Made available in DSpace on 2014-06-11T19:26:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-25Bitstream added on 2014-06-13T20:53:57Z : No. of bitstreams: 1 gamba_bf_me_botib.pdf: 1144074 bytes, checksum: 38b7854a72a3108b08ac952129465214 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A síndrome de Smith-Magenis (SMS) é uma complexa anomalia caracterizada por atraso no desenvolvimento neuro-psico-motor (ADNPM), anomalias craniofaciais, esqueléticas e comportamentais. Dentre as anomalias destacamos braquicefalia, baixa estatura, distúrbio do sono, comportamentos de auto-injúria e movimentos estereotipados. SMS é causada por uma deleção intersticial do cromossomo 17(p11.2) ou mutação no gene RAI1, presente nesta região cromossômica. Sua prevalência é estimada em 1:25.000 nascidos vivos podendo chegar até 1:15.000. O diagnóstico clínico para SMS é confirmado pela técnica de citogenética molecular FISH. Neste trabalho realizamos estudo citogenético por bandamento GTG, GTG em alta resolução e FISH no total de 17 casos com suspeita clínica para SMS. Para realização de FISH foram utilizadas duas sondas comerciais da CYTOCELL ®, uma contendo o gene FU1 e outra o gene RAI1. Os resultados clínicos desta casuística comparados com a literatura internacional mostrou que embora fizemos algumas considerações, demonstramos a similaridade do diagnóstico clínico de SMS nos subgrupos SMS-del e SMS-like tanto brasileiros quanto da literatura. Os resultados citogenéticos do bandamento GTG revelaram que sete casos (7/17) apresentam deleção 17(p11.2) e o FISH confirmou a deleção tanto do gene FU1 como do gene RAI1 nos sete casos identificados. Este trabalho é de grande importância por tratar de uma afecção rara, pouco conhecida, e subdiagnosticada no Brasil. Assim destacamos que o uso de ficha clínica específica para SMS é importante para a definição da hipótese diagnóstica, 7/17 casos avaliados apresentaram deleção 17p11.2, e deleção dos genes... / Smith-Magenis syndrome (SMS) is a complex anomaly characterized by developmental delay (ADNPM), craniofacial anomalies, and behavioral disorders. Among the anomalies highlight brachycephaly, short stature, sleep disturbance, behavior of self-injury and stereotyped movements. SMS is caused by an interstitial deletion of chromosome 17 (p11.2) or mutation in the retinoic acid-induced 1 (RAI1) gene, present in this chromosomal region. Its prevalence is estimated at 1:25.000 live births and may reach up to 1:15.000. The c1inical diagnosis for SMS is confirmed by molecular cytogenetic technique of FISH. In this work we performed a cytogenetic study by GTG, GTG high resolution and FISH total of 17 cases with c1inical suspicion for SMS.To perform FISH probes were used two commercial CYTOCELL ®, one containing the FLl1 gene and another gene RAI1. The c1inical results in this series compared to the literature showed that although we made some considerations, we demonstrate the similarity of the clinical diagnosis of SMS in both sub-del SMS and SMS-like Brazilians and literature. The cytogenetic results of GTG banding revealed that seven cases (7 1 17) have deletion 17(p11.2) and FISH confirmed the deletion of both the FLl1 gene and the gene RAI1 the seven cases identified. This work is of great importance for treating arare, little known and under-diagnosed in Brazil. Thus we emphasize that the use of medical records specific to SMS is important to define the diagnosis, 7 117 cases showed stable 17p11.2 deletion, and deletion of genes FUI and RAI, no significant differences between the c1inicalfeatures of this sample group comparing it with the literature and this is about the largest sample of Brazilian SMS already described

