Suplemento - Revista Nascer e Crescer

Transcrição

Resumos das Comunicações
2015
XLIV Conferências de Genética Doutor Jacinto Magalhães
Suplemento I
Revista de Pediatria do Centro Hospitalar do Porto
Revista de Pediatria do Centro Hospitalar do Porto | Departamento de Ensino, Formação e Investigação
Ano | 2015 Volume | XXIV Número | Suplemento |
Diretora | Editor-in-Chief | Sílvia Álvares; Diretor Adjunto | Associated Editor | Rui Chorão; Diretora Executiva | Executive Editor | Luísa Lobato
Presidente do Conselho de Administração do Centro Hospitalar do Porto | Director | Fernando Sollari Allegro
Corpo Redatorial | Editorial Board
Conselho Científico | Scientific Board
Assessores Editoriais | Editorial Assistants
Ana Rita Araújo, ULSAM
Armando Pinto, IPOPFG
Artur Alegria, CHP
Braga da Cunha, CHTS
Carmen Carvalho, CHP
Cláudia Pedrosa, CHVNG/E
Cláudia Tavares, CHAA
Conceição Mota, CHP
Cristina Rocha, CHEDV
Fatima Carvalho, CHP
Filipa Balona, HB
Gustavo Rocha, CHSJ
Helena Sá Couto, HPH/ULSM
João Barreira, CHSJ
Laura Marques, CHP
Margarida Guedes, CHP
Maria Sameiro Faria, CHP
Vasco Lavrador, CHP
Alberto Caldas Afonso, CHSJ, FMUP, Porto
Almerinda Pereira, HB, Braga
António Tomé, CHP, Porto
Goreti Lobarinhas, HSSM Barcelos
Ana Ramos, CHP, Porto
António Martins da Silva, CHP e ICBAS/UP, Porto
Braga da Cunha, CHTS, Penafiel
Cidade Rodrigues, CHP, Porto
Cleonice Mota, FM - UFMG, Belo Horizonte
Conceição Casanova, CHPVVC, Póvoa de Varzim
Eurico Gaspar, CHTMAD, Vila Real
Fátima Praça, CHVNG/E, Vila Nova de Gaia
Gonçalves Oliveira, CHMA, Famalicão
Helena Jardim, CHP, Porto
Jorge Braga, CHP, Porto
Hercília Guimarães, CHSJ, FMUP, Porto
Herculano Rocha, CHP, Porto
Ines Lopes, CHVNG/E, Vila Nova de Gaia
José Barbot, CHP, Porto
José Carlos Areias, FMUP, Porto
José Cidrais Rodrigues, HPN/ULSM, Matosinhos
Lopes dos Santos, HPH/ULSM, Matosinhos
Luís Almeida Santos, CHSJ, FMUP, Porto
Manuel Salgado, HPCM/CHUC, Coimbra
Manuela Selores, CHP, Porto
Marcelo Fonseca, ULSM, Matosinhos
Margarida Lima, CHP, ICBAS/UP, Porto
Maria Augusta Areias, HPBN, Porto
Norberto Estevinho, HPP, Porto
Óscar Vaz, ULSN, Mirandela
Paula Cristina Ferreira, CHP, Porto
Paula Soares, CHP, Porto
Pedro Freitas, CHAA, Guimarães
Rei Amorim, CHAM, Viana do Castelo
Ricardo Costa, CHCB, Covilhã
Rosa Amorim, CHP, Porto
Rui Carrapato, CHEDV, Santa Maria da Feira
Teresa Temudo, CHP, Porto
Paulo Pinto
Editores especializados | Section Editors
Artigo Recomendado – Helena Mansilha, CHP; Maria do
Carmo Santos, CHP
Perspetivas Atuais em Bioética – Natália Teles, CGMJM
Pediatria Baseada na Evidência – Luís Filipe Azevedo,
FMUP; Altamiro da Costa Pereira, FMUP
A Cardiologia Pediátrica na Prática Clínica – António
Marinho, CHUC; Fátima Pinto, HSM/CHLC; Maria Ana
Sampaio, HCV, Maria João Baptista, CHSJ; Paula Martins,
HPCM/CHUC, Rui Anjos, HSC/CHLO; Sílvia Álvares, CHP
Ciclo de Pediatria Inter-Hospitalar do Norte – Armando
Pinto, IPOPFG; Carla Moreira, HB; Conceição Santos Silva,
CHPVVC; Fátima Santos, CHVNG/E; Inês Azevedo, CHSJ;
Isalita Moura, HSMM; Isolina Aguiar, CHAA; Joaquim Cunha,
CHTS; Laura Soares, CHEDV; Susana Tavares, CHEDV;
Cármen Carvalho, CHP; Rosa Lima, CHP; Sofia Aroso, HPH/
ULSM; Sónia Carvalho, CHMA
Caso Dermatológico – Manuela Selores, CHP; Susana
Machado, CHP
Caso Eletroencefalográfico – Rui Chorão, CHP
Caso Estomatológico – José Amorim, CHP
Caso Radiológico – Filipe Macedo, H CUF
Genes, Crianças e Pediatras – Esmeralda Martins, CHP;
Gabriela Soares, CGMJM
Educação Científica – Margarida Lima, CHP, ICBAS-UP
Pequenas Histórias – Margarida Guedes, CHP
Consultora de Epidemiologia e de Bioestatistica |
| Advisor of Epidemiology and Biostatistics
Maria José Bento, IPOPF
Paulo Silva
Publicação trimestral resumida e indexada por
Catálogo LATINDEX
EMBASE / Excerpta Médica
Index das Revistas Médicas Portuguesas
Scopus
Artigos disponíveis no Repositório Científico do CHP
http://repositorio.chporto.pt
Design gráfico
bmais comunicação
Execução gráfica e paginação
Papelmunde, SMG, Lda
Vila Nova de Famalicão
ISSN
0872-0754
Depósito legal
4346/91
Propriedade, Edição e Administração / Publisher
Departamento de Ensino, Formação e Investigação
Centro Hospitalar do Porto
Largo do Prof. Abel Salazar – 4099-001 Porto
Telefone: (+351) 222 077 500; fax: (+351) 222 082 166
Telemóvel: (+351) 915 676 516
[email protected]
CGMJM, Centro de Genética Médica Dr. Jacinto Magalhães, CHAA, Centro Hospitalar do Alto Ave; CHAM, Centro Hospitalar do Alto Minho; CHCB, Centro Hospitalar da Cova da Beira; CHEDV, Centro
Hospitalar de Entre Douro e Vouga; CHMA, Centro Hospitalar do Médio Ave; CHP, Centro Hospitalar do Porto; CHPVVC, Centro Hospitalar da Póvoa de Varzim – Vila do Conde; CHSJ, Centro Hospitalar
de São João; CHTMAD, Centro Hospitalar de Trás-os-Montes e Alto Douro; CHTS, Centro Hospitalar do Tâmega e Sousa; CHUC, Centro Hospitalar e Universitário de Coimbra; CHVNG/E, Centro
Hospitalar de Vila Nova de Gaia/Espinho; DEFI, Departamento de Ensino, Formação e Investigação;
FMUP, Faculdade de Medicina da Universidade do Porto; HB, Hospital de Braga; HCUF, Hospital da CUF,HCV, Hospital Cruz Vermelha; HPBN, Hospital Privado da Boa Nova; HPCM/CHUC, Hospital Pediátrico Carmona da Mota; HPH/ULSM, Hospital
Pedro Hispano/Unidade Local de Saúde Matosinhos; HPP, Hospitais Privados de Portugal; HSC/CHLO, Hospital de Santa Cruz/Centro Hospitalar de Lisboa Ocidental;
HSM/CHLC, Hospital de Santa Marta/Centro Hospitalar de Lisboa Central; HSMM, Hospital Santa Maria Maior; ICBAS/UP, Instituto de Ciências Biomédicas Abel Salazar da Universidade do Porto;
IPOPFG, Instituto Português de Oncologia do Porto, Francisco Gentil; ULSN, Unidade Local de Saúde do Nordeste, FM-UFMG – Faculdade de Medicina, Universidade Federal de Minas Gerais.
Resumo das Comunicações
“A Doença Rara no Adulto” - Porto, 20 Fevereiro 2015
Organização do Congresso l Congress Organization
Cristina Candeias
Dulce Quelhas
Francisco Laranjeira
Isabel Marques
Márcia Oliveira
Natália Oliva Teles
Comissão Científica l Scientific Committee
Ana Fortuna
Lúcia Lacerda
Natália Oliva Teles
Paula Jorge
Colaboração logística das Conferências l Local Support
CGMJM/CHP, DEFI/CHP e ADEMI
Contactos l Contacts [email protected]
Tel: 226 070 300
Fax: 226 070 399
Local das Conferências l Congress Venue
Auditório Prof. Doutor Alexandre Moreira, CHP
(entrada junto à URGÊNCIA - Hospital Santo António)
Summary – Índice
Invited speakers – Comunicações por convite
CC-01 Mechanisms of Primary Ovarian Insuffiency . . . . . . . . . . . . . . . . . . . . . . S6
CC-02 Azoospermia e Oligozoospermia: Características Clínicas e Causas Genéticas . . . . . . . . . . . S7
CC-03 Toxic RNA and Myotonic Dystrophy: From the Dissection of Disease Mechanisms to the Development
of Novel Therapeutic Strategies . . . . . . . . . . . . . . . . . . . . . . . . . . S7
CC-04 Contributo da Neuropatologia para o Diagnóstico das Doenças Neuromusculares . . . . . . . . . . S8
CC-05 Doenças Neuromusculares no Adulto: Abordagem Clínica . . . . . . . . . . . . . . . . . S8
CC-06 New Therapeutic Approaches in Adult Neuromuscular Disorders . . . . . . . . . . . . . . . S9
CC-07 140 Years Since the Identification of Gaucher Disease: What is There Left to Learn? . . . . . . . . . S10
CC-08 Carrier Screening – Will it Be Possible to Eliminate Autosomal Recessive Disorders? . . . . . . . . . S11
Oral communication – Comunicação oral
CO-01 Diagnosis of Neurometabolic Disorders Through Next Generation Sequencing Panels and Bioinformatics Tools:
Two Years of Experience . . . . . . . . . . . . . . . . . . . . . . . . . . . . S12
Poster abstracts – Resumos de posters
P-01 Schwannnomatose Múltipla não Relacionada com NF2 . . . . . . . . . . . . . . . . . . . S13
P-02 Síndrome de Deleção 22q11.2: Diagnóstico em Idade Adulta – Experiência do Centro
de Genética Médica Doutor Jacinto Magalhães - CHP . . . . . . . . . . . . . . . . . . . S14
P-03 Recessive TTN Truncating Mutation Define a Novel Antenatal Severe Form of “Cap-Myopathy” in
Absence of Heart Disease . . . . . . . . . . . . . . . . . . . . . . . . . . . . S15
P-04 Utility of Genetic Panels Based on NGS in the Diagnosis of Childhood Epilepsies . . . . . . . . . . . S15
P-05 Long-Term Follow-Up of Individuals with Williams-Beuren Syndrome: Facilitate Transition from
S16
Pediatric Medicine to Adult Medicine (a Case Report) . . . . . . . . . . . . . . . . . . .
P-06 Aplicação dos Marcadores IRT/PAP/IRT no Rastreio Neonatal da Fibrose Quística . . . . . . . . . . S16
P-07 Translocação 11;22 num Caso de Infertilidade . . . . . . . . . . . . . . . . . . . . . . S17
P-08 Infertility: Importance of Cytogenetic Study . . . . . . . . . . . . . . . . . . . . . . . S17
P-09 Male Infertility and Chromosome Aberrations: Two Case Reports . . . . . . . . . . . . . . . . S18
P-10 Autosome-Autosome Reciprocal Translocation: Implications in the Fertility . . . . . . . . . . . . . S19
P-11 Molecular Profile of Myotonic Dystrophy Type 1 (DM1) in Portuguese Families . . . . . . . . . . .
S19
P-12 High Phenotypic Variability in Two Siblings with Spinal Muscular Atrophy . . . . . . . . . . . . . S20
P-13 Pregnancy in a Patient with Distal Arthrogryposis Type 2B - Clinical . . . . . . . . . . . . . . . S21
P-14 Whole-Exome Sequencing Analysis of Adult Patients with Rare Genetic Diseases: What Have We Learned? . . S22
P-15 Phenotypic Spectrum of DCX Pathogenic Mutations in Females: From Childhood to Adulthood Clinical Onset .
