth was 37.6 weeks. The first symptoms appeared on the median of 2.5 hours. Pulmonary hypertension was diagnosed in 20 patients out of 21. Two cases had prolonged survival (3.3 and 14 months). Histological analysis was done on lung tissue from autopsy (57.1% of cases) or from percutaneous biopsy (28.6%). FOXF1 was found abnormal in 15 patients (71.4%): 8 deletions and 7 point mutations. Two deletions were found by chromosomal microarray.ConclusionThis study is one of the largest clinically described series in literature. It seems crucial to integrate genetics early into diagnostic support. We propose a diagnostic algorithm for helping medical teams to improve diagnosis of this pathology.

International audience; Mayer-Rokitansky-Küster-Hauser syndrome (MRKH) is a rare malformative disorder, characterized by congenital aplasia of the uterus and the upper two thirds of the vagina (MIM #277000). For a majority of patients, the disorder remained without identified genetic cause. However, four recurrent microdeletions, i.e. 1q21.1-16p11.2-17q12 and 22q11.21, as well as variants in genes contained in these loci, have been identified in a small number of cases. We describe an additional patient with 2q12.1q14.1 microdeletion, showing MRKH and congenital hypothyroidism due to thyroid gland hypoplasia. The patient received a dual diagnosis with microdeletion of SHOX locus in addition to the 2q12.1q14.1 microdeletion. Literature review and database analysis has enabled us to identify 5 OMIM morbid genes: CKAP2L, IL1B, IL1RN, IL36RN and PAX8. Among these, PAX8 (Paired Box Gene 8), a transcriptional factor part of the paired-box family, plays a key role in the development of the thyroid gland, kidneys and Müllerian derivatives. We discuss here the role of PAX8 and speculate on the possible involvement of PAX8 in MRKH. In this study, we report a second case of 2q12.1q14.1 microdeletion, involving PAX8 as a gene associated with Müllerian agenesis in a MRKH I and hypothyroidism. Further studies will confirm the direct participation of PAX8 in gene target sequencing in a population of MRKH with hypothyroidism.