Obesidade sindrômica é definida como a obesidade ocorrendo em conjunto com várias características clínicas distintas, associadas a retardo mental. A forma sindrômica mais freqüente é a síndrome de Prader-Willi (PWS) caracterizada por hipotonia, dificuldade de sucção no período neonatal, atraso do desenvolvimento neuropsicomotor (DNPM), hiperfagia, obesidade, baixa estatura na adolescência, mãos e pés pequenos, hipogonadismo, dificuldade de aprendizado e distúrbios de comportamento. Estudamos 141 pacientes com obesidade e/ou hiperfagia, atraso no desenvolvimento neuropsicomotor e/ou dificuldades de aprendizado e distúrbios de comportamento, pela técnica de MLPA (multiplex ligation-dependent probe amplification) assim como 19 pacientes que apresentavam além de atraso do DNPM e/ou dificuldade de aprendizado, distúrbios de comportamento, obesidade e/ou hiperfagia, outro sinal ao exame físico que sugerisse alteração cromossômica, pela técnica de SNP-array (The GeneChip 174; Mapping 100K Set, Affymetrix), com o objetivo de identificar genes e/ou segmentos cromossômicos envolvidos com obesidade sindrômica. Essas técnicas detectam deleções e/ ou duplicações do genoma, seja analisando regiões específicas, como a de MLPA, seja cobrindo praticamente o genoma inteiro (SNP-array). Dez pacientes apresentaram alterações cromossômicas: duas deleções 1p36, uma deleção 2p25.3, uma deleção 3p26.3 e duplicação 11q22.3, uma deleção 6(q16.1-q21), duas deleções 12(q15q21.1) (irmãs gêmeas), uma deleção X(p22.13p22.12), uma duplicação 14q11.2 e uma duplicação X(q26.3). Dentre as alterações encontradas estão duas síndromes relacionadas com obesidade já descritas, a monossomia 1p36 e a monossomia 6q16, que são diagnósticos diferencias da PWS. Nos segmentos alterados foram localizados vários genes relacionados a obesidade: DRD2, MCHR2, PLCH2, PRKCZ, RAB21, RAB2B, RAB39, TPO e SIM1. Onze genitores foram analisados por MLPA, SNP-Array e/ou cariótipo e rearranjos cromossômicos não foram identificados. Na presença dos cromossomos parentais normais o risco de recorrência é considerado desprezível. O diagnóstico de pacientes com obesidade sindrômica é um desafio, pois há sobreposição de fenótipos impossibilitando até agora o diagnóstico diferencial, a não ser o da síndrome de Prader-Willi clinicamente reconhecível, pelo menos, em sua segunda fase. O emprego de técnicas que detectam variações no número de cópias do genoma humano amplia a possibilidade de reconhecimento de novas síndromes e a descrição do espectro da variabilidade fenotípica de síndromes conhecidas. Estas síndromes são uma potencial fonte de esclarecimento das causas das formas comuns de obesidade. / Syndromic obesity is defined as obesity occurring in association with several distinct clinical features and mental retardation (MR). Prader-Willi syndrome (PWS) is the most frequent syndromic form of obesity and is characterized by hypotonia, poor sucking in the neonatal period, developmental delay, hyperphagia, obesity, short stature in adolescence, small hands and feet, hypogonadism, learning disabilities and behavior disturbances. Herein, we studied 141 patients with obesity and/or hyperphagia, psychomotor developmental delay and/or learning disabilities and behavior disturbances with the technique of MLPA (multiplex ligation-dependent probe amplification), and 19 patients by SNP-array technique (\"The GeneChip 174; Mapping 100K Set, Affymetrix) to identify copy number variations. By using both techniques we detected deletions or duplications of the genome in ten patients: two deletions at 1p36, two deletions at 12q15q21.1 (twins), a deletion of chromosomes 2p25.3, 6q16.1-q21, and Xp22.13p22.12, a duplication of chromosomes 14q11.2 and Xq26.3, and an unbalanced translocation between chromosomes 3p26.3 and 11q22.3. Monosomy 1p36 and monosomy 6q16 are well-known syndromes and had already been related with obesity. Both syndromes are considered as differential diagnosis of PWS. Several genes related to obesity are mapped in the altered chromosome segments: DRD2, MCHR2, PLCH2, PRKCZ, RAB21, RAB2B, RAB39, TPO and SIM1. Eleven parents were studied by MLPA, SNP array, and / or karyotype analyses, and chromosomal rearrangements were not identified. Therefore, we consider these rearrangements to be causative of the patients´ phenotype. The diagnosis of patients with syndromic obesity is a challenge due to the overlapping of the phenotypes, except for Prader-Willi syndrome that is a clinically recognizable syndrome, mainly in its second phase. The use of techniques that detect copy number variations of the human genome will increase the recognition of new syndromes and also the description of the spectrum of phenotypic variability of known syndromes. These syndromes are a potential source for the understanding of the etiology of the common forms of obesity.