S23
P-16 A Portuguese Family with Cadasil Diagnosis with Anticipation Age of Onset Observed . . . . . . . . . S24
P-17 Lujan-Fryns and Opitz-Kaveggia Syndromes: MED12 Molecular Screening . . . . . . . . . . . . . S25
P-18 Distúrbios Hereditários Raros Revelados pelo Esfregaço de Sangue Periférico . . . . . . . . . . .
S25
P-19 Frontotemporal Dementia and Neuronal Ceroid Lipofuscinosis . . . . . . . . . . . . . . . .
S26
P-20 Forma Adulta da Doença de Pompe em Portugal . . . . . . . . . . . . . . . . . . . . . S27
P-21 Intronic Long Interspersed Nuclear Element (LINE-1) Insertion in the DMD Gene as a Cause of
S28
Becker Muscular Dystrophy . . . . . . . . . . . . . . . . . . . . . . . . . . .
P-22 Doenças Hereditárias do Metabolismo - Afinal Não São Assim Tão Raras no Adulto... . . . . . . . . . S29
Author index – Índice de Autores
. . . . . . . . . . . . . . . . . . . . . . . . . S30
Sponsors and exhibitors – Apoios e expositores
. . . . . . . . . . . . . . . . . . S31
NASCER E CRESCER
revista de pediatria do centro hospitalar do porto
20 de fevereiro de 2015, suplemento I
Invited speakers
Comunicações por convite
CC-01
MECHANISMS OF PRIMARY OVARIAN INSUFFICIENCY
Sophie Christin-Maitre, MD, PhD
Endocrine Unit, AP-HP Hôpital Saint-Antoine, INSERM U933
Université Pierre et Marie Curie
Paris, France
[email protected]
in the oocyte, identified in 6% of POI cases. Other genes such
as elF4ENIF1, STAG 3, HFM1, MCM8 and MCM9 have been
identified recently.
More than 30 candidate genes have been identified so far
in POI. They can be tested by Next Generation Sequencing.
However, the cause of POI is identified in less than 30% of cases.
Studying familial cases of POI should increase our knowledge in
the near future.
Primary ovarian insufficiency (POI) affects 1-2% of the general
population. Patients have primary or secondary amenorrhea,
lasting more than 4 months, occurring before the age of 40. Their
plasma gonadotropins, measured twice, are elevated (FSH > 20
IU/L) with low estradiol levels.
In the absence of previous chemotherapy, radiotherapy and
ovarian surgery, genetic causes of POI need to be investigated.
Many of them are linked to X chromosomal abnormalities,
such as Turner syndrome with 45,X karyotype or 45,X/46,XX
mosaicism. Two copies of two different regions located on Xq,
named POF1 and POF2, are necessary for the maintenance of
ovarian follicles. As those regions escape X inactivation, due
to their haploinsufficiency in Turner syndrome, ovarian follicle
loss is accelerated. Apart from Turner syndrome, the patient’s
karyotype can reveal Xq deletions as well as X autosomal
translocations. In rare cases gene disruption is involved.
However, those Xq regions are poor in gene and a position effect
on autosomal gene might be responsible for follicle depletion.
Taken together, X chromosome abnormalities represent around
10 -15% of POI.
Mental retardation in males from the family, related to fragile
X syndrome, should be searched for, as FMR1 premutations are
identified in 3% and 13% of sporadic cases and familial cases of
POI, respectively. The main mechanism involved is an increased
mRNA inducing ovarian toxicity.
In patients with blepharophimosis, POI can be related to
FOXL2 mutations. POI may also be linked to autoimmunity.
It may rarely belong to Autoimmune Polyendocrinopathy
ECtodermal Dystrophy (APECED) due to AIRE mutations.
More frequently, it is associated with type 1 diabetes, adrenal
insufficiency, hypothyroidism as well as vitiligo, lupus…. As the
genes involved in APS2 and APS4 have not been identified so
far, the autoimmune origin of POI is often difficult to prove. In
less than 2% of cases, POI is due to SF1 mutations.
Many genetic causes of POI are non syndromic. The main
one is NOBOX gene mutations, coding for a protein expressed
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comunicações por convite
XLIV conferências de genética Doutor Jacinto Magalhães
NASCER E CRESCER
CC-02
AZOOSPERMIA E OLIGOZOOSPERMIA:
CARACTERÍSTICAS CLÍNICAS E CAUSAS GENÉTICAS
Nuno Louro, MD
Unidade de Andrologia do Serviço de Urologia, Departamento de
Cirurgia
Centro Hospitalar do Porto, EPE
Porto, Portugal
[email protected]
Em cerca de metade dos casais inférteis existe um factor masculino causal, quer isoladamente quer em conjunto com factores
femininos. Apesar disso, apenas é possível identificar a causa da
infertilidade masculina em 60 a 70% dos casos. A percentagem
de casos potencialmente atribuíveis a causas genéticas é desconhecida, mas é provável que sejam em número significativamente
maior do que aquelas que são possíveis, actualmente, investigar.
A pesquisa de alterações genéticas assume uma dupla importância, por um lado para estabelecer um diagnóstico e, por outro,
pelas questões que pode levantar relativamente à transmissão de
determinados genes e à possibilidade de realizar exame genético
pré-implantação embrionária. As causas genéticas de infertilidade
masculina incluem as anomalias cromossómicas e as mutações
génicas as quais podem influenciar a vários níveis muitos dos processos fisiológicos envolvidos no processo reprodutivo, como a
homeostasia hormonal, a espermatogénese e a qualidade dos
espermatozóides. As anomalias cromossómicas podem ser numéricas ou estruturais. A anomalia cromossómica mais frequente é
o síndrome de Klinefelter, quer na sua forma clássica, quer em
mosaicismos. Outras possibilidades de alterações cromossómicas são a presença de um cariótipo 47,XYY e 46,XX ou as translocações cromossómicas. Também relativamente frequentes são
as alterações estruturais do cromossoma Y (translocações, deleções, inversões e anéis). Destas têm particular importância pela
sua prevalência as microdeleções do braço longo do cromossoma
Y, nomeadamente das regiões AZF (azoospermic factor) a, b e c.
Destas a mais frequente envolve a região AZFc. Ao contrário das
restantes, os portadores desta microdeleção têm habitualmente
espermatozóides no esperma ou no testículo o que coloca o problema da transmissão desta característica para a descendência.
As mutações ligadas ao X ou de genes autossómicos são mais
raras. Destas as mais frequentes são as que originam o síndrome
de Kallman e as mutações do gene do receptor de androgénios
(que levam ao síndrome de insensibilidade aos androgénios, com
manifestações fenotípicas variáveis).
Apesar da importância reconhecida do estudo genético em
casos de infertilidade masculina, vários factores impedem o seu
uso rotineiro, nomeadamente as questões do custo, disponibilidade e dificuldades técnicas. No entanto, dado o potencial de
esclarecer a etiologia do problema, possibilitando a realização de
aconselhamento genético, de forma a que os casais possam tomar
decisões reprodutivas conscientes e informadas, a sua importância
tem sido reconhecida de forma crescente, sendo uma das áreas
da medicina reprodutiva com maior potencial de crescimento no
futuro.
CC-03
TOXIC RNA AND MYOTONIC DYSTROPHY: FROM
THE DISSECTION OF DISEASE MECHANISMS TO THE
DEVELOPMENT OF NOVEL THERAPEUTIC STRATEGIES Mário Gomes-Pereira, PhD
Inserm, Imagine Institute for Genetic Diseases
Paris Descartes University, Sorbonne Paris - Cité University
Paris, France
[email protected]
Myotonic dystrophy (DM) is an autosomal dominant disorder,
caused by the abnormal expansion of non-coding DNA repeats.
The disease is highly multisystemic: in addition to the traditional
muscle and cardiac symptoms, the neurological manifestations
are particularly debilitating, affecting the daily life of patients and
their families.
Transgenic mouse models of the disease have helped
elucidate the molecular mechanisms of disease. Today we
know that repeat-containing expanded RNA accumulates in the
cell nucleus and impairs the activity of RNA-binding proteins,
affecting primarily the regulation of alternative splicing in
multiple tissues. DM is therefore the prototype of toxic RNAmediated spliceopathy. Despite progress in the understanding
of the molecular pathogenesis in skeletal muscle and heart, the
disease pathways in the central nervous system remain unclear.
In the laboratory we have generated a transgenic mouse model
of DM carrying a large repeat expansion. DM mice recreate
molecular features of RNA toxicity in the brain, associated with
relevant behavioural and electrophysiological phenotypes. We
have been using these animals to study the cell populations,
neuronal circuits and molecular pathways primarily affected by
the disease mutation in the central nervous system.
The remarkable progress in the dissection of disease
pathobiology opened new avenues to the rational design of
molecular therapies. Strategies to destroy or neutralise toxic
RNA repeats have successfully rescued the phenotype of DM
mouse models, and are now in clinical trials. The research path
from the identification of disease mutation to the development of
promising therapies took less than 20 years, making of DM not
only a paradigm of RNA toxicity, but also an example of stepby-step dissection of a complex disease mechanism towards
rational therapy design.
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NASCER E CRESCER
CC-04
CONTRIBUTO DA NEUROPATOLOGIA PARA O
DIAGNÓSTICO DAS DOENÇAS NEUROMUSCULARES
Manuel Melo Pires, MD, PhD
CC-05
DOENÇAS NEUROMUSCULARES NO ADULTO:
ABORDAGEM CLÍNICA
Teresa Coelho, MD, PhD
Unidade de Neuropatologia, Departamento de Neurociências
Porto, Portugal
[email protected]
Serviço de Neurofisiologia, Departamento de Neurociências
Porto, Portugal
[email protected]
Desde que iniciamos há cerca de 30 anos o estudo patológico das Doenças Neuromusculares, enormes avanços no
campo da genética foram acontecendo e infelizmente ainda não
existe tratamento específico para grande parte dessas doenças. Cerca de 10000 biópsias de nervo, músculo e pele foram
estudadas na Unidade de Neuropatologia do Hospital de Santo
António e demos formação a todos os principais intervenientes
nesta área: neuropatologistas, neurologistas, neuropediatras,
geneticistas, bioquímicos e outros.
Nesta apresentação fazemos uma revisão das principais
características patológicas do músculo e nervo e como podemos orientar o estudo genético de alguns desses doentes. Queremos salientar também a necessidade do estudo multidisciplinar das doenças neuromusculares criando centros de referência
em hospitais em que as diferentes especialidades envolvidas
estejam bem articuladas.
As doenças neuromusculares são um conjunto muito vasto
de patologias do sistema nervoso periférico, que abrangem o
segundo neurónio motor, as raízes nervosas, os plexos e os
nervos periféricos, a junção neuromuscular e o músculo propriamente dito. Ao contrário do que se passa nas crianças, no
adulto a maioria destas patologias são de natureza adquirida. As
doenças de causa genética são raras mas não deixam de ter um
enorme impacto pessoal e social, não só pela incapacidade que
geram como também pelo seu carácter familiar.
Os sintomas que podem fazer suspeitar de doenças neuromusculares são a fraqueza muscular, apresentada por vezes
como intolerância ao esforço ou cansaço, a perda de sensibilidade, as dores musculares, sobretudo durante ou após
o esforço, as caimbras, a rigidez muscular e as deformações
ósseas. Mas esta lista relativamente pequena de problemas
pode corresponder a situações muito diversas com origem em
qualquer nível do sistema nervoso periférico.
Na verdade o diagnóstico destas doenças tem vindo a tornar-se cada vez mais complexo. Nos últimos trinta anos assistimos ao crescimento explosivo do conhecimento sobre a fisiopatologia destas patologias, incluindo um conhecimento cada
vez mais completo da complexa fisiologia da célula muscular. A
heterogeneidade genética (ou seja a multiplicidade de proteínas
envolvidas e de erros genéticos identificados no mesmo gene)
e a variabilidade fenotípica dentro de cada doença não param
de crescer. Um mesmo gene pode originar expressões fenotípicas diferentes e inclusivamente doença em compartimentos
diferentes. E um mesmo fenótipo pode ser causado pelos mais
variados genes.
