Orientador: Antonio Fernando Ribeiro / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-19T10:15:56Z (GMT). No. of bitstreams: 1 Marsom_FernandoAugustodeLima_M.pdf: 12182231 bytes, checksum: e98655cb1347b5ba1ffbceac13daebb0 (MD5) Previous issue date: 2011 / Resumo: Introdução: A fibrose cística (FC) se manifesta clinicamente com várias formas de gravidade, moduladas por diferentes genótipos e o meio ambiente. Enquanto as classes das mutações no gene CFTR ("Cystic Fibrosis Transmembrane Conductance Regulator") estão bem definidas, como moduladores de gravidade da FC, os polimorfismos continuam controversos. Objetivo: Verificar se polimorfismos em genes modificadores estão associados com a gravidade da FC. Método: Estudo de corte transversal, entre 2009 a 2010. Todos pacientes foram submetidos à análise das principais mutações no gene CFTR, presença dos polimorfismos (por meio de diferentes técnicas moleculares - "nested" PCR, digestão enzimática, PCR "multiplex", genotipagem em sequenciador automático e PCR ARMS) e características clínicas de gravidade da FC. Resultado: As mutações identificadas no gene CFTR foram associadas com variáveis que descrevem o início da doença, atuando como fator de proteção a clínica de maior gravidade [idade (p?0,0001), 1ª manifestação clínica (1MC) (p?0,0001), diagnóstico (p?0,0001), início dos sintomas pulmonares (ISP) (p?0,0001) e digestivos (ISD) (p?0,0001), 1ª bactéria Pseudomonas aeruginosa isolada (1PA) (p?0,0001), insuficiência pancreática (IP) (p?0,0001), P. aeruginosa mucóide (PAM) (p?0,0001), P. aeruginosa não mucóide (PANM) (p?0,0001)]. O gene ACE foi associado a 1MC (p:0,038), ao escore de Bhalla (EB) (p:0,049) e a presença de Bulkolderia cepacia (BC) (p:0,038). O polimorfismo GCLC129C>T foi associado à PAM (p:0,037). O polimorfismo GCLC350 foi associado a 1PA (p:0,049), Achromobacter xylosoxidans (p:0,044) e Staphylococcus aureus (SA) (p:0,037). O gene GSTM1 foi associado ao diagnóstico (p:0,014), índice de massa corpórea (IMC) (0,045), escore de Kanga (EK) (p:0,046), capacidade vital forçada [CVF(%)] (p:0,004) e volume expiratório forçado no primeiro segundo [VEF1(%)] (p:0,004). O gene GSTT1 foi associado ao ISD (p:0.033), osteoporose (p:0.021), EB (p:0,026), escore de Shwachman Kulczycki (SC) (p:0,009), CVF(%) (p:0,015), VEF1(%) (p:0,005), razão entre o VEF1 e a CVF [TIF(%)] (p:0,033), fluxo expiratório forçado entre 25 e 75% da CVF [FEF25- 75%(%)] (p:0,005) e PAM (p:0,038). O agrupamento genotípico dos genes GSTM1 e GSTT1 foi associado com o diagnóstico (p:0,014), BC (p:0,042) e SA (p:0,032). O gene GSTP1 foi associado com o ISP (p:0,026), IP (p:0,011), EB (p:0,015), EK (p:0,028), SC (p:0,039), CVF(%) (p:0,021), VEF1(%) (p:0,021), FEF25-75% (p:0,02) e 1PA (p:0,022). O alelo 1 para o polimorfismo de repetição AAT no gene NOS-1 foi associado com a idade (p:0,009) e com o ISD (p:0,001). O alelo 2 com ISP (p:0,031), ISD (p:0,001) e EB (p:0,046). O alelo 1 para o polimorfismo GT-1 no gene NOS-1 não apresentou associação com as variáveis clínicas. O alelo 2 apresentou associação com a saturação de oxigênio transcutânea (p:0,012), EB (p:0,013), CVF(%) (p:0,038), VEF1(%) (p:0,007), TIF(%) (p:0,036) e FEF25-75(%) (p:0,007). O alelo 1 para o polimorfismo de repetição GT-2 no gene NOS1 apresentou associação com a CVF(%) (p:0,032). O alelo 2 apresentou associação com o TIF(%) (p:0,009). O polimorfismo Arg>Gly no gene ADRB2R apresentou associação com a IP (p:0,009), EB (p:0,039), VEF1(%) (p:0,003), FEF25-75(%) (p:0,008), 1PA (p:0,012), PAM (p:0,023) e PANM (p:0,024). O polimorfismo Gln>Glu no gene ADRB2R apresentou associação com o EB (p:0,019) e TIF(%) (p:0,047). A resposta ao broncodilatador inalatório na prova de espirometria apresentou associação com os marcadores de gravidade clínica VEF1(%) (p:0,011) e FEF25-75(%) (p:0,019) apenas para o