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Técnicas De Análise Genômica Permitem Estabelecer O Diagnóstico Etiológico De Crianças Com Baixa Estatura De Causa Desconhecida

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Técnicas De Análise Genômica Permitem Estabelecer O Diagnóstico Etiológico De Crianças Com Baixa Estatura De Causa Desconhecida Thais Kataoka Homma Técnicas de análise genômica permitem estabelecer o diagnóstico etiológico de crianças com baixa estatura de causa desconhecida Tese apresentada à Faculdade de Medicina da Universidade de São Paulo para obtenção de título de Doutor em Ciências Programa de Endocrinologia Orientador: Prof. Dr. Alexander Augusto de Lima Jorge Coorientadora: Prof. Dra. Alexsandra Christianne Malaquias de Moura Ribeiro São Paulo 2019 Dados Internacionais de Catalogação na Publicação (CIP) Preparada pela Biblioteca da Faculdade de Medicina da Universidade de São Paulo ©reprodução autorizada pelo autor Homma, Thais Kataoka Técnicas de análise genômica permitem estabelecer o diagnóstico etiológico de crianças com baixa estatura de causa desconhecida / Thais Kataoka Homma. -- São Paulo, 2019. Tese(doutorado)--Faculdade de Medicina da Universidade de São Paulo. Programa de Endocrinologia. Orientador: Alexander Augusto de Lima Jorge. Coorientadora: Alexsandra Christianne Malaquias de Moura Ribeiro. Descritores: 1.Transtornos do crescimento 2.Insuficiência de crescimento 3.Retardo do crescimento fetal 4.Genética 5.Sequenciamento completo do exoma 6.Hibridização genômica comparativa USP/FM/DBD-105/19 Responsável: Erinalva da Conceição Batista, CRB-8 6755 Esse estudo foi realizado na Unidade de Endocrinologia Genética (LIM/25), Laboratório de Hormônios e Genética Molecular (LIM/42) e Laboratório de Sequenciamento em Larga Escala (SELA). Disciplina de Endocrinologia, Faculdade de Medicina da Universidade de São Paulo. Contou com o apoio financeiro da Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) através dos processos 2013/03236–5 e 2015/26980-7 e com apoio do Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) através dos processos 304678/2012–0. Aos meus avós Takeshiro e Yoshime Homma e Toshio e Juvenil Kataoka in memoriam Exemplos de força, coragem, integridade e determinação AGRADECIMENTOS Gostaria de agradecer a todos que direta ou indiretamente contribuíram para a realização desse trabalho. Ao meu orientador Prof. Alexander Jorge e à minha coorientadora Profa. Alexandra Malaquias agradeço imensamente a confiança e atenção dedicada. Muito obrigada pelo acolhimento, carinho e amizade. Obrigada por todos os ensinamentos e incentivos. Meu agradecimento especial também ao Prof. Ivo Arnhold que sempre se fez presente em toda a trajetória desse estudo, colaborando com preciosas sugestões. Obrigada por toda a sua generosidade, auxílio e orientação em todos os trabalhos e apresentações. Ao grupo de Genética do Instituto da Criança, meu profundo agradecimento à Dra Rachel Honjo, Dra. Sofia Sugayama, Dra. Chong Ae Kim, Dra Débora Bertola e a todos os residentes que sempre se mostraram disponíveis e participantes no trabalho, nos ajudando com encaminhamentos e avaliação dos pacientes. Muito obrigada! Aos colaboradores Dra. Ana Krepischi, Dra. Tatiane Furuya; Dra Silva Costa, Dra. Rosimeire Roela, Dra Ana Canton, Dr. Antônio Lerário, Dr. Andrew Dauber, Dr. Paulo F. Collet-Solberg, Dra. Aline Leal e Dra. Elvira Velloso pelo apoio e participação nesse estudo. Gostaria de agradecer ainda a todos os colegas da pós-graduação e funcionários do LIM/42, LIM/25 e SELA. Muito obrigada pela amizade, companheirismo e atenção disponibilizada. Um agradecimento especial à Bruna Lucheze, Mariana Funari, Amanda Narcizo e Dra Mirian Nishi, que contribuíram imensamente para a realização deste estudo com os trabalhos no laboratório; e às colegas Edoarda Vasco, Renata Noronha, Gabriela Vasques, Renata Scalco, Thaís Lima, Nathália Albuquerque, Marilena Nakaguma, Fernanda Corrêa, Isabela Biscotto e Nathália Lisboa, pela companhia nos ambulatórios, congressos e nos intervalos de almoço. Obrigada pelo apoio e cumplicidade. A todos os Professores do curso de Pós-Graduação em Endocrinologia da Faculdade de Medicina da Universidade de São Paulo (FMUSP) e aos meus Professores da residência de Endocrinologia Pediátrica da Santa Casa de São Paulo (FSCMSP) que com dedicação, entusiasmo e o dom de ensinar, são nosso grande exemplo para que sempre busquemos novos conhecimentos. Muito obrigada pelo acolhimento, ensinamentos. e pela oportunidade de convívio, que certamente contribuíram para o meu enriquecimento profissional e pessoal. À Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) e ao Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), meus sinceros agradecimentos pelo apoio oferecido para que o projeto fosse realizado. E, sobretudo, gostaria de agradecer minha família, em especial aos meus pais Alfredo e Liete Homma, “minhas irmãs” Érika, Amanda, Ivete e Julyana, e meu