Dispomos hoje de instrumentos valiosos para chegar ao
diagnóstico de um determinando tipo de doença neuromuscular. A observação clínica com anamnese, colheita de história
familiar, exame físico e recurso a alguns exames laboratoriais
como doseamento de CK sérico, estudos electromiográficos e
biópsias de nervo e/ou musculo permitem quase sempre chegar a um diagnóstico sindromático. Já o diagnóstico genético
preciso, para além de nem sempre ser possível com o actual
conhecimento, obriga em muitas situações a uma abordagem
multidisciplinar de equipas dedicadas que incluam clínicos, neurofisiologistas, neuropatologistas e geneticistas clínicos e laboratoriais.
Se os diagnósticos genéticos são muitas vezes a última linha
da investigação etiológica não deixa de ser importante ressalvar
que certas patologias clinicamente muito características permitem o seu reconhecimento imediato por clínicos experientes
nesta área. A distrofia miotónica tipo 1 e distrofia facioescapu-
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NASCER E CRESCER
loumeral são dois exemplos de situações em que se pode avançar directamente para o estudo genético, evitando exames mais
invasivos e até indutores de erro de diagnóstico, como é o caso
da biópsia de músculo.
Apresentamos três casos clínicos cujo diagnóstico preciso
só foi possível pelo concurso de uma equipa multidisciplinar
experiente e trabalhando em permanente diálogo.
CC-06
NEW THERAPEUTIC APPROACHES IN ADULT
NEUROMUSCULAR DISORDERS
Teresinha Evangelista, MD, PhD
The Newcastle University, John Walton Muscular Dystrophy Research
Centre
Newcastle upon Tyne, UK
[email protected]
Neuromuscular Disorders (NMD) comprise a diverse group
of inherited and acquired disorders characterized by progressive
muscle weakness and wasting. People of any age can be
affected by NMD and clinical symptoms show a broad spectrum
of severity. NMD often result in severe handicap of the patients.
Pneumonia, cardiac arrhythmias, and cardiac and respiratory
failure are the most frequent causes of death in NMD patients.
Over the past 20 years, research on NMD has undergone
a revolution changing from a largely phenomenological science
into a deeply analytical and technical field. Questions concerning
the primary genes and basic mechanisms involved in NMD
have been answered for a large number of conditions. This
translated in better and earlier diagnosis, better counselling, and
improvement of symptomatic treatments for the majority of NMD
patients with significant gains in life quality and life expectancy.
Unfortunately, cure or a near-normal life on treatment is rather
the exception.
In recent years there has been an increase in novel
therapeutic approaches from testing of existing and new drugs,
to DNA delivery (anti-sense oligonucleotides and plasmid DNA),
gene therapies and stem cells.
The use of viruses for the delivery of genes is now coming
into clinical use. Safer vector designs based on adenovirus or
lentivirus vectors have been developed and in 2012, a market
approval of gene therapy was granted for the viral delivery of
the lipoprotein lipase gene. For DMD, the challenge is the large
size of the mRNA (14 kb) and the need to target all muscles.
Thus dystrophin mini- or micro-genes have been designed and
incorporated with muscle-specific promotors. Gene therapy is
now coming of age for a whole range of different disorders. The
application of exon skipping for the therapy of DMD depends
on the identification of the precise mutation of the patient and
the manipulation of the transcriptome. The correction of the
reading frame in DMD patients should in many cases result in a
BMD phenotype. Splice-modulation therapy aiming at correcting
genetic defects by molecular manipulation of the pre-messenger
RNA is a promising novel therapeutic approach for genetic
diseases.
S9
NASCER E CRESCER
CC-07
140 YEARS SINCE THE IDENTIFICATION OF GAUCHER
DISEASE: WHAT IS THERE LEFT TO LEARN?
Tony Futerman, PhD
Department of Biological Chemistry, Weizmann Institute of Science
Rehovot, Israel
http://www.weizmann.ac.il/Biological_Chemistry/scientist/futerman/
[email protected]
5. Farfel-Becker, T., Vitner, E.B., Kelly, S.L., Bame, J.R., Duan, J.,
Shinder, V., Merrill, A.H. Jr., Dobrenis, K. and Futerman,
A.H. (2013) Neuronal accumulation of glucosylceramide in
a mouse model of neuronopathic Gaucher disease leads to
neurodegeneration. Human Molecular Genetics 23, 843-854.
6. Vitner, E.B., Salomon, R., Farfel-Becker, T., Meshcheriakova,
A., Ali, M., Klein, A.D., Platt, F.M., Cox, T.M. and Futerman,
A.H. (2014) RIP3 as a potential therapeutic target for Gaucher
disease. Nature Medicine 20, 204-209.
Gaucher disease (GD) is an autosomal recessive genetic
disease caused by mutations in the GBA1 gene, which encodes
for the lysosomal hydrolase, glucocerebrosidase (GCase). GD is
normally divided into three phenotypes. Type 1 GD, the most
frequent form of the disease, is considered to have no obvious
neuronopathic features, and the classical hallmark of the disease
is glucosylceramide-laden alternatively-activated macrophages,
known as ‘Gaucher cells’. Type 2 and type 3 GD are
neuronopathic forms (collectively referred to as neuronopathic
GD (nGD)). Glucosylceramide (GlcCer) and glucosylsphingosine
accumulation in the brain leads to neuronal loss in nGD patients
and in nGD mouse models. However, the mode of neuronal
death is not known. We recently demonstrated elevation of proinflammatory cytokines, including interleukin-1 beta (IL-1β) and
tumor necrosis factor-α (TNF α in a mouse model of Gaucher
disease. Our data suggested that neuroinflammation induces
cytotoxic effects in nGD. Inflammation is usually associated with
necrotic rather than apoptotic cell death; cell death via necrosis
leads to microglial activation and pro-inflammatory signaling
cascades. I will now discuss our recent data that shows that
modulating the receptor-interacting protein kinase 3 (Ripk3)
pathway markedly improves neurological and visceral disease
in a mouse model of Gaucher disease. Importantly, Ripk3
deficiency dramatically improved the clinical course of Gaucher
disease mice with increased survival, motor coordination and
salutary effects on cerebral as well as hepatic injury. I will also
discuss additional data demonstrating the involvement of a
number of other pathways in Gaucher disease pathology and
suggest that these too might act as therapeutic targets.
Selected recent references:
1. Vitner, E., Platt, F.M. and Futerman, A.H. (2010) Common
and uncommon pathogenic cascades in lysosomal storage
diseases. J. Biol. Chem. 285, 20423-20427.
2. Farfel-Becker, T., Vitner, E.B., Pressey, S.N.R., Eilam, R.,
Cooper, J.D. and Futerman. A.H. (2011) Spatial and temporal
correlation between neuron loss and neuroinflammation in a
mouse model of neuronopathic Gaucher disease. Hum. Mol.
Genet. 20,1375-1386.
3. Farfel-Becker, T., Vitner, E.B. and Futerman. A.H. (2011)
Animal models for Gaucher disease research. Disease
Models and Mechanisms 4, 746-752.
4. Vitner, E.B., Biton, I., Farfel-Becker, T. and Futerman, A.H.
(2012) Contribution of brain inflammation to neuronal cell
death in neuronopathic forms of Gaucher disease. Brain 135,
1724-1735.
S 10
NASCER E CRESCER
CC-08
CARRIER SCREENING – WILL IT BE POSSIBLE TO
ELIMINATE AUTOSOMAL RECESSIVE DISORDERS?
José Carlos Ferreira, MD, PhD
Medical University of Warsaw
Warsaw, Poland
[email protected]
In a future not far away, anyone can have his/her exome/
genome sequenced for a reasonable price. Lots of debate will
deal with the utility of doing this. But, at least, a likely useful
use of that type of data will be carrier screening. To find out,
prior to conception, if a couple carries recessive disease
causing mutations in the same gene, will make it possible,
using preimplantation or prenatal genetic diagnosis, to identify
embryos or fetuses affected. Current screening methods, for
the vast majority of diseases, rely on family history. The main
drawback of such methods is the need for the birth of an
affected individual – a proband – to identify carriers. Universal
preconceptional screening for all/most recessive disease
causing mutations would reduce dramatically this drawback.
But is it worth the cost? Are there other more cost reasonable
alternatives to exome sequencing based screening? Are there
other more cost reasonable alternatives to universal screening?
That is, population targeted screening?
In this new age of genomics, where exome based screening
is already being offered to anyone who can afford it, it may be
worth to revisit the discussions about carrier screening.
Where there is no screening program, is it reasonable to jump
directly to the exome based screening, wait until it gets more
affordable, or consider the implementation, even temporary, of
other more limited technologies and programs already tested.
Will it be worth? Which perspective should be taken? The public
health perspective or the individual perspective? Are there
differences between the two perspectives?
The history of carrier screening programs and the
discussions that preceded them may be good starting points
in the search for answers to these questions. In the beginning,
carrier screening was based on family history. Later population
targeted preconceptional screening for targeted diseases was
added – sickle cell disease in African ancestry, thalassemias in
Mediterranean ancestry, Tay-Sachs disease in Ashkenazi Jewish
(AJ) ancestry. Then several expansions of number of diseases in
AJ ancestry and universal screening for Cystic Fibrosis occurred.
And more recently, the offer of universal genomic based carrier
screening to anyone who can afford it is the last development
in this process. What were the problems in the implementation
of those programs? What was accomplished? What can be
learned?
This presentation will probably raise more questions than
answer them. Discussion will be expected.
S 11
NASCER E CRESCER
Oral communication
Comunicação oral
CO-01
DIAGNOSIS OF NEUROMETABOLIC DISORDERS
THROUGH NEXT GENERATION SEQUENCING PANELS
AND BIOINFORMATICS TOOLS: TWO YEARS OF
EXPERIENCE
Sofia Gouveia, Ana Fernández-Marmiesse, Iria Otero, Juan
Cocho, Daisy Castiñeiras, José Fraga, Mª Luz Couce
Hospital Clínico Universitario de Santiago de Compostela
Santiago de Compostela, Spain
[email protected]
Genomic research in neurodegenerative and metabolic
disorders is essential for: 1) diagnosis, prognosis and treatment
of these disorders, 2) the application of new preventive
treatments and/or their inclusion in clinical trials of new drugs,
3) provide, within a reasonable period of time, a good family
genetic counselling, and 4) avoid diagnostics odyssey.
The new sequencing technologies require us to leap toward
a new medication and to a new paradigm in which genomic tools
will be essential in the diagnostic process of the patient and, in
many cases for early diagnosis of diseases that could eventually
only be diagnosed by clinical signs and symptoms developed
by the patient.
In recent years, there has been an exponential growth of
knowledge about the genetic basis of disorders. This exponential
growth of knowledge is now one of the biggest challenges faced
by health systems and physicians to their patients. For this
reason, the construction of personal genomic skills is essential,
especially in a reference unit, and is based on already developed
projects (diagnosis of lysosomal disorders, and monogenic
diabetes by massive sequencing).
Since October 2012 we have developed and implemented
in our unit a total of 17 genetic panels based on massive
sequencing technology primarily concerned with the diagnosis
of neuropaediatric and metabolic disorders. So far we have
analyzed a total of 205 patient samples from up to 20 different
hospitals in the country, reaching a diagnosis rate between
40-45%. In this communication we intend not only to relate
~
the results obtained with the application of these paediatric
diagnostic panels, but also the challenges we need to face to
achieve increase speed, efficiency and safety of these diagnostic
tools.
Keywords: metabolic and neuropaediatric disorders,
massive sequencing
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comunicação oral
NASCER E CRESCER
Poster abstracts
Resumos de posters
P-01
SCHWANNNOMATOSE MÚLTIPLA NÃO RELACIONADA
COM NF2
Mónica Bagueixa
Unidade de Saúde de Santa Maria - Unidade Local de Saúde do
Nordeste, Bragança, Portugal
[email protected]
A schwannomatose é uma rara forma genética de neurofibromatose, na qual o doente apresenta dois ou mais
schwannomas não-vestibulares diagnosticados histologicamente.
O diagnóstico implica a exclusão de neurofibromatose
tipo 2 (NF2), através de ressonância magnética nuclear (RMN)
crânio-encefálica de alta resolução de maneira a excluir a
existência de schwannomas vestibulares bilaterais.
O diagnóstico definitivo de schwannomatose aguarda
identificação do locus exato. Para melhor organização clínica e de pesquisa, a National Neuroibromatosis Foundation
propôs um Consenso para uniformizar critérios diagnósticos
dividindo os casos em “confirmados e prováveis e segmentares.
A schwannomatose confirmada pode englobar idade
superior a 30 anos, mais de 2 schwannomas não intradérmicos confirmados histologicamente, sem evidência de tumor
vestibular por RMN de alta resolução e sem mutações NF2.
