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In vitro functional characterization of microRNAs in toxoplasmosis

Grant number: 22/01475-1
Support type:Scholarships in Brazil - Master
Effective date (Start): September 01, 2022
Effective date (End): August 31, 2024
Field of knowledge:Health Sciences - Collective Health - Public Health
Principal researcher:Cristina da Silva Meira Strejevitch
Grantee:Tamires Santos de Arruda
Home Institution: Instituto Adolfo Lutz (IAL). Coordenadoria de Controle de Doenças (CCD). Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

Recent studies show that microRNAs (miRNAs), small molecules that play an important role in the regulation of gene expression in eukaryotic cells, are capable of negatively regulating the processing, stability and translation of several target messenger RNAs for the production of cytokines. However, the regulatory mechanisms involved during Toxoplasma gondii infection still remain unknown. Previous findings by our group on the characterization of the immune response in patients with symptomatic toxoplasmosis (ocular, cerebral and congenital) identified potential miRNAs that could be involved in the regulation of the production of important cytokines for infection control. Such miRNAs are undergoing in silico analysis and their predicted targets are being determined. Thus, the present project consists of carrying out the in vitro functional characterization of these miRNAs through overexpression and silencing assays in order to validate the findings obtained from the in silico analysis and confirm the potential effects of miRNAs on their targets. Furthermore, we intend to measure its effects on cellular processes and its impact on the production of cytokines important for infection control. The experiments will be conducted by infection of human macrophages of the THP-1 strain by T. gondii and gene expression analyzes will be performed by real-time PCR. We believe that the investigation of the performance of these molecules in the regulation of the immune system is timely and relevant. The correlation of miRNA and the profile of the cytokine gene expression response broadens the understanding of the molecular relationship established between host and parasite during infection. These assays are extremely important for the continuity of a larger project in which we intend to study the role of miRNAs in symptomatic toxoplasmosis. In addition, these findings may provide information about this important mechanism of gene silencing in toxoplasmosis, in addition to the learning and scientific enrichment that this project will provide.

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