polimorfismo Arg>Gly no gene ADRB2R. Conclusão: Houve associação entre os polimorfismos analisados e as mutações no gene CFTR com a gravidade clínica da FC / Abstract: Introduction: Cystic fibrosis (CF) is clinically manifested in various forms of gravity, modulated by different genotypes and environment. While the classes of mutations in the CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) gene are well defined as modulators of severity in CF, polymorphisms remain controversial. Objective: To determine whether polymorphisms in modifier genes are associated with the severity in the CF. Method: Cross-sectional study, between 2009 to 2010. All patients were subjected to analysis of major mutations in CFTR gene, presence of polymorphisms (using different molecular techniques - Nested PCR, enzyme digestion, multiplex PCR, genotyping in automatic sequencer and ARMS PCR) and clinical severity in CF. Results: Mutations were identified in CFTR gene associated with variables that describe the onset of the disease, acting as a protective factor for the clinical severity [age (p?0.0001), first clinical manifestation (1CM) (p?0.0001), diagnosis (p?0.0001) , onset of pulmonary symptoms (OPS) (p?0.0001) and digestive (ODS) (p?0.0001), 1st isolated bacterium Pseudomonas aeruginosa (1PA) (p?0.0001), pancreatic insufficiency (PI) ( p?0.0001), P. aeruginosa mucoid (PAM) (p?0.0001) and P. aeruginosa no mucoid (PANM) (p?0.0001)]. ACE gene was associated with 1CM (p:0.038), Bhalla score (BS) (p:0.049) and presence of Bulkolderia cepacia (BC) (p:0.038). Polymorphism GCLC129C>T was associated with PAM (p:0.037). Polymorphism GCLC350 was associated with 1PA (p:0.049), Achromobacter xylosoxidans (p:0.044) and Staphylococcus aureus (SA) (p:0.037). GSTM1 gene was associated with diagnosis (p:0.014), body mass index (BMI) (0.045), Kanga score (KS) (p:0.046), forced vital capacity [FVC(%)] (p:0.004) and forced expiratory volume in one second [FEV1(%)] (p:0.004). GSTT1 gene was associated with ODS (p:0033), osteoporosis (p:0.021), BS (p:0.026), Shwachman-Kulczycki score (SS) (p:0.009), FVC(%) (p:0.015) , FEV1(%) (p:0.005), ratio between FEV1 and FVC [TIF(%)] (p:0.033), forced expiratory flow between 25 and 75% of FVC [FEF25-75%(%)] (p:0.005) and PAM (p:0.038). The genotypic groupings of genes GSTM1 and GSTT1 was associated with diagnosis (p:0.014), BC (p:0.042) and SA (p:0.032). GSTP1 gene was associated with the OPS (p:0.026), PI (p:0.011), BS (p:0.015), KS (p:0.028), SS (p:0.039), FVC(%) (p:0.021 ), FEV1(%) (p:0.021), FEF25-75% (p:0.02) and 1PA (p:0.022). Allele 1 for AAT repeat polymorphism in NOS-1 gene was associated with age (p:0.009) and ODS (p:0.001). Allele 2 with OPS (p:0.031), ODS (p:0.001) and BS (p:0.046). Allele 1 for repeat polymorphism GT-1 in NOS-1 gene was not associated with clinical variables. Allele 2 was associated with transcutaneous oxygen saturation (p:0.012), BS (p:0.013), FVC(%) (p:0.038), FEV1(%) (p:0.007), TIF(%) (p:0.036) and FEF25-75(%) (p:0.007). The allele 1 for repeat polymorphism GT-2 in NOS1 gene was associated with FVC(%) (p:0.032). The allele 2 was associated with the TIF(%) (p:0.009). Polymorphism Arg>Gly in ADRB2R gene was associated with PI (p:0.009), BS (p:0.039), FEV1(%) (p:0.003), FEF25-75(%) (p:0.008), 1PA ( p:0.012), PAM (p:0.023) and PANM (p:0.024). Polymorphism Gln>Glu in ADRB2R gene was associated with BS (p:0.019) and TIF(%) (p:0.047). The response to inhaled bronchodilator in the spirometry test presented association for the markers of clinical severity FEV1(%) (p:0.011) and FEF25-75(%) (p:0.019) only for the polymorphism Arg>Gly in ADRB2R gene. Conclusion: There was association between the polymorphisms studied and CFTR mutations with the clinical severity in CF / Mestrado / Saude da Criança e do Adolescente / Mestre em Saude da Criança e do Adolescente