É apresentado um caso clínico de schwannnomatose
múltipla não relacionada com NF2, incluindo vários tumores
localizados a nível da coluna cervical, dorsal e lombar.
Dada a raridade da situação merece a sua divulgação por
schwannomas múltiplos e intensamente dolorosos.
O objetivo deste estudo é relatar e discutir em detalhes
um caso típico de SCH.
M.A.F.G. de 78 anos, sexo feminino, admtida no Serviço
de Medicina Interna da ULS NE – Unidade Hospitalar de Bragança, a 7 de Junho de 2014 por diminuição da força muscular dos membros inferiores com incapacidade de deambular. História prévia de neoplasia da mama (1995), melanoma
ocular, patologia valvular cardíaca com bioprótese, fibrilação auricular permanente, insuficiência cardíaca congestiva classe 2 NYHA, dislipidemia, patologia osteoarticular
degenerativa e exérese de schwannoma na perna esquerda
(2012). Sem filhos, uma irmã com antecedentes de neoplasia
da mama.
Medicada com rosuvastatina 10 mg id, acenocumarol
conforme esquema, espironolactona 25 mg id, furosemida
40 mg id, lansoprazol 30mg id, bisoprolol 5mg id.
Foram realizados vários exames complementares de
diagnóstico para esclarecimento da incapacidade de deambular.
Atendendo à incapacidade de deambular, provocada
pela dor, decidiu-se com a família pela resseção cirúrgica
da lesão da coluna dorsal (D1-D2) a 1 de Julho de 2014.
Pós-operatório sem intercorrências significativas, apenas
infeção urinária por Klebsiella. O estudo histológico da lesão
revela schwannoma.
Apresenta atualmente melhoria significativa da força
muscular dos membros inferiores e deambula com andarilho.
resumos de posters
S 13
NASCER E CRESCER
P-02
SÍNDROME DE DELEÇÃO 22q11.2: DIAGNÓSTICO
EM IDADE ADULTA – EXPERIÊNCIA DO CENTRO DE
GENÉTICA MÉDICA DOUTOR JACINTO MAGALHÃES –CHP
Natália Tkachenko, Gabriela Soares, Maria João Sá, Ana Rita
Soares, Ana Maria Fortuna
Unidade de Genética Médica, Centro Genética Médica Doutor Jacinto
Magalhães, Centro Hospitalar do Porto - EPE, Porto, Portugal
Conclusão: A identificação de SD 22q11.2, especialmente
em adolescentes e adultos, muitas vezes requer um forte
índice da suspeita. Devido a necessidade de seguimento
multidisciplinar, é essencial reconhecer esta condição com
variabilidade clínica inter e intrafamiliar, o que também possibilita o aconselhamento genético preciso e um diagnóstico
pré-natal específico.
[email protected]
Introdução: A microdeleção 22q11.2 (OMIM #188400/
#192430) é a mais comum, com uma prevalência estimada
de 1 em 4.000 nados-vivos, afetando igualmente o sexo
masculino e feminino. No entanto, a ocorrência real pode ser
maior devido à expressão clínica muito variável. Trata-se da
segunda causa mais comum de atraso do desenvolvimento
e cardiopatia congénita a seguir à Trissomia 21, representa
cerca de 2,4% dos indivíduos com défice cognitivo e 10 a
15% com Tetralogia de Fallot. A deleção 22q11.2 foi descrita
em vários síndromes distintos, tais como Velocardiofacial,
DiGeorge, Takao (ou Conotruncal Anomaly Face Syndrome)
e anomalias conotruncais isoladas; ocasionalmente em diagnóstico clínico de síndromes de Opitz G / BBB autossómico
dominante e cardiofacial de Cayler. Em 1993 foi proposto por
Wilson D. I. et al o termo CATCH 22 com o objetivo de englobar todos estes síndromes ao salientar as principais anomalias associadas à deleção 22q11.2, sendo estas: anomalias
cardíacas do tipo conotruncal, dismorfia facial, hipoplasia/
aplasia do timo, fenda do palato/disfunção velofaríngea e
hipocalcemia. Atualmente, todas patologias acima citadas
ficam coletivamente referidas pela sua etiologia cromossómica: síndrome de deleção 22q11.2 (SD 22q11.2). Devido à
extrema variabilidade clínica, alguns casos serão sub-diagnosticados.
Objetivo: O objetivo deste trabalho foi caracterizar os
doentes da consulta de Genética do CGMJM com diagnóstico do SD 22q11.2 em idade adulta (>18 anos). Foi realizada
a análise retrospetiva dos processos clínicos dos nossos
doentes, motivo de consulta, avaliados antecedentes familiares e pessoais, crescimento, desenvolvimento, morbilidade e
orientação.
Resultado: O diagnóstico de SD 22q11.2 em idade adulta
foi realizado em 9 doentes, 6 do sexo feminino e 3 do sexo
masculino. Os motivos da consulta foram muito variáveis,
nenhum doente foi referenciado por suspeita desta patologia. Oito doentes apresentam dismorfia facial sugestiva de
forma mais ou menos evidente. Todos os doentes têm antecedentes pessoais de dificuldades de aprendizagem e avaliação formal do desenvolvimento cognitivo revelou défice
ligeiro em 5 doentes e moderado em 1. Cardiopatia descrita
em 3 casos, insuficiência velofaríngea em 8 e alterações neuropsiquiátricas em 7 casos. Dois casos são familiares. Todos
os doentes realizaram pesquisa da microdeleção do cromossoma 22, que foi positiva.
S 14
resumos de posters
NASCER E CRESCER
P-03
RECESSIVE TTN TRUNCATING MUTATION DEFINE A
NOVEL ANTENATAL SEVERE FORM OF “CAP-MYOPATHY”
IN ABSENCE OF HEART DISEASE
Ana Fernández-Marmiesse1, M. Carmen Carrascosa-Romero2,
Iria Roca1, Sofia Gouveia1, Mª Luz Couce Pico1
Hospital Clínico Universitario de Santiago de Compostela, Santiago
de Compostela, Spain
2
Neuropediatric Unit, Complejo Universitario Hospitalario de Albacete,
Albacete, Spain
1
[email protected]
Background: Arthrogryposis multiplex congenital
(AMC) is defined as multiple congenital non-progressive
joint contractures involving more than one area of the body.
Amyoplasia, is the most common type of arthrogryposis,
characterized by a generalized replacement of skeletal
muscle by dense fibrous tissue and fat. “CAP myopathy” is
a rare congenital myopathy characterized by cap structures
consisting of disarranged thin filaments with enlarged Z discs,
located at the periphery of the muscle fiber. Four genes have
been associated (ACTA1, TPM2, TPM3 and NEB). To date
TTN gene has never been associated with “CAP-myopathy”.
Methods and results: We report a newborn presenting
with AMC, severe axial hypotonia, and muscular biopsy
compatible with amyoplasia and “CAP-myopathy”. A novel
homozygous truncating mutation (c.38661_38669del) in the
PEVK segment of TTN gene was detected by targeted next
generation sequencing assay.
Conclusion: We report for first time an association
between TTN gene and “CAP-myopathy”, showing that
mutations in this gene should be considered in all congenital
myopathies even if cardiac involvement is absent. We
show also the first described TTN mutation which leads to
totally sarcomere disintegration and placed in an exon only
transcribed in fetal period (isoform IC).
Keywords:
Arthrogryposis
multiplex
congenital,
amyoplasia, “CAP-myopathy”, targeted NGS, TTN-PEVK
P-04
UTILITY OF GENETIC PANELS BASED ON NGS IN THE
DIAGNOSIS OF CHILDHOOD EPILEPSIES
Sofia Gouveia, Ana Fernández-Marmiesse, Iria Otero, Juan
Cocho, Daisy Castiñeiras, José Fraga, Mª Luz Couce
Hospital Clínico Universitario de Santiago de Compostela, Santiago de
Compostela, Spain
[email protected]
Early-onset epileptic encephalopathies constitute a
challenge in daily clinical practice given that, increasingly,
genetic and metabolic causes play an important role,
incorporating new syndromes that require a diagnosis and
a treatment plan. On the other hand, epilepsy prognosis is
conditioned mainly by its aetiology. In recent years, CGHarray studies and epilepsy associated gene studies, have
been added to high-resolution karyotype. Research has led to
the discovery of more than 100 epilepsy genes with important
implications for both research and clinical practice. The most
important clinical application of these findings is the genetic
test, when this information is used to clarify the diagnosis in
patients with epilepsy suspicion (diagnostic test) or is used
to predict the development of disease risk in individuals with
a family history (predictive test). In our paediatric unit it was
designed and implemented an epileptic diagnostic panel
using next generation sequencing technology, which analyze
codifying regions of 110 genes and we have analyzed 52
patients suffering different epilepsy conditions. The average
coverage achieved was ~150-200X, being 0.28% the mean
percentage of bases with coverage <10X. This panel has been
optimized and renewed periodically by adding new genes
related to the epileptic phenotype. We have achieved a high
diagnostic probability (42.3%) in 22 of 52 patients analyzed.
We have considered 8 patients need to be confirmed with
further studies (possible diagnosis) either because of being
a new gene associated with epilepsy or the inability to make
sure about the pathogenicity of the mutation found. We
consider the remaining patients (22) as unsolved (42.3%).
Twelve of the 30 cases with possible or high diagnostic
probability (40%) carried a de novo mutation. Two of the
diagnosed patients had large deletions / insertions involving
one or several genes associated with epilepsy, in which the
second CNV is described as pathogenic and confirmed by
CGH array.
The use of this panel has allowed us a high rate of
diagnosis favouring genetic and family counselling. We also
observed that the use of this technology can lead to the
discovery of new phenotypes in old genes. But the challenge
is to reduce the percentage of non-diagnostic cases through
WES trios studies for the discovery of new genes associated
with epileptic encephalopathy.
Keywords: epilepsy, paediatrics, next generation
sequencing
resumos de posters
S 15
NASCER E CRESCER
P-05
LONG-TERM FOLLOW-UP OF INDIVIDUALS WITH
WILLIAMS-BEUREN SYNDROME: FACILITATE TRANSITION
FROM PEDIATRIC MEDICINE TO ADULT MEDICINE (A
CASE REPORT)
Carla Carmona, Teresa Saraiva, Ana Fortuna
P-06
APLICAÇÃO DOS MARCADORES IRT/PAP/IRT NO
RASTREIO NEONATAL DA FIBROSE QUÍSTICA
Lurdes Lopes, Ana Marcão, Ivone Carvalho, Carmen Sousa,
Helena Fonseca, Hugo Rocha, Laura Vilarinho
Unidade de Rastreio Neonatal, Metabolismo e Genética, Departamento
Genética Humana, Instituto Nacional de Saúde Doutor Ricardo Jorge,
Porto, Portugal
[email protected]
[email protected]
Abstract: Williams – Beuren syndrome (WBS; MIM
194050) is a rare neurodevelopmental disorder caused by a
chromosomal microdeletion at 7q11.23. It results in specific
physical, behavioural, and cognitive abnormalities together
with structural and chemical anomalies in the developing
brain. The physical phenotype includes growth retardation,
a dysmorphic face, heart abnormalities, hyperacusis, infantile
hypercalcaemia, and abnormal gait. The neuropsychological
profile is a striking one, characterized by strengths in certain
complex faculties alongside marked and severe deficits
in cognitive domains. Unlike many others with this kind of
difficulties, WBS tends to have a “gregarious” personality,
being overly friendly with strangers and lack social judgement
skills in general.
Material and Methods: We present the results from
the long-term follow-up of a female adult patient aged 39
years, characterizing her physical and neuropsychological
development. We also characterize her social and educational
context of life.
Results: Our patient revelled during her development the
physical and neuropsychological characteristics of individuals
with WBS. Albeit she had the neurocognitive deficits profile
observed in this syndrome, she always had mental global
levels in the “limit” range, allowing her the access to a good
level of school and professional training.
We also emphasize the importance of all data collected
during her follow-up to facilitate the transition from pediatric
medicine to adult medicine and for her adaptation to different
contexts of life, namely the social and professional contexts.
A Fibrose Quística (FQ) é uma doença genética, com
transmissão autossómica recessiva. Bioquimicamente deve-se à deficiência na proteína Cystic Fibrosis Transmembrane
Condutance Regulator que é codificada pelo gene CFTR,
localizado no cromossoma 7. Estão descritas cerca de 2000
variantes genéticas associadas a esta doença.