Made available in DSpace on 2014-06-11T19:32:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-03-03Bitstream added on 2014-06-13T19:02:39Z : No. of bitstreams: 1 vieira_gh_dr_botib.pdf: 2742179 bytes, checksum: 6bb27b4e9425723a049535fafef2bcab (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A síndrome de Smith-Magenis (SMS) foi descrita , em 1986, como uma síndrome que envolvia uma mutação na região 17p em 9 pacientes. Sua prevalência esta estimada em um caso a cada 25.000 nascidos vivos. A SMS apresenta fenótipo que inclui características físicas, no desenvolvimento e comportamentais. Os sinais faciais se caracterizam por uma face larga e de forma quadrangular, braquicefalia, frontal proeminente, sinofre, fendas palpebrais alongadas para cima, ponte nasal larga, hipoplasia de face média, nariz largo e achatado, micrognatia na infância com relativa prognatia com a idade e lábio superior protruso e em „v‟ invertido. Os sinais clínicos mais importantes na SMS são comportamentais que levam a autoagressão, hiperatividade e déficit atenção. Foram estudados 31 pacientes brasileiros com suspeita diagnóstica de SMS. As análises genéticas realizadas para avaliar este grupo incluíram técnicas de citogenética molecular (FISH), aCGH, PCR quantitativa e busca por mutações na região de transcrição do gene RAI1. Os resultados demostraram que mais de 90% dos casos neste estudo tinham deficiência mental, atraso no desenvolvimento da fala e comportamento de auto-injúria. Além disso, 30% (9/30) tiveram deleção ou mutação de ponto na região 17p11.2 e RAI1 gene, sendo que 67% apresentaram uma deleção clássica (6/9), 11% tinham uma deleção atípica (1/9) e 22% (2/9) tinham uma mutação no gene RAI1. Foi possível determinar o ponto de quebra das deleções observadas e determinar os genes envolvidos. A deleção atípica descrita neste trabalho atingiu parte do gene RAI e até o momento não havia sido descrita. Além disso, duas mutações de ponto, no exon 3 do gene foram descritas. Por fim, dentro grupo estudado, foi diagnosticado um caso com síndrome da deleção 1p36, sendo possível a sugestão de um novo... / The Smith-Magenis syndrome (SMS) was described in 1986 as a syndrome involving a deletion in the 17p region in 9 patients. Its prevalence is estimated at one case per 25.000 live births. The SMS has phenotype that includes physical and behavioral development. The facial features are characterized by brachycephaly, midface hypoplasia, relative prognatism, everted, tented upper lip and deep-set, close-spaced eyes. The most important clinical features in SMS are leading behavioral self-injury, hyperactivity and attention deficit. We studied 31 Brazilian patients with suggested diagnostic to SMS. The genetic analysis performed to evaluate this group included molecular cytogenetic techniques (FISH), aCGH, quantitative PCR and the search for mutations in the gene transcription RAI1. Results showed that over 90% of the cases in this study had intellectual disability, delayed speech-language development, and self-injurious behavior. Furthermore, 30% had deletion or point mutation in the 17p11.2 region and RAI1 gene. Within this group, we found that 67% carried a classic deletion, 11% had an atypical deletion and 22% had a mutation in the RAI1 gene. It was possible to determine the breakpoint of the deletions observed and to determine the genes involved. The atypical deletion described reached part of the gene RAI1 and to date had not been described. In addition, two point mutations in exon 3 gene have been described. Finally, in this study group, one case was diagnosed with 1p36 deletion syndrome hinting of a possible new Differential Diagnosis for SMS. These findings add information for the etiology of SMS and may facilitate the development of new diagnostic tools, including FISH probes and sequencing-based screening for mutations.