L’épidémie actuelle d’obésité est devenue un enjeu majeur de Santé Publique dans les pays industrialisés, mais aussi dans les pays en voie de développement. Elle expose les personnes concernées à de nombreuses maladies cardio-vasculaires, métaboliques, articulaires, cancéreuses, et à une augmentation de la mortalité. Si à l’échelle sociétale, l’obésité est liée aux récents changements de notre mode de vie (accès facile à une alimentation hypercalorique, combiné à une diminution de l’activité physique), à l’échelle individuelle c’est la génétique qui détermine en grande partie notre corpulence. Dans l’obésité commune, polygénique, l’héritabilité du poids est de 70% et 5% des obésités sont monogéniques, dues à une mutation d’un seul gène dans la voie leptine-mélanocortine, régulatrice de la satiété. La diffusion des nouvelles technologies de séquençage haut débit (NGS pour Next Generation Sequencing) a permis des avancées considérables sur la connaissance de la maladie et offre une meilleure efficacité que les méthodes classiques par séquençage Sanger ou par puces à ADN, avec une plus grande rapidité de diagnostic génétique et souvent à moindre coût. Lors de mon travail de thèse, nous avons instauré une collaboration entre les services de Pédiatrie et le Centre de Génétique Chromosomique l’Hôpital Saint-Vincent de Paul à Lille d’une part, et le laboratoire UMR 8199 d’autre part afin d’appliquer ces nouvelles techniques de séquençage au diagnostic génétique d’enfants obèses suivis dans le service. Nous avons d’abord étudié 283 enfants obèses ou en surpoids adressés au Centre de Génétique Chromosomique pour bilan étiologique d’un retard de développement psychomoteur. Les premières analyses génétiques (caryotype, CGH array et recherche d’un syndrome de Prader-Willi par analyse du profil de méthylation) étaient normales et le tableau clinique n’était pas expliqué. Nous avons identifié deux nouvelles mutations délétères de SIM1 (c.886A>G/p.R296G et c.925A>G/p.S309G) chez deux patients caucasiens aux phénotypes différents. Ce travail a fait l’objet d’une publication dans le journal « Obesity ».Dans un deuxième temps, nous avons développé un protocole basé sur le NGS pour la détection simultanée des mutations ponctuelles et des anomalies structurales du génome, délétion ou duplications (CNV pour Copy Number Variation). Nous avons vérifié la capacité de notre capture personnalisée à détecter les CNV connus chez 40 patients du Centre de Génétique Chromosomique porteurs de troubles intellectuels. Puis le protocole a été appliqué chez 29 enfants obèses suivis à l’Hôpital Saint-Vincent de Paul permettant chez un certain nombre d’entre eux l’identification d’anomalies génétiques causales en une seule étape et par conséquence une amélioration de leur prise en charge médicale.Lors de mon travail de thèse, nous avons ainsi montré l’intérêt de l’utilisation des nouvelles techniques de séquençage issues du NGS pour améliorer le diagnostic génétique des enfants suspects d’obésité syndromique ou monogénique, première étape indispensable à l’émergence d’une médecine personnalisée dans la prise en charge de ces enfants. / Obesity has become a major public health issue in industrialized countries as well as in developing countries, exposing obeses patients to many cardiovascular disease, metabolic disease, osteoarthritis and cancer, and to an increasing mortality. The current worldwide obesity epidemic has largely been driven by recent changes in our lifestyle (easy access to a hypercaloric food, combined with a reduction of physical activity), however genetic differences have an appreciable role in the observed individual corpulence variation. In polygenic obesity, weight heritability is 70% and 5% of obesities are monogenic, due to a mutation of a single gene in the leptin-melanocortin pathway, regulating satiety. The use of next generation sequencing (NGS) has led to significant advances in disease awareness and provides better efficacy than conventional Sanger sequencing or DNA-chips with more accurate, faster and cheaper genetic diagnosis.During my PhD, we established a collaboration between the Pediatric departments and the Chromosomal Genetics Center at the Hôpital Saint-Vincent de Paul in Lille on the one hand and the laboratory UMR 8199 on the other hand, in order to apply the NGS to the genetic diagnosis of obese children followed in the service. We first studied 283 obese or overweight children referred to the Chromosomal Genetics Center for aetiological assessment of a developmental delay. The first genetic analyzes (karyotype, CGH array and analysis of the methylation profile for a Prader-Willi syndrome) were normal and the clinical phenotype was not explained. We identified two new deleterious mutations of SIM1 (c.886A> G / p.R296G and c.925A> G / p.S309G) in two Caucasian patients with different phenotypes. This work was published in "Obesity".We also developed an NGS-based protocol for the simultaneous detection of punctual mutations and structural anomalies of the genome, deletion or duplication (CNV for Copy Number Variation). We verified the ability of our personalized capture to detect the CNVs known in 40 patients of the Center of Chromosomal Genetics carrying intellectual disorders. The protocol was then applied to 29 obese children from Saint-Vincent de Paul Hospital, enabling a certain number of them to identify causal genetic abnormalities in a single step and, consequently, improving their medical management.In conclusion, during my PhD, we showed the advantage of using the new sequencing technologies derived from the NGS during the genetic diagnosis for children suspected of syndromic or monogenic obesity, this being the first essential step for the emergence of a personalized medicine in the management of childhood obesity

