Caracterização do perfil clínico-epidemiológico de uma população com hanseníase na Bahia e avaliação de microRNAs (miRNAs) ligados à via dos TLRs e sua correlação com genes de morte celular em biópsias de pacientes com hanseníase

Abstract

Introduction: Leprosy is a chronic infectious disease caused by Mycobacterium leprae. About 30-40% of patients develop sudden and acute inflammatory episodes, the so-called leprosy reactions. Previous studies have shown a series of miRNAs associated with leprosy phenotypes, evidencing the role of this epigenetic mechanism in the pathogenesis of the disease. Objective: This study aimed to evaluate the correlation of miRNAs previously associated with leprosy with genes linked to apoptosis and autophagy in lesions of patients with leprosy, compare the expression pattern of genes in biopsies of patients with and without reactions and describe the clinical profile and epidemiological analysis of patients with leprosy treated at reference centers in the state of Bahia, HUPES-AMN and ICOM. Methods: Biopsies of 28 patients with and without leprosy reactions, with different clinical forms of leprosy were recruited. For analysis purposes, these patients were allocated according to the WHO classification as paucibacillary (PB) or multibacillary (MB), and separated into patients with or without reactions. RNA samples contained in our biorepository were used, extracted by the method using TRIzol® Reagent (Ambion®). Obtaining complementary DNA (cDNA) was performed using the commercially available High Capacity kit (ThermoFisher). The genes were chosen because they are important in cell death processes and are regulated by the miRNAs previously analyzed by the group. Gene expression by real-time qPCR method by TaqMan® was performed and the data analyzed by comparing the threshold cycle (Ct) between groups of samples. Correlation analysis between genes and miRNAs was performed using the Spearman Correlation test, using GraphPad Prism8. Differences were considered significant when the p-value was below 0.05 (p ˂ 0.05). The patients' clinical-epidemiological database was analyzed using the R program for Windows, version 4.2.3 and the data presented in tables. Results: The expression of the genes ATG12, TNFRSF10A, PARK2, BCL2, CFLAR and STX7 was evaluated by qPCR and the expression between patients without reaction (PB + MB) and patients with reaction (RR and ENH) was compared and there was no statistically significant association in this comparison. Our results showed a correlation between miRNAs and ATG12 (miR-125a-3p), TNFRSF10A (miR-146b-5p), PARK2, CFLAR and STX7 (miR-132-5p) genes. In the epidemiological analysis, we observed that the MB form is more prevalent in males and that disease forms within this spectrum are more prone to the development of reactions, as well as affecting patients with lower educational and economic levels. Conclusion: We reinforce a role for these miRNAs in the pathogenesis of leprosy, influencing mechanisms such as apoptosis and autophagy in the skin; The MB form of the disease occurs within a social context of greater social vulnerability compared to PB patients.