Orientador: Danilo Moretti-Ferreira / Coorientador: Anand Kumar Srivastava / Banca: Angela Maria Vianna Morgante / Banca: Lucia Regina Martelli / Banca: Claudia Domingues Bonini / Resumo: A síndrome de Smith-Magenis (SMS) foi descrita , em 1986, como uma síndrome que envolvia uma mutação na região 17p em 9 pacientes. Sua prevalência esta estimada em um caso a cada 25.000 nascidos vivos. A SMS apresenta fenótipo que inclui características físicas, no desenvolvimento e comportamentais. Os sinais faciais se caracterizam por uma face larga e de forma quadrangular, braquicefalia, frontal proeminente, sinofre, fendas palpebrais alongadas para cima, ponte nasal larga, hipoplasia de face média, nariz largo e achatado, micrognatia na infância com relativa prognatia com a idade e lábio superior protruso e em „v‟ invertido. Os sinais clínicos mais importantes na SMS são comportamentais que levam a autoagressão, hiperatividade e déficit atenção. Foram estudados 31 pacientes brasileiros com suspeita diagnóstica de SMS. As análises genéticas realizadas para avaliar este grupo incluíram técnicas de citogenética molecular (FISH), aCGH, PCR quantitativa e busca por mutações na região de transcrição do gene RAI1. Os resultados demostraram que mais de 90% dos casos neste estudo tinham deficiência mental, atraso no desenvolvimento da fala e comportamento de auto-injúria. Além disso, 30% (9/30) tiveram deleção ou mutação de ponto na região 17p11.2 e RAI1 gene, sendo que 67% apresentaram uma deleção clássica (6/9), 11% tinham uma deleção atípica (1/9) e 22% (2/9) tinham uma mutação no gene RAI1. Foi possível determinar o ponto de quebra das deleções observadas e determinar os genes envolvidos. A deleção atípica descrita neste trabalho atingiu parte do gene RAI e até o momento não havia sido descrita. Além disso, duas mutações de ponto, no exon 3 do gene foram descritas. Por fim, dentro grupo estudado, foi diagnosticado um caso com síndrome da deleção 1p36, sendo possível a sugestão de um novo... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The Smith-Magenis syndrome (SMS) was described in 1986 as a syndrome involving a deletion in the 17p region in 9 patients. Its prevalence is estimated at one case per 25.000 live births. The SMS has phenotype that includes physical and behavioral development. The facial features are characterized by brachycephaly, midface hypoplasia, relative prognatism, everted, "tented" upper lip and deep-set, close-spaced eyes. The most important clinical features in SMS are leading behavioral self-injury, hyperactivity and attention deficit. We studied 31 Brazilian patients with suggested diagnostic to SMS. The genetic analysis performed to evaluate this group included molecular cytogenetic techniques (FISH), aCGH, quantitative PCR and the search for mutations in the gene transcription RAI1. Results showed that over 90% of the cases in this study had intellectual disability, delayed speech-language development, and self-injurious behavior. Furthermore, 30% had deletion or point mutation in the 17p11.2 region and RAI1 gene. Within this group, we found that 67% carried a classic deletion, 11% had an atypical deletion and 22% had a mutation in the RAI1 gene. It was possible to determine the breakpoint of the deletions observed and to determine the genes involved. The atypical deletion described reached part of the gene RAI1 and to date had not been described. In addition, two point mutations in exon 3 gene have been described. Finally, in this study group, one case was diagnosed with 1p36 deletion syndrome hinting of a possible new Differential Diagnosis for SMS. These findings add information for the etiology of SMS and may facilitate the development of new diagnostic tools, including FISH probes and sequencing-based screening for mutations. / Doutor

Orientador: Danilo Moretti-Ferreira / Banca: Rodrigo Affonseca Bressan / Banca: Antonia Paula Marques de Faria / Resumo: A síndrome de Smith-Magenis (SMS) é uma complexa anomalia caracterizada por atraso no desenvolvimento neuro-psico-motor (ADNPM), anomalias craniofaciais, esqueléticas e comportamentais. Dentre as anomalias destacamos braquicefalia, baixa estatura, distúrbio do sono, comportamentos de auto-injúria e movimentos estereotipados. SMS é causada por uma deleção intersticial do cromossomo 17(p11.2) ou mutação no gene RAI1, presente nesta região cromossômica. Sua prevalência é estimada em 1:25.000 nascidos vivos podendo chegar até 1:15.000. O diagnóstico clínico para SMS é confirmado pela técnica de