Objectifs : Documenter la prévalence de possibles retards de développement sur les domaines cognitif-langagier, moteur et socio-affectif chez les jeunes enfants négligés suivis en protection de la jeunesse (PJ) au Québec. Explorer l’association entre les possibles retards de développement et les sous-types de négligence. Méthode : Cette étude transversale a utilisé un échantillon composé de 423 enfants tirés de la banque de données clinico-administratives des services PJ du Québec. Les critères de sélection étaient les suivants : 1) être âgé entre 0 et 5 ans au moment de la rétention du signalement à l’étape « évaluation-orientation » pour motif de négligence et 2) avoir été évalué à la Grille d’évaluation du développement de l’enfant (GED) par un intervenant des services PJ. Résultats : 66,44 % des enfants ont obtenu des scores au GED les situant dans une zone à risque dans au moins un domaine du développement du jeune enfant. Ces possibles retards se déclinent ainsi : 54,6 % cognitive-langagier, 37,1 % moteur et 30,4 % socio-affectif. Des analyses bivariées et de régression logistique ont été utilisées. La négligence de soins de santé était associée à un développement socio-affectif suspect. La négligence éducative était associée à de possibles retards sur le plan cognitif-langagier et moteur. Conclusion : La prévalence de possibles retards de développement chez les enfants négligés suivis en PJ au Québec est élevée. Certains sous-types de négligence apparaissent comme étant associés à de possibles retards dans des domaines spécifiques du développement. Ces constats appuient la nécessité de dépister les retards de développement chez les jeunes enfants négligés et de leur offrir rapidement des interventions précoces et spécialisées afin de leur permettre de rattraper les jalons de développement manquants. De plus amples recherches devront être réalisées pour mieux comprendre les sous-types de négligence et les mécanismes qui sous-tendent leur relation au développement de l’enfant. / Objectives: Estimate the prevalence of suspected developmental delay among neglected children in Quebec’s child welfare system and explore the relationship with neglect subtypes. Method: In a cross-sectional approach, this study used a sample of 423 young children drawn from the anonymized database of Quebec child protection services. Selection criteria were: 1) children 0-5 years of age when report was sustained 2) for reason of neglect and 3) were assessed with a French (GED) or English (CDAS) of a child development assessment tool specifically designed for use by child welfare workers. Results: 66,44% children scored in a problem-range in at least one developmental area; 54,6% in cognitive-language, 37,1% in motor skills and 30,4% in the socioemotional area. Bivariate and logistic regression analyses were used. Medical neglect was associated with suspected developmental delay in the socioemotional area. Educational neglect was associated with suspected developmental delay in the motor and cognitive-language areas. Conclusion: Developmental concerns are high among neglected children. Neglected subtype seems related to possible delay in specific developmental areas. This finding supports early detection of developmental delay among children in the child welfare system and puts emphasis on the need of early interventions to improve developmental outcomes. Further research is needed on neglect subtypes for a better understanding of their relationship with specific developmental domains.

Secure attachment in the mother-child relationship has been shown to be predictor of positive mental health and pro-social behaviors in children who are typically developing. This study uses a sample of young children (18 mo. to 2 yrs) who had been identified as having a delay in some area of development. Mothers of these children completed two paper-pencil measures of attachment, along with measures of child temperament, maternal psychological problems, parenting stress, and child behavior problems. A second set of measures was completed one year later. Results showed that increased parenting stress and difficulty of child temperament contributed to less security of attachment, while increased maternal psychological problems predicted higher attachment security. Analysis indicated that scores on both attachment measures were stable, and that a lower degree of attachment security predicted behavior problems in this sample of children with developmental delays.

