Sample Preparation to Bioinformatics Analysis of DNA Methylation: Association Strategy for Obesity and Related Trait Studies

Autor(es):Mostrar menos -

Noronha, Natalia Yumi ; Rodrigues, Guilherme da Silva ; de Souza Pinhel, Marcela Augusta ; Cazier, Jean-Baptiste ; Watanabe, Ligia Moriguchi ; Menezes, Albert Nobre ; Bueno, Carlos Roberto ; Nicoletti, Carolina Ferreira ; Parenti de Oliveira, Bruno Affonso ; Schineider, Isabelle Mello ; Yonehara Noma, Isabella Harumi ; Dias Alcaras, Igor Caetano ; Barbosa, Fernando ; Nonino, Carla Barbosa

• Tipo de documento: Artigo Científico
• Fonte: JOVE-JOURNAL OF VISUALIZED EXPERIMENTS; v. N/A, n. 183, p. 20-pg., 2022-05-01.

Resumo

Obesity is directly connected to lifestyle and has been associated with DNA methylation changes that may cause alterations in the adipogenesis and lipid storage processes contributing to the development of the disease. We demonstrate a complete protocol from selection to epigenetic data analysis of patients with and without obesity. All steps from the protocol were tested and validated in a pilot study. 32 women participated in the study, in which 15 individuals were classified with obesity according to Body Mass Index (BMI) (45.1 +/- 5.4 kg/m(2)); and 17 individuals were classified without obesity according to BMI (22.6 +/- 1.8 kg/m(2)). In the group with obesity, 564 CpG sites related to fat mass were identified by linear regression analysis. The CpG sites were in the promoter regions. The differential analysis found 470 CpGs hypomethylated and 94 hypermethylated sites in individuals with obesity. The most hypomethylated enriched pathwayswere in the RUNX, WNT signaling, and response to hypoxia. The hypermethylated pathways were related to insulin secretion, glucagon signaling, and Ca2+. We conclude that the protocol effectively identified DNA methylation patterns and trait-related DNA methylation. These patterns could be associated with altered gene expression, affecting adipogenesis and lipid storage. Our results confirmed that an obesogenic lifestyle could promote epigenetic changes in human DNA. (AU)

• Processo FAPESP: 18/24069-3 - Do biomonitoramento ao reconhecimento de assinaturas do exposoma humano visando antecipar riscos para uma saúde contínua
• Beneficiário: Fernando Barbosa Júnior
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 14/16740-6 - Expressão de genes relacionados ao gasto energético e metabolismo de lipídios de indivíduos com obesidade grau III antes e após intervenções para perda de peso
• Beneficiário: Natália Yumi Noronha
• Modalidade de apoio: Bolsas no Brasil - Programa Capacitação - Treinamento Técnico