citogenética molecular FISH. Neste trabalho realizamos estudo citogenético por bandamento GTG, GTG em alta resolução e FISH no total de 17 casos com suspeita clínica para SMS. Para realização de FISH foram utilizadas duas sondas comerciais da CYTOCELL ®, uma contendo o gene FU1 e outra o gene RAI1. Os resultados clínicos desta casuística comparados com a literatura internacional mostrou que embora fizemos algumas considerações, demonstramos a similaridade do diagnóstico clínico de SMS nos subgrupos SMS-del e SMS-like tanto brasileiros quanto da literatura. Os resultados citogenéticos do bandamento GTG revelaram que sete casos (7/17) apresentam deleção 17(p11.2) e o FISH confirmou a deleção tanto do gene FU1 como do gene RAI1 nos sete casos identificados. Este trabalho é de grande importância por tratar de uma afecção rara, pouco conhecida, e subdiagnosticada no Brasil. Assim destacamos que o uso de ficha clínica específica para SMS é importante para a definição da hipótese diagnóstica, 7/17 casos avaliados apresentaram deleção 17p11.2, e deleção dos genes... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Smith-Magenis syndrome (SMS) is a complex anomaly characterized by developmental delay (ADNPM), craniofacial anomalies, and behavioral disorders. Among the anomalies highlight brachycephaly, short stature, sleep disturbance, behavior of self-injury and stereotyped movements. SMS is caused by an interstitial deletion of chromosome 17 (p11.2) or mutation in the retinoic acid-induced 1 (RAI1) gene, present in this chromosomal region. Its prevalence is estimated at 1:25.000 live births and may reach up to 1:15.000. The c1inical diagnosis for SMS is confirmed by molecular cytogenetic technique of FISH. In this work we performed a cytogenetic study by GTG, GTG high resolution and FISH total of 17 cases with c1inical suspicion for SMS.To perform FISH probes were used two commercial CYTOCELL ®, one containing the FLl1 gene and another gene RAI1. The c1inical results in this series compared to the literature showed that although we made some considerations, we demonstrate the similarity of the clinical diagnosis of SMS in both sub-del SMS and SMS-like Brazilians and literature. The cytogenetic results of GTG banding revealed that seven cases (7 1 17) have deletion 17(p11.2) and FISH confirmed the deletion of both the FLl1 gene and the gene RAI1 the seven cases identified. This work is of great importance for treating arare, little known and under-diagnosed in Brazil. Thus we emphasize that the use of medical records specific to SMS is important to define the diagnosis, 7 117 cases showed stable 17p11.2 deletion, and deletion of genes FUI and RAI, no significant differences between the c1inicalfeatures of this sample group comparing it with the literature and this is about the largest sample of Brazilian SMS already described / Mestre

Smith-Magenis syndrome (SMS) is a complex mental retardation syndrome caused by deletion of 17p11.2 region or mutation of the RAI1 gene (retinoic acid induced 1). Individuals with SMS typically exhibit speech and motor delays, mental retardation, characteristic craniofacial and skeletal anomalies, and a distinct neurobehavioral phenotype that includes sleep disturbances, stereotypes, and maladaptive and self-injurious behaviors. RAI1 is thought to be a transcription factor modulating the expression of genes involved in a variety of cellular functions. Previous studies have shown the RAI1 gene being induced by retinoic acid (RA), a derivative of vitamin A. RA plays a significant role in many processes such as immune function, neurogenesis and reproduction, and deprivation of RA causes craniofacial defects. We hypothesized that RA could be inducing RAI1 which then acts as a transcription factor in modulating the expression of multiple genes. To understand the consequences of clinical variation of RAI1 gene, we performed mutation screening and identified the first case of SMS without mental retardation. Using a zebrafish model, full-length rai1 gene was cloned and spatial and temporal expression of rai1 by in-situ hybridization was evaluated and the effect of RA on rai1 expression was subsequently analyzed. The data show rai1 expression in forebrain (diencephalon) and midbrain. A rai1 antisense morpholino will eventually be created to perform knockdown studies and rescue experiments. These studies will help in determining the significance of the rai1 gene, and its interacting molecular pathways responsible for growth, development, and behavior.