Problématique : Entre 25 et 45% des enfants nés entre 29 et 36 semaines de gestation présenteront un retard de développement (RD) à 2 ans d'âge corrigé (AC). Les enfants nés entre 29 et 36 semaines de gestation ne bénéficient d’aucun suivi systématique structuré, tel que celui retrouvé dans les Cliniques de suivi néonatal, qui permet une identification précoce des RD. Des facteurs tels que les caractéristiques périnatales, environnementales et le statut neurologique à l’âge équivalent du terme (AET) ont été étudiés comme marqueurs potentiels du RD, mais leur capacité à prédire les manifestations précoces du RD, lorsque combinés, demeure mal comprise. Objectif : Déterminer le modèle de prédiction le mieux ajusté pour la prédiction du retard moteur à 3,5 mois AC par la combinaison des facteurs périnataux, environnementaux et du statut neurologique à l’AET chez les enfants nés entre 29 et 36 semaines de gestation. Méthode: Étude prospective incluant 129 enfants nés entre 29 et 36 semaines de gestation et admis ≥48h à l’unité de soins intensifs néonatale (USIN). À l’AET, les facteurs périnataux et environnementaux ont été collectés. Un examen neurologique standardisé de l’enfant a été complété. À 3,5 mois AC, une évaluation motrice a été réalisée avec le Alberta Infant Motor Scale (AIMS): le retard était défini comme un score <10e percentile. Des régressions logistiques ont permis d’identifier les facteurs associés au RD à 3,5 mois AC et de créer un modèle de prédiction. Résultats : Les facteurs qui sont demeurés significatifs dans le modèle multivarié à 3,5 mois AC sont : consommation de tabac par la mère, présence de diabète gestationnel, score à l’Échelle de dépression postnatal d’Édimbourg, indice de risque néonatal et indice de risque environnemental. Le modèle final avait une sensibilité de 76% et une spécificité de 84% pour la prédiction du RD à 3,5 mois. Conclusion : Le modèle statistique élaboré permet une identification suffisamment sensible et spécifique pour être utilisé en clinique à des fins d’identification précoce du risque de RD auprès des enfants nés entre 29 et 36 semaines de gestation et ce, dès l’AET. / Background: Between 25 and 45% of children born prematurely between 29 and 36 weeks of gestation will present with developmental delay by 2 years of corrected age (CA). Children born between 29 and 36 weeks of gestation do not systematically benefit from specialized follow-up, which allows the early identification of developmental delay. Factors such as perinatal characteristics, environmental characteristics and neurological status have been studied as potential markers of developmental delay. However, their capacity for predicting an early manifestation of developmental delay remains poorly understood. Aims: To determine the best-fitting model for the prediction of motor delay at 3.5 months CA by the combination of perinatal factors, environmental factors, and neurological status at term equivalent age (TEA) in children born between 29 and 36 weeks' gestation. Methods: This prospective observational cohort study included 129 infants born between 29 and 36 weeks of gestation and admitted for >48h in the Neonatal Intensive Care Unit (NICU). At TEA, perinatal and environmental factors were collected, and a standardized neurological examination of the child was also performed. At 3.5 months CA, a motor assessment was performed using the Alberta Infant Motor Scale: delay was defined as a score <10th percentile. Logistic regressions were used in order to identify factors associated with motor delay at 3.5 months CA and to create a prediction model. Results: Factors that remained significant in the multivariate model at 3.5 months CA were maternal smoking, gestational diabetes, Edinburgh Postnatal Depression Scale score, neonatal risk index, and environmental risk index. The final model had a sensitivity of 76% and a specificity of 84% for predicting motor delay at 3.5 months. Conclusion: The statistical model developed demonstrated a sensitivity and specificity strong enough to support its use in a clinical context for the early identification of the risk of motor delay in children born between 29 and 36 weeks of gestation.