Iniciou-se no final de 2013 um estudo piloto integrado
no Programa Nacional de Diagnóstico Precoce (PNDP), que
incluiu 80,000 recém-nascidos (RN).
O aumento da concentração sanguínea da tripsina imunoreactiva (IRT) nos primeiros dias de vida dos RN com FQ
possibilita o rastreio neonatal desta doença. No entanto,
apesar de uma boa sensibilidade (95%), o IRT não é um marcador específico (34-75%) para a FQ, e um rastreio baseado
unicamente neste marcador tem um número elevado de
falsos positivos. Por esta razão, têm sido propostos vários
algoritmos de rastreio, incluindo outros marcadores bioquímicos como a Proteína Associada à Pancreatite (PAP).
Neste estudo, o algoritmo de rastreio utilizado baseiase na determinação do IRT/PAP/IRT em sangue colhido em
papel de filtro, sendo a amostra de sangue a mesma colhida
para as restantes doenças rastreadas. Neste estudo foram
identificadas 680 amostras com valor elevado de IRT ao
rastreio, mas apenas em 272 casos foram solicitadas novas
amostras por apresentarem também aumento de PAP. Esta
estratégia reduziu significativamente os pedidos de segunda
amostra de sangue.
Foram diagnosticados neste estudo 11 doentes, o que
implica uma prevalência ao nascimento de aproximadamente
1:7,200, no entanto este estudo será alargado a mais 80,000
RN para estabelecermos a real prevalência desta patologia
na nossa população.
S 16
resumos de posters
NASCER E CRESCER
P-07
TRANSLOCAÇÃO 11;22 NUM CASO DE INFERTILIDADE
Pedro Botelho, Marta Souto, Regina Arantes, Márcia Martins,
Zélia Gomes, Osvaldo Moutinho, Rosário Pinto Leite
P-08
INFERTILITY: IMPORTANCE OF CYTOGENETIC STUDY
Marta Souto1, Pedro Botelho1, Regina Arantes1, Zélia Gomes2,
Márcia Martins3, Osvaldo Moutinho2,3, Rosário Pinto Leite1
Centro Hospitalar de Trás-os-Montes e Alto Douro, Vila Real, Portugal
1
[email protected]
Introdução: A frequência de anomalias cromossómicas
em homens inférteis varia de 3% a 19%: 3% nos casos de
infertilidade minor e 19% em homens com azoospermia
não-obstrutiva ou com oligozoospermia. As anomalias de
estrutura, nomeadamente, as translocações robertsonianas
e recíprocas podem causar entre 19% a 77% de
espermatozoides com desequilíbrios cromossómicos. Os
autores apresentam o resultado do estudo citogenético
e análise do sémen num casal que recorreu à consulta de
infertilidade.
Material e métodos: O estudo citomorfobioquímico
do esperma foi realizado de acordo com a World Health
Organization Laboratory Manual (5ª edição, 2010). Para
o estudo citogenético ao casal, realizaram-se culturas
sincronizadas, de acordo com os métodos estabelecidos
no laboratório. Foram analisadas 15 metafases e em 15
metafases adicionais excluiu-se a presença de mosaicismos.
A nomenclatura adotada está de acordo com as normas
Internacional System for Human Cytogenetics (ISCN, 2013).
Resultados: Na análise do sémen foi observada uma
baixa concentração espermática (9.8 milhões no total do
ejaculado), com 41% dos espermatozoides imóveis e uma
vitalidade de 64%. No estudo morfológico, apenas 2% os
espermatozoides apresentaram uma morfologia típica. A
análise citogenética do casal revelou um cariótipo masculino
com uma translocação recíproca entre os cromossomas 11 e
22 [46,XY,t(11;22)(q14.2;q13.1)]. Esta anomalia foi observada
em todas as metafases analisadas.
Discussão e Conclusões: A translocação recíproca
entre os cromossomas 11 e 22, com os pontos de quebra
11q23 e 22q11, é a translocação equilibrada mais recorrente.
Os indivíduos portadores são fenotipicamente normais e, na
maioria dos casos, o seu diagnóstico só é detetado devido
a problemas de reprodução. No caso apresentado, os
pontos de quebra envolvidos na translocação são diferentes
dos habitualmente descritos na literatura, sem implicações
fenotípicas aparentes, existindo contudo alterações na
análise do sémen.
Laboratório de Citogenética, Serviço de Genética, Centro Hospitalar
de Trás-os-Montes e Alto Douro, Vila Real, Portugal
2
Serviço de Ginecologia/Obstetrícia, Centro Hospitalar de Trás-osMontes e Alto Douro, Vila Real, Portugal
3
Serviço de Genética, Centro Hospitalar de Trás-os-Montes e Alto
Douro, Vila Real, Portugal
[email protected]
Infertility has been considered by the World Health
Organization as a public health problem. The prevalence of
infertility is increasing in the developed world and affects 1015% of couples in reproductive age.
An etiology for infertility can be found in 80% of cases
with an even distribution of male and female factors, including
couples with multiple factors. Chromosomal abnormalities
affect 5% of infertile males and 4% of the women. The most
frequent anomalies involve the sex chromosomes.
The authors present the cytogenetic results of 221
couples referred to infertility consultation between January
2010 to December 2014. Cytogenetic analysis was performed
according to standard techniques. At least 15 metaphases
were analyzed in each case and additional analysis of 15
metaphases to exclude mosaicism.
Cytogenetic abnormalities were found in 12 cases,
7 in women and 5 in men. Seven balanced reciprocal
translocations were detected. We also observed
aneuploidies involving X chromosome and two markers.
This review corroborates the importance of peripheral blood
karyotype analysis in infertile couples, since it allows a proper
diagnosis and genetic counseling.
Este caso reforça a importância da realização do
cariótipo no estudo da infertilidade de modo a proporcionar
um aconselhamento genético adequado.
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P-09
MALE INFERTILITY AND CHROMOSOME ABERRATIONS:
TWO CASE REPORTS
Fernanda Paula Oliveira1, Natália Oliva Teles1,3, Manuela Mota
Freitas1,3, Gabriela Soares2, Maria da Luz Fonseca e Silva1
Unidade de Citogenética, Centro Genética Médica Doutor Jacinto
2
3
Unidade Multidisciplinar de Investigação Biomédica (UMIB), Instituto
de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto,
Porto, Portugal
1
genetic testing has expanded because of the development
of in vitro fertilization techniques, which makes it obligatory
to consider the possibility of pregnancies in men previously
considered “infertile”. The authors emphasize the importance
of cytogenetic studies as a main approach to evaluate male
infertility and the implications on future generation.
[email protected]
Infertility affects approximately 10%-20% of couples, male
factors contributing about half of the cases. The most frequent
male factors are azoospermia and severe oligozoospermia,
affecting about 10% of individuals; excluding obstructive
causes, genetic etiology is the most common cause of infertility.
The mechanisms that induce male infertility are not clear, but
it is known that a variety of chromosomal alterations may be
responsible. Sex chromosome abnormalities are among the
most frequent problems (e.g., Klinefelter Syndrome; 46,XX
Male Syndrome), but a wide range of structural autosomal
and/or sex chromosome abnormalities are also found (Yq
microdeletions, translocations and inversions). Peripheral
blood karyotype is recommended, but this analysis may need
to be complemented by fluorescence in situ hybridization
(FISH) studies; these include SRY gene probes and/or other
molecular studies, which may be fundamental to establish an
accurate diagnosis.
We report on two patients referred to our Medical
Genetics Centre: Patient 1: male aged 47, with azoospermia;
Patient 2: male aged 36, with severe oligoasthenospermia
and irrelevant family history.
The cytogenetic analysis revealed, in patient 1, 46,XX;
and in patient 2, mos 47,XXY,[2]/46,XY[48]. In patient 1, FISH
for the SRY gene revealed that this gene was located in the
X chromosome.
Males presenting with 46,XX karyotype are referred as
“XX Male Syndrome” or “la Chapelle Syndrome”.
The incidence of this syndrome is very low. One of the
X chromosomes contains the SRY gene and therefore the
patient is phenotypically male, but genetically of female
constitution. Most XX males derive from a crossing over
between Xp and Yp during paternal meiosis, so that the SRY
gene is translocated into the X chromosome.
Klinefelter Syndrome (also known as 47,XXY) is one of
the most common sex chromosome aneuploidy in humans
with a prevalence of about 1 in 600-1000 males. About
20% of patients with Klinefelter Syndrome are mosaics and
47,XXY/46,XY is the most common variant. Genetic testing
in patients with azoospermia or severe oligozoospermia was
not considered very relevant, as the vast majority of these
patients would not be able to reproduce. Currently, however,
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P-10
AUTOSOME-AUTOSOME RECIPROCAL TRANSLOCATION:
IMPLICATIONS IN THE FERTILITY
Sílvia Pires1, Natália Oliva Teles1,2, Nuno Louro3, Maria da Luz
Fonseca e Silva1
P-11
MOLECULAR PROFILE OF MYOTONIC DYSTROPHY TYPE
1 (DM1) IN PORTUGUESE FAMILIES
Márcia E. Oliveira1, Nuno Maia1, Isabel Marques1, Rosário
Santos1,2
Unidade de Citogenética, Centro Genética Médica Doutor Jacinto
2
Porto
3
Serviço de Urologia, Departamento de Cirurgia, Centro Hospitalar do
Porto - EPE, Porto, Portugal
1
1
[email protected]
Unidade de Genética Molecular, Centro Genética Médica Doutor
Jacinto Magalhães, Centro Hospitalar do Porto - EPE, Porto, Portugal;
Porto, Portugal
2
UCIBIO/REQUIMTE, Departamento de Ciências Biológicas,
Laboratório de Bioquímica, Faculdade de Farmácia, Universidade do
Porto, Porto, Portugal
[email protected]
Chromosomal abnormalities have been described as
important causes of male infertility. Structural rearrangements
have been reported as 10 times more frequent among infertile
men than in the general population.
The most common chromosomal abnormality is
Klinefelter’s syndrome, but translocations may also cause
reduction in testicular volume and testosterone level, which
may impact spermatogenesis, resulting in oligozoospermia
or azoospermia and thus male infertility. The involvement of
acrocentric chromosomes and the presence of translocation
breakpoints in close proximity to the centromere are associated
with the most destructive effects on spermatogenesis.
It has been suggested in some publications that the most
likely reason for the development of azoospermia in patients
with reciprocal translocations is a very high proportion
of an association between XY bivalents and quadrivalent
formations in prophase I; the instability in chromosome
segregation during consecutive cell divisions might be a
contributing factor inducing spermatogenic disruption.
The authors present a male patient, aged 41, with
azoospermia and reduced testicular volume, without family
history of infertility.
The karyotype revealed an autosome-autosome
reciprocal
translocation:
46,XY,t(20;22)(q11.21;q11.21).
Chromosomal analysis has become increasingly important
for characterizing possible causes of human infertility, and
has shown that male infertility might be associated with
autosomal chromosome abnormalities. Infertile men who
have chromosomal abnormalities such as translocations but
normal phenotypes have shown that these translocations
may have a devastating impact on spermatogenesis during
meiotic division. In addition, detailed meiotic analysis may
be recommended for each translocation, to obtain better
diagnosis, prognosis and genetic counselling to the patient.
The authors will establish the relationship between male
infertility and chromosomal translocations and compare the
findings of this patient with similar cases described in the
literature.
Myotonic Dystrophy type 1 (DM1), also known as
Steinert disease, is the most common adult form of muscular
dystrophy. It is an autosomal dominant disorder caused by
the expansion of unstable [CTG] repeats in the 3’ untranslated
region (3’UTR) of the gene of myotonic dystrophy protein
kinase (DMPK), located at 19q13.3. The expanded mRNA
products are toxic to cells, affecting normal splicing of other
proteins in different tissues.
Myotonia is the principal manifestation of DM1, but other
organ systems are also affected (e.g. ocular, cardiac and
respiratory). Clinically, DM1 may be phenotipically classified
into four main subtypes: i) mild, ii) classical or adult-onset, iii)
juvenile and iv) congenital. The age of onset and severity are
variable and directly associated with the number of expanded
[CTG] repeats: larger expansions usually result in earlier onset
and a more severe phenotype. Due to the instability of the
expanded [CTG] repeats during transmission, it is common to
observe in affected families a decreasing in age at onset and
an increasing degree of severity in successive generations
(anticipation).