Hearing loss (HL) is one of the most common sensorineural disorders and several dozen genes contribute to its pathogenesis. Establishing a genetic diagnosis of HL is of great importance for clinical evaluation of deaf patients and for estimating recurrence risks for their families. Efforts to identify genes responsible for HL have been challenged by high genetic heterogeneity and different ethnic-specific prevalence of inherited deafness. Here we present the utility of whole exome sequencing (WES) for identifying candidate causal variants for previously unexplained nonsyndromic HL of seven patients from four unrelated Altaian families (the Altai Republic, South Siberia). The WES analysis revealed homozygous missense mutations in three genes associated with HL. Mutation c.2168A>G (SLC26A4) was found in one family, a novel mutation c.1111G>C (OTOF) was revealed in another family, and mutation c.5254G>A (RAI1) was found in two families. Sanger sequencing was applied for screening of identified variants in an ethnically diverse cohort of other patients with HL (n = 116) and in Altaian controls (n = 120). Identified variants were found only in patients of Altaian ethnicity (n = 93). Several lines of evidences support the association of homozygosity for discovered variants c.5254G>A (RAI1), c.1111C>G (OTOF), and c.2168A>G (SLC26A4) with HL in Altaian patients. Local prevalence of identified variants implies possible founder effect in significant number of HL cases in indigenous population of the Altai region. Notably, this is the first reported instance of patients with RAI1 missense mutation whose HL is not accompanied by specific traits typical for Smith-Magenis syndrome. Presumed association of RAI1 gene variant c.5254G>A with isolated HL needs to be proved by further experimental studies.

Smith-Magenis Syndrome (SMS; OMIM #182290) is a multiple congenital abnormality and intellectual disability (ID) disorder caused by either an interstitial deletion of the 17p11.2 region containing the retinoic acid induced-1 (RAI1) gene or a mutation of the RAI1 gene. Individuals diagnosed with SMS typically present characteristics such as ID, self-injurious behavior, sleep disturbance, ocular and otolaryngological abnormalities, craniofacial and skeletal abnormalities, neurological and behavioral abnormalities, as well as other systemic defects and manifestations. Previous work by Vyas in 2009 showed temporal expression of rai1 in zebrafish embryos as early as 9 hpf. We hypothesize that there is maternal rai1 expression as early as zero hours post fertilization in wild type embryos. Using end-point PCR, we found that in fact there is maternal rai1 expression is detectable as early as 2 hours post fertilization (hpf) in wild type zebrafish embryos. Furthermore, we quantified rai1 expression using qPCR and found that rai1 expression declines significantly after 6 hpf. We hypothesize that a down regulation of rai1 or loss of rai1 will lead to morphological phenotypes, especially if that loss of rai1 function occurs during the earliest stages of zebrafish embryogenesis. Using a rai1morpholino oligonucleotide (MO), we found a loss of rai1 expression did not induce a morphological phenotype in in wild type embryos; furthermore, we also found that a loss of maternal rai1 expression did not induce a morphological phenotype as well. Utilizing a mutant rai1 zebrafish line, we found that both rai1 +/fh370 progeny nor rai1 fh370/fh370 progeny exhibited a morphological phenotype and that downstream targets such as bdnf were not affected by a reduction or complete loss of rai1. Prior research has shown that retinoic acid (RA) can induce rai1 expression. We hypothesize that RA can induce expression of rai1 during zebrafish embryogenesis. Using wild type fish and a rai1 in situ hybridization probe, we found that RA treatment at 25 hpf induced expression of rai1. The construction of a rai1 overexpression vector used for overexpression studies was started. Further development of GFP expression vector and zebrafish rai1 antibody are needed to determine if the morpholino is reducing rai1 protein expression.

Smith-Magenis syndrome (SMS) is a complex disorder caused by haploinsufficiency of RAI1 and characterized by sleep disturbances, behavioral abnormalities, mental retardation, and obesity in teens and adults. Rai1+/- mice are obese after 20 weeks. Dup(17)(p11.2) syndrome is a complex disorder associated with overexpression of RAI1. A transgenic mouse model of dup(17)(p11.2) syndrome overexpresses Rai1 and results in a mouse that is growth delayed. In order to characterize the obese phenotypes of mouse models of SMS and the role of RAI1 in obesity, daily food intake and serum levels of insulin, glucose, PPY, and leptin were measured; adiposity was studied by characterizing fat deposition; and gene expression was studied in the hypothalamus. These studies show that Rai1+/- mice are hyperphagic, consume more during the inactive light phase, and have altered satiety genes in the hypothalamus. Adiposity studies have shown WT females have a higher body fat content and visceral fat proportion than males, but Rai1-Tg and Rai1+/- females have similar fat deposition patterns as WT males. Hypothalamic gene expression studies show that many genes and pathways are affected by Rai1 and Rai1 dosage, including many genes associated with obesity and satiety.