Introduction: A population especially overlooked in early childhood development research is the ‘missing-middle’, predominantly represented by middle-income, urban populations. Research typically focuses on populations that are either from lower- or upper socioeconomic classes. Insight into the risks, protective factors, and developmental outcomes of children from middle-income populations is needed to guide the implementation and customisation of early intervention policies and services. Aim: The study aimed to describe the developmental outcomes of young children aged six to 24 months from an urban, middle-income setting in South Africa using a descriptive correlational research design. Method: The Vineland-3 was used to describe the developmental outcomes of 55 children between six to 24 months, from an urban, middle-income setting in South Africa. The mHealth PEDS was used to identify caregivers’ concerns regarding their children’s development. Results: Caregivers’ concerns were positively associated (φ = 0.355; p = .024) with their children’s overall developmental outcomes, emphasising their valuable contribution in early identification of developmental delays. Fifteen percent (n = 8) of participants’ children presented with developmental delays, with low birth weight identified as a significant developmental risk (p = .011). Within this sample population, higher maternal education was identified as a significant protective factor (p = 0.16). Conclusion: Developmental delays, risks, strengths and protective factors exist within an urban, middle-income South African setting. Early childhood development can be promoted in all income settings by using baby wellness clinics as a point of access to identify children at risk for developmental delays through caregiver-led developmental screening. Prioritising early childhood development across different socioeconomic classes will help ensure that all children reach their full developmental potential. / Dissertation (MA (Speech-Language Pathology))--University of Pretoria, 2021. / National Research Foundation (NRF) / Speech-Language Pathology and Audiology / MA (Speech-Language Pathology) / Unrestricted

Thesis (MEdPsych)--Stellenbosch University, 2012. / Includes bibiliography / ENGLISH ABSTRACT: At birth every human being is at the starting point of many different journeys: journeys of discovery and change, and journeys of mental and physical growth. Most children follow a similar path of physical and mental growth to adulthood, achieving predetermined milestones at approximately the same age. But what happens to a child who cannot follow this path, a child born without a map? How do the diagnosis and subsequent educational journey of the child affect the parents of that child, parents who find themselves disabled by their experiences of parenthood and life? This study is an autoethnography. It was undertaken to reflect on the physical and emotional journey two parents experienced in finding an educational niche for their son who was diagnosed with Pervasive Developmental Delay – Not Otherwise Specified (PDD-NOS), an Autism Spectrum Disorder. In an attempt to engage and involve the non-academic audience, while at the same time addressing the analytical needs of the researcher audience, evocative autoethnographic co-constructed narratives were combined with analytic autoethnography. Not only did I aim to fill in some of the gaps in researcher knowledge about South African parents’ experiences in finding educational support for their children with pervasive developmental delays, but I also wanted to provide knowledge, hope and encouragement to other parents, especially those parents who are at the start of a journey leading to a brighter future for their child with special needs. / AFRIKAANSE OPSOMMING: By geboorte bevind elke mens hom by die beginpunt van verskeie reise: reise van ontdekking en verandering, en reise van geestelike en fisieke groei. Die meeste kinders volg ‘n gelyksoortige roete van geestelike en fisieke groei na volwassenheid, deur voorafbepaalde doelwitte op naastenby ooreenstemmende ouderdomme te bereik. Maar wat gebeur met ‘n kind wat nie hierdie pad kan volg nie, ‘n kind wat sonder ‘n roetekaart gebore word? Hoe beïnvloed die diagnose en gevolglike opvoedkundige reis van daardie kind sy of haar ouers, ouers wat hulself gestremd bevind in hul ervaring van ouerskap en die lewe? Hierdie studie is ‘n outo-etnografie. Dit reflekteer op die fisieke en emosionele reis deur twee ouers onderneem, in hul soeke na ‘n geskikte onderwysnis vir hul seun wat met PDD-NOS1, ‘n Outisme Spektrumversteuring, gediagnoseer is. In ‘n poging om die nie-akademiese gehoor te betrek, maar terselfdertyd die analitiese behoeftes van die navorsergehoor aan te spreek, is die tegnieke van stemmingsvolle outo-etnografiese mede-saamgestelde narratiewe en analitiese outo-etnografie gekombineer. Ek het nie slegs ten doel gehad om sommige gapings in navorsing rondom die ervarings van Suid-Afrikaanse ouers van kinders met Outisme Spektrumversteurings te vul nie, maar ook om kennis, hoop en aanmoediging te gee aan ander ouers, veral daardie ouers wat aan die begin staan van ‘n reis na ‘n beter toekoms vir hul kind met spesiale behoeftes.