In this work we present the molecular profile of DM1
in affected families studied at our diagnostic service, on a
national basis, since the implementation of the molecular
genetic testing for this disorder in 1997.
All the cases were tested for the presence of expanded
pathogenic alleles. Depending on the size of the expanded
alleles, a combination of three different methods was used
to determine the number of [CTG] repeats in 3’UTR of the
DMPK gene: 1) conventional PCR amplification of the [CTG]repeat region; 2) Triplet repeat-Primed (TP)-PCR and 3)
Southern blotting (SB) technique. Larger expansions (about
[CTG]>100 repeats) are only detected by the last two methods;
moreover, the exact size of these larger expanded repeats
can only be assessed by SB. In order to establish the most
accurate molecular diagnosis, the SB technique is therefore
essential, particularly in the prenatal diagnostic setting.
We also present both clinical and genotype heterogeneity
in the DM1 families, thus demonstrating some of the
characteristics associated with this triplet repeat disorder:
somatic mosaicism, anticipation, influence of gender of the
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transmitting parent and occurrence of inter-generational size
reduction in the expanded repeats.
The aim of the present work is to establish the mutation
profile in patients and their families, where detailed molecular
characterization is fundamental, not only to confirm the
clinical diagnosis and to establish genotype-phenotype
correlations, but also for trial-readiness given the emergent
mutation-based therapies for DM1.
P-12
HIGH PHENOTYPIC VARIABILITY IN TWO SIBLINGS WITH
SPINAL MUSCULAR ATROPHY
Teresa Saraiva1, Jorge Oliveira2,3, Márcia E. Oliveira2,3, Ana
Soares1, Rosário Santos2,3, Ana Fortuna2,3
2
Jacinto Magalhães, Centro Hospitalar do Porto - EPE, Porto, Portugal
3
Porto, Portugal
1
[email protected]
Introduction: Spinal muscular atrophy (SMA) is the
second most common lethal autosomal recessive disease in
caucasians after cystic fibrosis, with an estimated incidence
in Portugal of 1 in 10.800 live births. SMA is a severe
neuromuscular disease characterized by degeneration and
loss of spinal and brain stem motor neurons (lower motor
neurons), resulting in progressive proximal muscle weakness
and atrophy. The disease-causing gene is the survival motor
neuron 1 (SMN1) localized in 5q13. This gene has a highly
homologous copy - SMN2 - differing in only 5 base pairs.
While the SMN2 gene does not compensate entirely the
loss of SMN1 in SMA patients, the number of SMN2 copies
modulates the disease’s severity.
About 95% of patients have a homozygous deletion of
exons 7 and 8 of SMN1. The remaining cases are compound
heterozygotes for the deletion of SMN1 and an intragenic
mutation in the other allele. Clinically SMA is classified
into four subtypes (I – IV) on the basis of age of onset, the
maximum motor function achieved and survivorship. This
classification is useful for prognosis and clinical management.
Intrafamilial phenotypic variability is quite rare, but different
SMA subtypes within the same family have been previously
reported.
Case report: We present a family with two siblings
diagnosed with SMA. They demonstrate a remarkable clinical
variability and were classified with different SMA subtypes.
The first patient, a 25 year-old woman, was referred to
our genetic consultation with proximal limb weakness
and difficulty in walking which started at 22 years of age.
Her brother, 32 years old, is also affected with SMA but
remarkably more severe in weakness. His limb weakness
started at 5 years of age and significantly deteriorated to
lose independent ambulation at the age of 7 years. Molecular
genetic investigations revealed that both sibs have the same
SMN1 genotype: compound heterozygosity for an SMN1
deletion and a novel point mutation [c.460C>T, (p.Gln154*)]
in exon 3 of SMN1. MLPA technique revealed the presence
of two SMN2 copies in both patients.
Conclusion: In this report we demonstrated the presence
of intrafamilial phenotypic variability in two siblings classified
with different SMA subtypes. This variability cannot be
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explained by the number of SMN2 copies, since the siblings
show an identical SMN2 copy number. These results are
suggestive that other modifying factors may influence the
phenotypic variability of SMA, such as gender-related factors,
variants in other loci or modifier genes involved in regulating
alternative splicing. Phenotypic discrepancies in SMA among
siblings are major resources to identify such modifying
factors, which may represent additional therapeutic targets
and contribute towards a better understanding of SMA’s
pathophysiology.
P-13
PREGNANCY IN A PATIENT WITH DISTAL
ARTHROGRYPOSIS TYPE 2B - CLINICAL DIAGNOSIS,
PRENATAL DIAGNOSIS AND GENETIC COUNSELING
Márcia Rodrigues1, Diana Antunes1, Inês Carvalho1, João
Freixo1, Marta Amorim1, Teresa Lourenço1, Ana Bernardo2, Luís
Nunes1
Department of Medical Genetics, Hospital Dona Estefânia, CHLC EPE,
Lisboa, Portugal
2
Prenatal Diagnosis Center, Hospital Dona Estefânia, CHLC EPE,
Lisboa, Portugal
1
[email protected]
Introduction: Arthrogryposis comprises nonprogressive
conditions, which are characterized by multiple joint
contractures. These include a group of autosomal dominant
disorders that mainly involve the distal parts of the limbs
without a primary neurological and/or muscle disease – the
distal arthrogryposis (DA). The various phenotypic forms of
DA are designated DA1 through DA10 and present genetic
heterogeneity. DA type 2B (DA2B) or Sheldon-Hall Syndrome
(SHS, OMIM 601680) is similar to DA1, but affected individuals
tend to have typical craniofacial dysmorphisms. DA2B is
thought to be the most common of the distal arthrogryposis
disorders, with approximately 100 cases described so far.
Aims: We report a clinical case of a young woman with
suspected DA and an ongoing pregnancy with fetal anomalies
also suggestive of arthrogryposis. We hope to provide
evidence that underlines the importance of establishing a
definitive genetic diagnosis in patients with DA, in order to
provide adequate Genetic Counseling (GC) and offer Prenatal
Diagnosis (PND) / Preimplantation Genetic Diagnosis (PGD)
in future pregnancies.
Clinical Case: At 21w+2d of gestational age, the fetal
ultrasound revealed increased nuchal translucency, club foot
and camptodactyly. The 23-year-old patient was then referred
to our outpatient clinic for GC. Amniocentesis was performed
to rule out chromosomal abnormalities: QF-PCR aneuploidy
test and fetal karyotype (46,XX) were both normal. She had
a personal history of multiple distal contractures suggestive
of DA with extensive physiotherapy in childhood and of renal
duplication with recurrent urinary tract infections. No definitive
diagnosis had been established. Because of the uncertainty
of diagnosis and of potential risk of developmental delay, the
couple decided to terminate the pregnancy at 24w. The fetus
was observed by an experienced clinical dysmorphologist. By
combining the clinical features both of the index case and the
fetus, we suspected of DA2B. The molecular testing of TNNI2
gene revealed the heterozygous mutation c.527_529del
(p.K175del), confirming the clinical diagnosis of DA2B. After
terminating this pregnancy, the patient experienced great
psychological distress, with depressive humor and guilt
feelings, and she was referred to Psychiatry. GC was offered
to the couple and they peremptorily refused PND in future
pregnancies, opting for PGD.
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Discussion and Conclusion: This clinical case highlights
the difficulty that clinicians experience in GC when the index
case is a pregnant and the fetus has similar features, but the
clinical diagnosis is not yet confirmed by molecular testing.
One should always keep in mind that the optimal time for
determination of genetic risk and GC regarding prenatal
testing is before pregnancy, even when the prognosis is likely
good as in DA, as well as address associated psychological
aspects.
P-14
WHOLE-EXOME SEQUENCING ANALYSIS OF ADULT
PATIENTS WITH RARE GENETIC DISEASES: WHAT HAVE
WE LEARNED?
Jorge Oliveira1,4, Luís Negrão2, Rute Pereira3,4, Alberto Barros5,
Mário Sousa3,4,5, Rosário Santos1,4,6
2
Consulta de Doenças Neuromusculares, Hospitais da Universidade
de Coimbra, Centro Hospitalar Universitário de Coimbra, Coimbra,
Portugal
3
Departamento de Microscopia, Laboratório de Biologia Celular,
Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto
(ICBAS-UP), Porto, Portugal
4
Porto, Portugal
5
Centro de Genética da Reprodução Prof. Alberto Barros, Porto,
Portugal
1
6
[email protected]
Introduction: Next-generation sequencing (NGS) is
accelerating clinical genetics research and diagnostics, given its
capacity to generate genomic data in a faster and cheaper way.
NGS may avoid the usual stepwise gene-by-gene analysis by
performing targeted resequencing of several loci simultaneously
(gene panels). Wider NGS applications, such as whole-exome
sequencing (WES), may even enable the identification of new
genes associated with human diseases. Nevertheless, WES
applicability is challenging considering the large number of
variants obtained which require specific analytical strategies
and bioinformatic resources. The authors describe the use of
WES in three adult patients, exemplifying its diagnostic potential
but also difficulties encountered during analysis.
Materials and Methods: WES was performed using
the Ion Proton system in five individuals: Case #1- a female
patient with a childhood-onset progressive muscular
dystrophy (35 years of clinical follow-up) and her parents;
Case #2- an infertile male with situs-inversus and total
sperm immotility; Case #3- a male patient presenting limbgirdle muscular dystrophy with onset during early adulthood.
Bioinformatic analysis was performed using several
algorithms for variant annotation, filtering and to identify
autozygosity through runs of homozygosity.
Results and discussion: In case #1, analysis assumed
an autosomal recessive (AR) disease model and focused
on genes implicated in hereditary myopathies. This analysis
suggested the choline kinase beta (CHKB) gene as a
possible candidate, where the detailed scrutiny of sequence
alignments revealed the causal variant (c.1031+3G>C).
Although the mutation was successfully detected its zygosity
was incorrectly called suggesting a possible pitfall in WES.
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A similar approach was used for case #2, resorting to
candidate genes known to be associated with sperm immotility
due to flagellar abnormalities. Variants in additional loci were
also filtered by Gene Ontology. As a result, two novel variants
were identified: a homozygous missense variant (p.Arg35Pro)
in the CCDC103 gene and a novel frameshift variant in the
INSL6 gene (c.262_263delCC).
The experience gathered in the study of these first two
patients was important to delineate the analysis strategy for
case #3, which shall be presented in this work. We propose
a new bioinformatic pipeline for the analysis of AR diseases
using WES, combining variant filtering and autozygosity
mapping.
Concluding remarks: Considering the present state of
the art, WES should be seen as a screening method. There are
technical and analytical limitations to be properly addressed
in WES before incorporating it in routine diagnostics. Our
experience, in line with recent scientific reports, suggests that
WES is presently one of the most efficient and cost-effective
approaches to study highly heterogeneous rare diseases.
P-15
PHENOTYPIC SPECTRUM OF DCX PATHOGENIC
MUTATIONS IN FEMALES: FROM CHILDHOOD TO
ADULTHOOD CLINICAL ONSET
Maria João Sá1,2, Rui Chorão3, Manuela Santos3, Ana Maria
Fortuna1,2, Gabriela Soares1
Unit of Medical Genetics, Centro Genética Médica Doutor Jacinto
2
Multidisciplinary Unit for Biomedical Research, UMIB, Porto, Portugal;
3
Department of Neuropediatrics, Hospital de Santo António/Centro
Hospitalar do Porto - EPE, Porto, Portugal
1
[email protected]
Introduction: DCX-related disorders (MIM#300067)
are caused by pathogenic mutations in the DCX gene
(MIM*300121; Xq23) that result in abnormal neuronal migration
patterns, including isolated lissencephaly sequence (ILS)
and subcortical band heterotopia (SBH; also called double
cortex). Often occurring in males, ILS causes intellectual
disability (ID) and childhood-onset epilepsy. More common in
females, SBH is associated with a broad spectrum of clinical
features, from ID and epilepsy to normal intelligence without
epilepsy. We report two families that illustrate the phenotypic
spectrum of DCX pathogenic mutations in females, from
childhood to adulthood onset.
Patients and methods: Family 1: A 3 years old boy,
born to non-consanguineous parents, presented with
global developmental delay, seizures and microcephaly.
Brain MRI diagnosed fronto-parietal classic lissencephaly.
Sequence analysis of DCX detected a novel likely pathogenic
variant,
c.806G>T,
p.(Gly269Val),
in
hemizygosity.
Segregation analysis confirmed that his mother, who has
mild ID and a frontal simplified gyration pattern shown
by brain MRI, carries this variant in heterozygosity.