Smith-Magenis syndrome (SMS) is a multiple congenital abnormalities intellectual disability syndrome that results from a deletion of chromosome 17p11.2 or mutation of the retinoic acid inducted one gene (RAI1). SMS is characterized by a multitude of phenotypic features including craniofacial defects, short stature, obesity, intellectual disability, self-abusive behavior, sleep disturbance and behavioral abnormalities. Interestingly, although SMS is a clearly defined syndrome with a known molecular change at its foundation, ~40% of all candidate cases sent to the Elsea lab for evaluation do not have a mutation or deletion of RAI1. We hypothesize that at least one other locus must be responsible for this Smith-Magenis-like (SMS-like) phenotype. To address this hypothesis, we first compiled a cohort of 52 subjects who had been referred to the Elsea lab for a clinical diagnosis of SMS. Once these individuals were confirmed to not have an RAI1 mutation or deletion, their phenotypes were compiled and statically analyzed to distinguish whether SMS and SMS-like cohorts are different in the prevalence of the core phenotypes of SMS such as, but not limited to, sleep disturbance, self-abusive behavior and developmental delay. SMS-like and SMS cohorts are not different in prevalence for these core features. Next, all SMS-like subjects were sent for whole genome array comparative genomic hybridization (aCGH) to identify duplications or deletions of each individual’s genome which contribute to the phenotype observed. We identified 6 pathogenic copy number variants (CNVs) in six individuals which contribute directly to the clinical phenotype, including two del(2)(q37). This study enabled us to draw relationships between SMS and other syndromes that had never been appreciated before and helped to identify pathways in which RAI1 may function. Using the data from our SMS-like study we were able to further characterize two known syndromes; Deletion 2q37 syndrome (brachydactyly mental retardation syndrome) and deletion 2q23 syndrome. With regard to deletion 2q37, syndrome we used genomic data from known and new deletion 2q37 subjects to refine the critical region to one gene: the histone deacetylase 4 gene (HDAC4). Using both clinical and molecular clues, we were able to identify one subject from our SMS-like cohort who has an insertion in HDAC4 which results in a premature stop codon. We conclude from this study that mutation of HDAC4 results in brachydactyly mental retardation syndrome. With regard to deletion 2q23 syndrome there were only five known cases in the published literature to which we were able to add two more. Using as similar approach to our del2q37 study we refined the critical region for this syndrome to one gene, the methyl binding domain 5 gene (MBD5). Using a molecular and clinical approach we were able to conclude that haploinsufficiency of MBD5 results in the core phenotypes seen in del2q23 syndrome including microcephaly, intellectual disabilities, severe speech impairment, and seizures. Using all the data generated from the three previous studies we set out to characterize the molecular function of RAI1. We hypothesize that RAI1 is a transcription factor that regulates gene expression of core genes involved in development, neurological function, and circadian rhythm. Using a ChIP-chip based approach we identified 257 transcripts we believe RAI1 regulates. Following up on these transcripts, using in vitro and in vivo methods, we have been able to conclude that RAI1 is a positive regulator of CLOCK, the master regulator of the central circadian cycle. Taken together, these studies have given us insight into the specific molecular changes that contribute to SMS and SMS-like syndromes. We have unveiled pathways and genes which are important to normal human development and behavior and identified novel functions of RAI1. These studies will provide the foundation for the future discovery of the pathways affected.

This diploma thesis is focused on education specifics of children with a rare disease at a special primary school. In the theoretical part, the author describes three specific rare diseases - mucopolysaccharidosis, Prader-Willi syndrome and Smith-Magenis syndrome. Deals with the manifestations of the disease, treatment options and the arrangement of the education for these children. In the practical part the author inquires how teachers work with a child with a rare disease during the day at special primary school. The thesis also inquires what specifics the educational process brings to teachers and where do the teachers find the sources for their methods of work.