A obesidade se tornou uma das maiores preocupações de saúde pública. É um distúrbio neuroendócrino, no qual fatores ambientais e genéticos agem em conjunto, levando ao excesso de armazenamento de energia na forma de gordura corporal. A síndrome de Prader-Willi (PWS) é a mais freqüente das síndromes que possui a obesidade como uma de suas características, com incidência de 1:25.000 nascimentos. É caracterizada por hipotonia neonatal com dificuldade de sucção, atraso do desenvolvimento neuropsicomotor (DNPM), hiperfagia, obesidade, baixa estatura em adolescentes, mãos e pés pequenos, hipogonadismo, distúrbios do sono, características faciais dismórficas, deficiência intelectual leve a moderada e comportamento obsessivo-compulsivo. Pacientes com atraso do DNPM e/ou dificuldade de aprendizado, distúrbios de comportamento, obesidade e/ou hiperfagia, com teste negativo para PWS, foram estudados com plataformas de SNP array, &ldquo;The GeneChip® Mapping 500K Set&rdquo; da Affymetrix, ou array-CGH, CytoSure ISCA 4x180k da OGT, para identificar genes relacionados a obesidade e hiperfagia, assim como, novas regiões genômicas implicadas na etiologia de síndromes genéticas associadas à obesidade. Dentre os 31 pacientes estudados, oito apresentaram variações de número de cópias (CNVs) em seu genoma: deleção em 1p22.1p21.2; deleção em 3q25.33q26.1 e deleção em 13q31.2q32.1; duplicação em 7q36.2; deleção em 8p23.3p23.1 e duplicação em 12p13.33p13.31; duplicação 16p13.11p12.3; duplicação em 17q11.2; deleção em 20p12.1; duplicação em 21q22.13. Duas dessas alterações foram herdadas de pais fenotipicamente normais. Algumas dessas CNVs sobrepõem regiões genômicas previamente relacionadas com obesidade, incluindo a microdeleção de 1p21.3 e as duplicações dos cromossomos 12 e 21. Identificamos genes anteriormente descritos como associados à obesidade (PTBP2, DPYD, MIR137, GNB3 e PPM1L), ou possivelmente envolvidos com este fenótipo (HTR5A e KCNJ6), mapeados em várias dessas CNVs. Além disso, os genes RNF135, NF1, DPP6, GPC5, DYRK1A e MACROD2 são os prováveis causadores da deficiência intelectual, atraso do desenvolvimento neuropsicomotor, dificuldades de aprendizagem, distúrbios de comportamento e outras características clínicas encontrados nos pacientes. O diagnóstico e prognóstico dos pacientes e o Aconselhamento Genético aos pais e familiares é fornecido / Obesity has become a major concern for public health. It is a neuroendocrine disorder, in which genetic and environmental factors act together, leading to excessive storage of energy as fat. Prader-Willi syndrome (PWS) is the main obesity-related syndrome with a birth incidence of 1:25,000. It is characterized by neonatal hypotonia, poor sucking, developmental delay, hyperphagia, obesity, short stature in adolescents, small hands and feet, hypogonadism, sleep disturbance, dysmorphic facial features, mild to moderate intellectual disability and obsessive-compulsive behavior. Patients with psychomotor developmental delay and/or learning disabilities, behavior disorders, obesity and/or hyperphagia, who tested negative for PWS, were studied by chromosomal microarray analysis, including the SNP-based platform &ldquo;The GeneChip® Mapping 500K Set&rdquo; (Affymetrix), and the array-CGH platform &ldquo;CytoSure ISCA 4x180k (OGT)&rdquo;, to identify genes related to hyperphagia and obesity, as well as new genomic regions implicated in the etiology of genetic syndromes associated with obesity. Of 31 patients studied, eight had copy number variants (CNVs) in the genome: 1p22.1p21.2 deletion; 3q25.33q26.1 deletion and 13q31.2q32.1 deletion; 7q36.2 duplication; 8p23.3p23.1 deletion and 12p13.33p13.31 duplication; 16p13.11p12.3 duplication; 17q11.2 duplicaton; 20p12.1 deletion; 21q22.13 duplication. Two of these CNVs were inherited from an unaffected father. Some of these CNVs overlap genomic regions that have previously been related to obesity, including the 1p21.3 microdeletion and the duplications of chromosomes 12 and 21. Furthermore, we identified genes previously described as associated with obesity (PTBP2, DPYD, MIR137, GNB3 and PPM1L), or possibly involved with this phenotype (HTR5A and KCNJ6), mapped to several of these CNVs. In addition, the genes RNF135, NF1, DPP6, GPC5, DYRK1A and MACROD2 are likely implicated in intellectual disability, developmental delay, learning disabilities, behavioral disorders and other clinical features found in patients. The diagnosis and prognosis of patients and genetic counseling to parents and families is provided

