IL-2 gene expression in adipocytes from lupus patients in different nutritional status: in vitro study

• Grant number: 22/13266-8
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Effective date (Start): March 01, 2023
• Effective date (End): April 30, 2023
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Carolina Nicoletti Ferreira Fino
• Grantee: Amanda Alves Ribeiro
• Supervisor: Ana Belen Crujeiras Martinez
• Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Research place: Universidade de Santiago de Compostela, Vida (USC), Spain
• Associated to the scholarship: 21/09753-8 - Epigenetic effects of high fat diet in adipose tissue of animals with lupus, BP.MS

Abstract

Systemic lupus erythematosus (SLE) is associated with a complex immune dysregulation and its pathophysiology involves a multifactorial interaction between several genetic and epigenetic factors. The alteration in epigenetic patterns is proposed as an important contributing factor for the development of autoimmunity, and of the specific manifestations and treatment. The study aims to evaluated methylation levels of IL-2 gene in a culture of adipocytes grown with secretomas from lupus patients with normal weight or obesity. Cells will be incubated in 6-well cell culture plates (Iwaki, Tokyo, Japan) in 4 ml of serum-free medium that will be changed twice every two hours and again after 16h. Dulbeco's Modified Eagle's Medium (DMEM) without serum/fresh phenol red will be added and incubated for 24 hours. The cells will be incubated with human adipose tissue secretome for 24 h at concentrations of 1% and 5%. Treated with 11.3¼M folic acid, dissolved in 1M NaOH, and a methyl donor mixture consisting of folic acid, choline chloride and vitamin B12 after 48 h exposed to vehicle control in the first 24 h. at the end of the experiment RNA will be extracted. The gene expression analysis will be performed in triplicate by qPCR using the 7500 Fast Real PCR System (Applied Biosystems). In the statistical analysis, the Shapiro-Wilk tests, independent t test, Mann-Whitney U test, repeated measures ANOVA and/or Wilcoxon tests will be used. Interactions will be inferred through linear regressions and Pearson or Spearman correlation. Data analysis will be performed using the Statistical Package for Social Science(SPSS version 22.0 Inc. Chicago. IL). The level of significance will be set at p d 0.05. (AU)