noivo Rafael, pelo amor, apoio e incentivos constantes que me permitiram realizar mais esse sonho. E aos meus amigos que sempre estiveram ao meu lado durante todo esse processo, especialmente Maissara, Juliana, Lara, Débora, Bruna, Aleks, Paulinha, Fabíola e Antonette; pessoas maravilhosas que tenho o imenso prazer de conviver. Finalmente, meu profundo e infinito agradecimento aos pacientes que são a nossa maior fonte de inspiração e ensinamentos. Muito obrigada pela colaboração essencial nesse estudo. NORMAS ADOTADAS Esta tese está de acordo com as seguintes normas, em vigor no momento desta publicação: Referências: adaptado de International Committee of Medical Journals Editors (Vancouver). Universidade de São Paulo. Faculdade de Medicina. Divisão de Biblioteca e Documentação. Guia de apresentação de dissertações, teses e monografias. Elaborado por Anneliese Carneiro da Cunha, Maria Julia de A. L. Freddi, Maria F. Crestana, Marinalva de Souza Aragão, Suely Campos Cardoso, Valéria Vilhena. 3a ed. São Paulo: Divisão de Biblioteca e Documentação; 2011. Abreviaturas dos títulos dos periódicos de acordo com List of Journals Indexed in Index Medicus. SUMÁRIO RESUMO ABSTRACT INTRODUÇÃO ...................................................................................................... 1 OBJETIVOS .......................................................................................................... 3 RESULTADOS ...................................................................................................... 4 CAPÍTULO 1 ......................................................................................................... 5 - Recurrent copy number variants associated with syndromic short stature of unknown cause CAPÍTULO 2 ……………………………………………………………………. 18 - Rare genetic disorders in prenatal onset syndromic short stature identified by exome sequencing. CAPÍTULO 3 ……………………………………………………………………. 42 - Homozygous loss of function BRCA1 variant causing a Fanconi-anemia- like phenotype, a clinical report and review of previous patients CAPÍTULO 4 ......................................................................................................... 48 - Multigene sequencing analysis of children born small for gestational age with isolated short stature DISCUSSÃO .......................................................................................................... 65 CONCLUSÕES ...................................................................................................... 71 ANEXO ................................................................................................................... 72 - Aprovação pelo Comitê de Ética em Pesquisa REFERÊNCIAS ..................................................................................................... 76 RESUMO Homma TK. Técnicas de análise genômica permitem estabelecer o diagnóstico etiológico de crianças com baixa estatura de causa desconhecida. [tese]. São Paulo: Faculdade de Medicina, Universidade de São Paulo; 2019. INTRODUÇÃO: Crianças com baixa estatura constituem um grupo heterogêneo. Em uma parcela dos casos, o mecanismo envolvido nesse processo decorre de alterações genéticas. OBJETIVO: Realizar uma investigação clínica e genético-molecular de um grupo de pacientes com baixa estatura de causa desconhecida. MÉTODOS: Selecionamos crianças com baixa estatura persistente (escore-Z de altura ≤ -2 para idade e sexo) de causa desconhecida para avaliação genômica. O estudo foi dividido em 2 etapas: 1ª etapa – avaliação de 229 pacientes com baixa estatura sindrômica [baixa estatura associada a outros achados dismórficos (atraso de desenvolvimento neuropsicomotor e/ou déficit intelectual, presença de dismorfismos faciais e/ou outras malformações)] por cariótipo molecular (aCGH/SNPa); 2ª etapa: avaliação de 99 crianças com baixa estatura persistente, nascidas pequenas para idade gestacional (PIG - escore-Z de peso e/ou comprimento ao nascer ≤ -2 para idade gestacional) e classificação de acordo com a presença ou ausência de dismorfismos associados. Essas crianças foram divididas em dois grupos: baixa estatura sindrômica (n=44) e baixa estatura isolada (n=55). Pacientes com baixa estatura sindrômica foram avaliados por sequenciamento exômico (WES). Pacientes com baixa estatura isolada foram avaliados através de painel gênico (n = 39) ou WES (n = 16). RESULTADOS: 1ª etapa: 32 (14%) pacientes com baixa estatura sindrômica apresentaram variações no número de cópias (CNVs) patogênicas ou possivelmente patogênicas. Sete delas são recorrentes em outros estudos e são responsáveis por cerca de 40% de todas as CNVs patogênicas/possivelmente patogênicas encontradas em pacientes com baixa estatura de causa desconhecida. 2ª fase: Dentre os 99 pacientes avaliados com baixa estatura nascidos PIG, foram encontradas 23 variantes patogênicas/possivelmente patogênicas em genes já associados à distúrbios
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