A obesidade se tornou uma das maiores preocupações de saúde pública. É um distúrbio neuroendócrino, no qual fatores ambientais e genéticos agem em conjunto, levando ao excesso de armazenamento de energia na forma de gordura corporal. A síndrome de Prader-Willi (PWS) é a mais freqüente das síndromes que possui a obesidade como uma de suas características, com incidência de 1:25.000 nascimentos. É caracterizada por hipotonia neonatal com dificuldade de sucção, atraso do desenvolvimento neuropsicomotor (DNPM), hiperfagia, obesidade, baixa estatura em adolescentes, mãos e pés pequenos, hipogonadismo, distúrbios do sono, características faciais dismórficas, deficiência intelectual leve a moderada e comportamento obsessivo-compulsivo. Pacientes com atraso do DNPM e/ou dificuldade de aprendizado, distúrbios de comportamento, obesidade e/ou hiperfagia, com teste negativo para PWS, foram estudados com plataformas de SNP array, &ldquo;The GeneChip® Mapping 500K Set&rdquo; da Affymetrix, ou array-CGH, CytoSure ISCA 4x180k da OGT, para identificar genes relacionados a obesidade e hiperfagia, assim como, novas regiões genômicas implicadas na etiologia de síndromes genéticas associadas à obesidade. Dentre os 31 pacientes estudados, oito apresentaram variações de número de cópias (CNVs) em seu genoma: deleção em 1p22.1p21.2; deleção em 3q25.33q26.1 e deleção em 13q31.2q32.1; duplicação em 7q36.2; deleção em 8p23.3p23.1 e duplicação em 12p13.33p13.31; duplicação 16p13.11p12.3; duplicação em 17q11.2; deleção em 20p12.1; duplicação em 21q22.13. Duas dessas alterações foram herdadas de pais fenotipicamente normais. Algumas dessas CNVs sobrepõem regiões genômicas previamente relacionadas com obesidade, incluindo a microdeleção de 1p21.3 e as duplicações dos cromossomos 12 e 21. Identificamos genes anteriormente descritos como associados à obesidade (PTBP2, DPYD, MIR137, GNB3 e PPM1L), ou possivelmente envolvidos com este fenótipo (HTR5A e KCNJ6), mapeados em várias dessas CNVs. Além disso, os genes RNF135, NF1, DPP6, GPC5, DYRK1A e MACROD2 são os prováveis causadores da deficiência intelectual, atraso do desenvolvimento neuropsicomotor, dificuldades de aprendizagem, distúrbios de comportamento e outras características clínicas encontrados nos pacientes. O diagnóstico e prognóstico dos pacientes e o Aconselhamento Genético aos pais e familiares é fornecido / Obesity has become a major concern for public health. It is a neuroendocrine disorder, in which genetic and environmental factors act together, leading to excessive storage of energy as fat. Prader-Willi syndrome (PWS) is the main obesity-related syndrome with a birth incidence of 1:25,000. It is characterized by neonatal hypotonia, poor sucking, developmental delay, hyperphagia, obesity, short stature in adolescents, small hands and feet, hypogonadism, sleep disturbance, dysmorphic facial features, mild to moderate intellectual disability and obsessive-compulsive behavior. Patients with psychomotor developmental delay and/or learning disabilities, behavior disorders, obesity and/or hyperphagia, who tested negative for PWS, were studied by chromosomal microarray analysis, including the SNP-based platform &ldquo;The GeneChip® Mapping 500K Set&rdquo; (Affymetrix), and the array-CGH platform &ldquo;CytoSure ISCA 4x180k (OGT)&rdquo;, to identify genes related to hyperphagia and obesity, as well as new genomic regions implicated in the etiology of genetic syndromes associated with obesity. Of 31 patients studied, eight had copy number variants (CNVs) in the genome: 1p22.1p21.2 deletion; 3q25.33q26.1 deletion and 13q31.2q32.1 deletion; 7q36.2 duplication; 8p23.3p23.1 deletion and 12p13.33p13.31 duplication; 16p13.11p12.3 duplication; 17q11.2 duplicaton; 20p12.1 deletion; 21q22.13 duplication. Two of these CNVs were inherited from an unaffected father. Some of these CNVs overlap genomic regions that have previously been related to obesity, including the 1p21.3 microdeletion and the duplications of chromosomes 12 and 21. Furthermore, we identified genes previously described as associated with obesity (PTBP2, DPYD, MIR137, GNB3 and PPM1L), or possibly involved with this phenotype (HTR5A and KCNJ6), mapped to several of these CNVs. In addition, the genes RNF135, NF1, DPP6, GPC5, DYRK1A and MACROD2 are likely implicated in intellectual disability, developmental delay, learning disabilities, behavioral disorders and other clinical features found in patients. The diagnosis and prognosis of patients and genetic counseling to parents and families is provided

O genoma humano é composto por diversos tipos de variações estruturais, como por exemplo, as variações no número de cópias (CNVs), que, mesmo sendo muito pequenas, podem gerar diversas alterações clínicas específicas, como as malformações congênitas e o atraso do desenvolvimento neuropsicomotor (MC/ADNPM). Para a detecção destas alterações existem diferentes técnicas citogenômicas dentre elas a FISH (Fluorescent in situ Hibridization) e a MLPA (Multiplex Ligation-dependent Probe Amplification), que investigam um número limitado de regiões do genoma, como as regiões envolvidas nas síndromes de microdeleções/microduplicações mais comuns e as regiões subteloméricas. Outros métodos como a cariotipagem clássica e o array genômico possibilitam uma análise completa do DNA em uma única reação, aumentando a taxa de detecção de desequilíbrios complexos. Alcançar um diagnóstico inequívoco é fundamental para entender a natureza da doença, fornecendo respostas sobre o prognóstico, sobre os riscos de recorrência e direcionando o paciente à terapia específica, o que pode minimizar o custo financeiro dessas doenças e até mesmo possibilitar a inclusão desses indivíduos na sociedade. O projeto teve como objetivo comparar a capacidade diagnóstica destas tecnologias (FISH, MLPA e array) para a elucidação etiológica de pacientes sindrômicos encaminhados para a unidade de genética. A casuística deste trabalho foi composta pela análise dos resultados das técnicas de FISH e/ou MLPA e array, utilizadas no diagnóstico de 78 