Family 2: A 15 years-old girl, born to non-consanguineous
parents, had epilepsy since 4 years old and global
developmental delay. Brain MRI showed SBH and the
previously reported pathogenic variant c.1150C>T, p.(Arg384*)
was identified in DCX gene, in heterozygosity. Her mother,
who carried the same mutation, had epilepsy with onset at 19
years old, an unremarkable brain MRI and normal intelligence.
Discussion: Pathogenic DCX mutations are identified
in approximately 40% of males with classic lissencephaly
(more severe anteriorly than posteriorly), as well as in 85%
of patients with SBH. Given the well-known genotypephenotype correlation in DCX-related disorders, the decision
of testing this gene was made based on the clinical and
cerebral imaging features of the probands.
A novel likely pathogenic variant was identified in DCX,
increasing the genotypic spectrum of mutations in this gene.
DCX mutations were also detected in the probands’ mothers,
who had previously non-investigated mild ID (family 1) and
adult-onset epilepsy (family 2).
DCX-related disorders may not be clinically recognizable
in females due to its clinical heterogeneity. Consequently,
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molecular testing of DCX is warranted in the mothers of
affected children, allowing genetic counselling to at-risk family
members, including prenatal diagnosis and preimplantation
genetic diagnosis. Massive parallel sequencing, either whole
exome or gene panels, of females with ID with or without
epilepsy will likely increase detection of mutations in the DCX
gene.
P-16
A PORTUGUESE FAMILY WITH CADASIL DIAGNOSIS WITH
ANTICIPATION AGE OF ONSET OBSERVED
Maria Lopes-de-Almeida1, Lina Ramos1, Gustavo Cordeiro2,
Rosário Almeida3, Joaquim Sá1, Jorge M Saraiva2,4
Medical Genetics Unit, Hospital Pediátrico, Centro Hospitalar e
Universitário de Coimbra, Coimbra, Portugal;
2
Neurology Unit, Hospital Universitário de Coimbra, Centro Hospitalar e
Universitário de Coimbra, Coimbra, Portugal;
3
Center of Neuroscience and Cell Biology, Faculty of Medicine,
University of Coimbra, Portugal;
4
University Clinic of Pediatrics, Faculty of Medicine, University of
Coimbra, Portugal
1
[email protected]
Introdution: Cerebral Autossomal Dominant Arteriopathy
with Subcortical Infarcts and Leukoencephalopathy
(CADASIL) is a rare hereditary disease characterized by
recurrent transient ischemic attacks, strokes, and vascular
dementia.
Since it is a dominant disease, heterozygous and
homozygous patients are expected to be clinically
indistinguishable. Nevertheless, some homozygous patients
with CADASIL have been reported and in some cases with a
severe phenotype.
Case report: We would like to report a Portuguese family
with inbreeding with diagnosis of CADASIL. It has been found
the p.Arg558Cys mutation in NOTCH3 gene in six members
of this family studied, two of them in homozygosity. One of
the homozygous case present a more severe phenotype
compared with his relatives with an age of onset at 10 years
old. According to this finding, we wonder if the homozygosity
can justify this early age of onset case or its severity.
Discussion: Differences in clinical profile between
homozygous and heterozygous of this family members
and between other CADASIL families with homozygosity
described should be discuss in order to understand if the
homozygosity state increases the pathologic consequences
of the mutation providing a more severe and early phenotype.
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P-17
LUJAN-FRYNS AND OPITZ-KAVEGGIA SYNDROMES:
MED12 MOLECULAR SCREENING
Cathy Paulino1,2*, Isabel Marques1*, Raquel Chaves2, Paula
Jorge1, Rosário Santos1
Jacinto Magalhães, Centro Hospitalar do Porto - EPE, Porto, Portugal;
Porto, Portugal
2
University of Trás-os-Montes and Alto Douro (UTAD), Department of
Genetics and Biotechnology (DGB), Laboratory of Cytogenomics and
Animal Genomics (CAG), Vila Real
* Equal contributors
[email protected]
1
[email protected]
Lujan-Fryns syndrome (LFS - OMIM #309520) is
characterized as a rare form of X-linked Intellectual Disability
(XLID), affecting mostly males. Clinically, the patients can
be recognized by the facial morphology in addition to the
presentation of some of the following features: tall marfanoid
stature, macrocephaly, long hands with hyperextensible
digits, mild general hypotonia and mild to moderate cognitive
deficits with several behavioural problems. Opitz-Kaveggia
syndrome (FGS - OMIM #305450) is also a rare form of XLID.
Patients show a distinctive facial appearance, a tall and
prominent forehead, short stature, small prominente ears with
simplified helical pattern, frontal hair upsweep, hypotonia
and constipation. They frequently present mental retardation
with developmental delay and a distinctive behaviour, with
hyperactive personality and excessive talkativeness. FGS is
characterized by clinical variability and genetic heterogeneity.
There has been a clear association between MED12 gene
and LF and FG syndromes. While in LFS a single recurrent
mutation c.3020A>G in exon 22 was reported, in FGS two
frequent mutations, c.2873G>A and c.2881C>T both located
in exon 21, were previously described. This gene encodes
for the mediator of RNA polymerase II transcription subunit
12, an essential subunit of the mediator complex, interacting
with different developmental pathways and involved in the
regulation of neuronal gene expression.
Until recently, the molecular genetic analysis of MED12 in
our laboratory consisted of sequence analysis of the coding
exons and flanking regions, to screen for variants in seven
of the forty-five exons, considered mutational hot-spots. The
aim of this work is to expand the study of the MED12 gene
to further increase the mutation detection rate, by screening
variants in all exons by PCR-amplification followed by Sanger
sequencing. Preliminary results in our patients revealed the
absence of the previously identified recurrent mutations and
the detection of several variants with unknown significance.
Herein, the pathogenic effect of such variants will be analysed.
P-18
DISTÚRBIOS HEREDITÁRIOS RAROS REVELADOS PELO
ESFREGAÇO DE SANGUE PERIFÉRICO
Cláudia Teixeira1, José Barbot2, Maria Inês Freitas1
Serviço de Hematologia Laboratorial, Departamento de Patologia,
Centro Hospitalar do Porto - EPE, Porto
2
Unidade de Hematologia Pediátrica, Departamento de Medicina,
Centro Hospitalar do Porto - EPE, Porto
1
[email protected]
Introdução: O Esfregaço de Sangue Periférico (ESP)
é uma ferramenta diagnóstica valiosa, que constitui um
complemento importante aos dados clínicos e laboratoriais,
apontando diagnósticos diferenciais e orientando o percurso
diagnóstico. Ocasionalmente, o ESP pode conduzir a
diagnósticos incidentais com grande impacto para o
paciente.
Objetivos: Reportar casos clínicos em que o achado
incidental de alterações morfológicas caraterísticas no ESP
conduziram ao diagnóstico de um distúrbio hereditário
raro. Estas alterações foram encontradas na maioria dos
leucócitos polimorfonucleares do sangue periférico e estão
associadas a distúrbios hereditários específicos.
Resultados: Criança de 4 anos de idade, com história
materna de trombocitopenia crónica inexplicada, com
infeções recorrentes da via aérea superior. Com base na
observação no ESP da tríade de inclusões citoplasmáticas
basofílicas nos neutrófilos, trombocitopenia e plaquetas
gigantes, foi realizado o diagnóstico de anomalia de MayHegglin, um distúrbio autossómico dominante raro que
resulta da mutação do gene MYH9. Subsequentemente
foi realizado ESP à mãe da criança, de 23 anos de idade,
e foram observadas inclusões leucocitárias similares, tendo
sido identificada a anomalia de May-Hegglin como a causa
da trombocitopenia.
Grânulos citoplasmáticos gigantes foram encontrados
incidentalmente, no exame do ESP, nos neutrófilos,
eosinófilos e outros granulócitos de um paciente de 3 anos de
idade durante um episódio de infeção pelo vírus Epstein-Barr
confirmada serologicamente. Estas alterações leucocitárias
são patognomónicas da Síndrome de Chediak-Higashi
(SCH), um distúrbio autossómico recessivo raro associado a
infeções piogénicas recorrentes, alterações da coagulação e
anormalidades neurológicas progressivas. Apesar do curso
benigno da infeção, este diagnóstico foi confirmado pela
presença de um padrão anormal de pigmentação do cabelo
à microscopia ótica. O estudo genético identificou uma
mutação missense no gene CHS1/LYST, compatível com
uma forma mais leve do SCH.
Criança de 10 anos de idade, referida à consulta de
Pediatria por atraso de crescimento e queixas articulares.
O achado de grânulos intensamente azurofílicos (grânulos
de Alder-Reilly) nos leucócitos do sangue periférico sugeriu
o diagnóstico de Mucopolissacaridose (MPS). A análise
do glicosaminoglicano urinário e da atividade enzimática
resumos de posters
S 25
NASCER E CRESCER
confirmou o diagnóstico de MPS VI, um distúrbio do
armazenamento lisossomal raro com um padrão de herança
autossómica recessiva.
Conclusão: Os nossos resultados sublinham a
importância do reconhecimento do ESP como uma
ferramenta diagnóstica valiosa, que fornece informação
relevante para o diagnóstico de diversos distúrbios
hereditários hematológicos e não-hematológicos.
P-19
FRONTOTEMPORAL DEMENTIA AND NEURONAL CEROID
LIPOFUSCINOSIS
Ana Luísa Carvalho1, Lina Ramos1, Maria Margarida Venâncio2,
Isabel Santana3, Carmo Macário4, Rosário Almeida5, Jorge
Saraiva6
Universitário de Coimbra, Coimbra, Portugal
2
Universitário de Coimbra, Coimbra, Portugal and Department of
Medical Genetics, Faculty of Medicine, University of Coimbra,
Portugal
3
Department of Neurology, Hospitais da Universidade de Coimbra,
Centro Hospitalar e Universitário de Coimbra, Coimbra, Portugal and
Faculty of Medicine, University of Coimbra, Portugal
4
Department of Neurology, Hospitais da Universidade de Coimbra,
Centro Hospitalar e Universitário de Coimbra, Coimbra, Portugal
5
CNC, Center for Neuroscience and Cell Biology, University of
Coimbra, Portugal
6
Universitário de Coimbra, Coimbra, Portugal and University Clinic of
Pediatrics, Faculty of Medicine, University of Coimbra, Portugal
1
[email protected]
Introduction: Frontotemporal dementia is the second
most frequent form of early-onset dementia. Its molecular
basis is heterogeneous. Heterozygous mutation in the
progranulin gene (GRN) is a frequent cause of this disease,
with autossomal dominant inheritance. Recently, Smith et
al. presented two brothers with adult-onset neuronal ceroid
lipofuscinosis and homozygous mutation in the GRN gene
(c.813_816del (p.Thr272Serfs*10)). This type of neuronal
ceroid lipofuscinosis was designated type 11 (CLN11).
Case report: We report one family with frontotemporal
dementia with molecular diagnosis: heterozygous for
g.22632264dupGT (p.Ser301Cysfs*60) mutation in the
GRN gene. One member of this family presented with
progressive visual failure at 25 years, followed by dystonia
with muscle weakness, multifocal myoclonus and dysarthria.
Plasma progranulin values are undetectable. The molecular
analysis of GRN gene revealed a homozygous mutation
g.22632264dupGT (p.Ser301Cysfs*60) confirming the
diagnosis of CLN11.
Comment: Mutations in specific genes usually
determine important phenotypes in either the heterozygous
or homozygous state. In this two families, with mutations
in the GRN gene, two clinical distinct neurological
disorders are present: frontotemporal dementia at
heterozygous and CLN11 at homozygous state.
A deletion was present in Smith et al. previously reported
family. The family reported by the authors is the first with
homozygous state for a duplication in the GRN gene.
S 26
resumos de posters
NASCER E CRESCER
P-20
FORMA ADULTA DA DOENÇA DE POMPE EM PORTUGAL
Carla Caseiro1, Francisco Laranjeira1, Elisabete Silva2, Helena
Ribeiro2, Célia Ferreira2, Fernanda Pinto2, Isaura Ribeiro1,2,
Domingos Sousa1, Sónia Rocha1, Eugénia Pinto1, Sara
Pacheco1, Dulce Quelhas1,2, Lúcia Lacerda1,2
nóstico. A consciencialização dos clínicos para esta possibilidade, de fácil e barato diagnóstico laboratorial, poderá levar
a uma diminuição do intervalo de tempo para obtenção do
diagnóstico, com melhorias no prognóstico dos doentes sintomáticos e de outros familiares ainda assintomáticos.