pacientes com MC/ADNPM. Na técnica de FISH, empregada na análise genômica de 22 pacientes, foram utilizadas sondas locus específicas para as regiões das principais síndromes de microdeleção/microduplicação e para as regiões subteloméricas de cromossomos específicos. Por meio desta metodologia, foram identificados ~18,2% dos pacientes com diferentes alterações. Já a técnica de MLPA, utilizada no diagnóstico dos 78 pacientes, por meio dos kits para as principais síndromes de microdeleção/microduplicação e para as regiões subteloméricas, detectou ~34,6% de pacientes com diversas alterações. A técnica de array, realizada em todos os pacientes utilizando diferentes plataformas (Agilent, Affymetrix ou Illumina) apresentou uma taxa de ~42,3% de detecção de pacientes com pelo menos uma alteração patogênica e ~38,5% de pacientes com alterações benignas ou de significado clínico incerto. Ao avaliar as três técnicas concomitantemente foi verificada uma taxa de ~93,6% de concordância, apesar dos resultados não serem iguais em todos os casos e da técnica de MLPA não detectar ~66,2% das alterações em relação ao array. Os resultados obtidos corroboraram com dados da literatura, mas no geral a taxa de detecção foi superior às taxas descritas, o em que em parte pode ser devido ao critério de seleção dos pacientes, sugerindo fortemente que a hipótese clínica adequada é crucial para o sucesso da detecção de alteração. Embora o array seja a ferramenta mais eficiente para o diagnóstico de pacientes com malformações, seu uso como primeiro teste diagnóstico nem sempre é o mais apropriado devido ao seu custo elevado ou sua limitação em detectar inversões e translocações balanceadas. Portanto todas as técnicas estudadas têm suas vantagens e desvantagens, e poderão ser aplicadas em conjunto para que o diagnóstico molecular seja concluído. Dessa forma, são necessárias uma interação clínico-laboratorial e uma equipe técnica multiprofissional especializada para o direcionamento do diagnóstico molecular mais eficaz em relação ao custo-benefício / The human genome is composed of several types of structural variations, such as copy number variation (CNVs) which, although very small, can generate several specific clinical abnormalities, such as congenital malformations and developmental delay (CM/DD). To detect these changes there are different cytogenomics techniques, among them, FISH (Fluorescent in situ Hybridization) and MLPA (Multiplex Ligation-dependent Probe Amplification) that can investigate a limited number of genomic regions for example the most common microdeletion/microduplications syndromes and subtelomeric regions. Other methods such as classical karyotyping and array provide a complete DNA analysis in a single reaction, increasing the detection rate of complex imbalances. Acquire an unequivocal diagnosis is critical to understand the nature of the disease, providing answers about the prognosis, risks of recurrence and directing the patients to specific therapy, which can minimize the cost of these diseases and even allow the inclusion of these individuals in society. The objective of this project was to compare the diagnostic ability of these technologies (FISH, MLPA and array) for the etiologic diagnosis of syndromic patients referred to the clinical unit of genetics. The casuistry was composed by the results of analysis of 78 patients with CM/DD using FISH and/or MLPA and array. The FISH technique was utilized in genomic analysis for 22 patients and locus specific probes were used for regions of the microdeletion/microduplication syndromes and the subtelomeric regions of specific chromosomes. By this methodology ~18.2% of the patients were identified with different genomic changes. The MLPA technique was used in the diagnosis of 78 patients, with microdeletion/microduplication syndrome and subtelomeric regions, and detected ~34.6% of patients with several changes. The array technique was performed in all patients using different platforms (Agilent, Illumina or Affymetrix) and shows a rate of ~42.3% of detection at least one pathogenic change and ~38.5% of patients with benign or uncertain clinical significance changes. In assessment of the three techniques concomitantly was observed a rate of ~93.6% of concordance, although the results are not the same in all cases and the MLPA technique to detect ~ 66.2% of the changes in relation to the array. The results obtained corroborated with literature data, but the overall detection rate was higher than the rates described in the literature, due in part to the criteria selection of patients. Our results strongly suggesting that appropriate clinical hypothesis is crucial for successful change detection. Although the array is the most efficient tool for the diagnosis of patients with abnormalities, using this test as a first diagnostic approach is not always the most suitable tool because of the high cost or the limitation to detect inversions and balanced translocations. Therefore, all techniques studied have their advantages and disadvantages, and could be applied together for the completed molecular diagnosis. Thus, a clinical laboratory interaction and multidisciplinary skilled technicians is required for targeting the most effective molecular diagnosis in relation to cost-benefit