Unidade de Bioquímica Genética, Centro Genética Médica Doutor
2
Porto, Portugal
1
[email protected]
Introdução: A doença de Pompe (OMIM#232300) é uma
doença lisossómica, de transmissão autossómica recessiva,
causada por deficiência da enzima alfa-glucosidase ácida
(GAA), codificada pelo gene GAA localizado no cromossoma
17q25. A função alterada desta enzima provoca acumulação
intralisossomal de glicogénio em vários tecidos, especialmente no músculo esquelético. A doença de Pompe tem um
espectro clínico muito amplo, podendo apresentar-se com
início precoce, durante o primeiro ano de idade, rapidamente
progressivo, até início tardio - formas juvenis ou adultas,
consideradas raras - com uma evolução lentamente progressiva. Nas formas adultas o quadro clínico clássico é de uma
miopatia progressiva sendo o envolvimento cardíaco raro.
As biópsias musculares normalmente revelam aumento de
glicogénio mas uma biópsia normal não exclui a patologia.
Existe terapêutica enzimática de substituição (ERT) desde
2006, que nas formas de início tardio asseguram a funcionalidade muscular e a qualidade de vida. O início precoce
do tratamento, antes da destruição da arquitetura muscular,
permite melhores resultados.
Objectivo: Realizar uma análise retrospetiva dos doentes
de Pompe, forma adulta, diagnosticados no Centro de Genética Médica Doutor Jacinto Magalhães – Centro Nacional de
Referência - desde 1984.
Métodos: O diagnóstico laboratorial da Doença de
Pompe poderá ser dividido em 3 fases: screening inicial pela
determinação da atividade da GAA em DBS (Dried Blood
Spots), confirmação do diagnóstico pela determinação da
atividade da GAA em leucócitos totais ou fibroblastos cultivados e, finalmente, diagnóstico molecular para identificação
das mutações causais no gene GAA.
Resultados: Dos 47 doentes de Pompe diagnosticados
até ao momento, 23 casos, pertencentes a 17 famílias, apresentam a forma adulta da doença.
São apresentados os dados clínicos, bioquímicos e de
genética molecular destes doentes, incluindo os que estão
sujeitos a ERT, estabelecendo-se também as correlações
genótipo-fenótipo.
Conclusões: As formas adultas da Doença de Pompe
são difíceis de detetar clinicamente porque se confundem
com outras doenças neuromusculares e, pela sua prevalência, serem raramente equacionadas no algoritmo de diag-
resumos de posters
S 27
NASCER E CRESCER
P-21
INTRONIC LONG INTERSPERSED NUCLEAR ELEMENT
(LINE-1) INSERTION IN THE DMD GENE AS A CAUSE OF
BECKER MUSCULAR DYSTROPHY
Ana Gonçalves1,5, Teresa Coelho2, Jorge Oliveira1,5, Emília
Vieira1,5, Ricardo Taipa3, Manuel Melo Pires3, Elsa Bronze da
Rocha4, Rosário Santos1,4,5
Jacinto Magalhães - Centro Hospitalar do Porto, EPE - Porto, Portugal
2
Consulta de Neurologia/Doenças Neuromusculares, Centro Hospitalar
do Porto - EPE, Porto, Portugal
3
Unidade de Neuropatologia, Centro Hospitalar do Porto - EPE, Porto,
Portugal
4
5
Porto, Portugal
1
out-of-frame transcript, a residually expressed wild-type
transcript was also detected, thereby explaining the milder
phenotype in this patient.
To our knowledge this is the first report ever of
dystrophinopathy caused by an intronically placed L1
element. Besides representing an exceptional contribution
towards widening the DMD gene mutation spectrum, this
study highlights the importance of conducting mRNA studies
in as yet uncharacterized BMD/DMD patients. This holds true
even considering some of the most recent state-of-the-art
screening approaches, based on next-generation sequencing
technology, where this type of mutation may ultimately fail to
be detected.
[email protected]
Long interspersed nuclear elements (LINE-1 or L1) are
the most abundant retrotransposable elements accounting
for nearly 17% of the human genome. These elements
can be randomly incorporated in the genome, therefore
having an important role in its plasticity and in generating
structural genetic variants. It has been demonstrated that
L1 retrotransposon activity may occasionally cause genetic
diseases. To date, only four disease-causing L1 elements
have been described in the dystrophin (DMD) gene; three
inserted in exons 44, 48 and 67, in patients with a Duchenne
muscular dystrophy (DMD) phenotype, and one detected
in the 5´untranslated region, in two apparently unrelated
Japanese families with X-linked dilated cardiomyopathy.
We report a 48 year old man with a clinical diagnosis
of Becker muscular dystrophy (BMD), in 2001, without
molecular confirmation by multiplex PCR and Southern-Blot
analysis, and whose diagnosis was recently revisited because
his daughter is considering pregnancy. A second molecular
study, resorting to multiplex ligation-probe amplification
(MLPA) analysis and genomic DMD gene sequencing, again
failed to detect abnormalities. A new muscle biopsy showed
dystrophic features with irregular labeling for dystrophin on
immunohistochemical analysis, suggesting dystrophinopathy.
With the intention of unveiling a genetic defect that
might be refractory to the previous diagnostic techniques,
muscle-derived DMD transcripts were sequenced in their
entirety. Results revealed an insertion of 103 nucleotides
between exons 51 and 52, which showed no homology to the
gene’s reference sequence. Extensive bioinformatic analysis
(homology search and splice-site/branch-point analysis) and
sequential direct sequencing enabled the discovery of a deepintronic insertion of an L1 element, in intron 51. This extremely
rare mutational event resulted in the partial exonization of the
L1 plus 5 nucleotides of intron 51. In addition to the aberrant
S 28
resumos de posters
NASCER E CRESCER
P-22
DOENÇAS HEREDITÁRIAS DO METABOLISMO – AFINAL
NÃO SÃO ASSIM TÃO RARAS NO ADULTO...
Sónia Rocha1, Francisco Laranjeira1, Carla Caseiro1, Isaura
Ribeiro1,2, Eugénia Pinto1, Célia Ferreira1, Helena Ribeiro1, Dulce
Quelhas1,2, Lúcia Lacerda1,2
o acesso a estas análises altamente específicas a médicos
de diversas especialidades e de todo o país. Contudo, ainda
não é possível afirmar se esse número se deve a um diagnóstico tardio ou a um efetivo aparecimento de sintomatologia
na idade adulta.
Unidade de Bioquímica Genética, Centro Genética Médica Doutor
2
Porto, Portugal
1
[email protected]
Introdução: A Unidade de Bioquímica Genética do Centro de Genética Médica Doutor Jacinto Magalhães (UBG-CGMJM) desenvolve atividade integrada de Assistência,
Investigação e Formação no diagnóstico de cerca de 100
Doenças Hereditárias do Metabolismo (DHM), pertencentes
aos grupos de doenças Lisossomais, Peroxissomais e Défices Congénitos da Glicosilação.
Estas patologias apresentam fenótipos clínicos muito
diversos, que vão desde formas neonatais, na sua maioria
muito graves, até formas com apresentação mais tardia, que
pode ser infantil, juvenil ou adulta, esta última clinicamente
mais moderada.
Objetivos: Neste trabalho apresenta-se a casuística
obtida em 30 anos de estudo de cerca de 16 000 casos com
suspeita clínica de DHM, pretendendo chamar a atenção
para a apresentação no doente adulto.
Métodos: O diagnóstico pré e pós-natal da maioria das
doenças estudadas é efetuado a 3 níveis de estudo, ou seja,
pelo rastreio dos metabolitos acumulados, pela determinação da deficiência metabólica e finalmente pela identificação
das mutações causais.
Resultados: Foram diagnosticados cerca de 1000
doentes no total das patologias estudadas, dos quais 30%
em idade adulta, distribuídos por 30 patologias. Embora a
doença de Gaucher continue a ser a patologia lisossomal
mais frequente (de 135 diagnósticos efetuados, 94 são formas adultas), nos últimos anos aumentou muito o número de
diagnósticos de doença de Fabry, tornando-a na segunda
patologia em número de doentes (133 diagnósticos efetuados) e mesmo na primeira em número de formas adultas (119
diagnósticos). São apresentados os dados referentes à distribuição por patologias e dados epidemiológicos.
Conclusão: Embora individualmente raras, coletivamente constituem um grupo significativo de doentes.
Com o desenvolvimento de abordagens terapêuticas ganha
maior premência o diagnóstico precoce como forma de minimização do desenvolvimento de lesões irreversíveis.
O número significativo de adultos diagnosticados a partir
de 2009, deve-se à crescente sensibilização de especialidades médicas para além da pediatria.
Os projetos de rastreio em sangue seco capilar têm
contribuído sobremaneira para esta evolução, permitindo
resumos de posters
S 29
NASCER E CRESCER
Author index
Índice de autores
Almeida R., P-16, P-19.
Amorim M., P-13.
Antunes D., P-13.
Arantes R., P-07, P-08.
Bagueixa M., P-01.
Barbot J., P-18.
Barros A., P-14.
Bernardo A., P-13.
Botelho P., P-07, P-08.
Bronze da Rocha E., P-21.
Carmona C., P-05.
Carrascosa-Romero M. C., P-03.
Carvalho A. L., P-19.
Carvalho Ivone, P-06.
Carvalho Inês, P-13.
Caseiro C., P-20, P-22.
Castiñeiras D., CO-01, P-04.
Chaves R., P-17.
Christin-Maitre S., CC-01.
Cocho J., CO-01, P-04.
Coelho T., CC-05, P-21.
Cordeiro G., P-16.
Couce ML., CO-01, P-03, P-04.
Evangelista T., CC-06.
Fernández-Marmiesse A., CO-01, P-03,
P-04.
Ferreira C., P-20, P-22.
Ferreira JC., CC-08.
Fonseca e Silva M. L., P-09, P-10.
Fonseca H., P-06.
Fortuna A. M, P-02, P-05, P-12, P-15.
Fraga J., CO-01, P-04.
Freitas M.I., P-18.
S30
Freixo J., P-13.
Futerman T., CC-07.
Gomes Z., P-08.
Gomes-Pereira M., CC-03.
Gonçalves A., P-21.
Gouveia S., CO-01, P-03, P-04.
Jorge P., P-17.
Lacerda L., P-20, P-22.
Laranjeira F., P-20, P-22.
Lopes L., P-06.
Lopes de Almeida M., P-16.
Lourenço T., P-13.
Louro N., CC-02, P-10.
Macário C., P-19.
Maia N., P-11.
Marcão A., P-06.
Marques I., P-11, P-17.
Martins M., P-07, P-08.
Melo Pires M., CC-04, P-21.
Mota Freitas M., P-09.
Moutinho O., P-07, P-08.
Negrão L., P-14.
Nunes L., P-13.
Oliva Teles N., P-09, P-10.
Oliveira F. P., P-09.
Oliveira J., P-12, P-14, P-21.
Oliveira M. E., P-11, P-12.
Otero I., CO-01, P-04.
Pacheco S., P-20.
Paulino C., P-17.
Pereira R., P-14.
Pinto E., P-20, P-22.
Pinto F., P-20.
índice de autores
Pinto Leite R., P-07, P-08.
Pires S., P-10.
Quelhas D., P-20, P-22.
Ramos L., P-16, P-19.
Ribeiro H., P-20, P-22.
Ribeiro I., P-20, P-22.
Roca I., P-03.
Rocha H., P-06.
Rocha S., P-20, P-22.
Rodrigues M., P-13.
Silva E., P-20.
Soares A., P-12.
Soares A. R., P-02.
Soares G., P-02, P-09, P-15.
Sousa C., P-06.
Sousa D., P-20.
Sousa M., P-14.
Souto M., P-07, P-08.
Taipa R., P-21.
Teixeira C., P-18.
Tkachenko N., P-02.
Venâncio M. M., P-19.
Vieira E., P-21.
Vilarinho L., P-06.
Sá J., P-16.
Sá M. J., P-02, P-15.
Santana I., P-19.
Santos M., P-15.
Santos R., P-11, P-12, P-14, P-17, P-21.
Saraiva J., P-16, P-19.
Saraiva T., P-05, P-12.
NASCER E CRESCER
Sponsors and exhibitors
Apoios e expositores
apoios e expositores
S31
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Suplemento - Revista Nascer e Crescer

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