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Analysis of the CREB3 regulatory factor importance on the replication of Zika virus in trophoblasts.

Grant number: 23/05503-2
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): July 01, 2023
Effective date (End): December 31, 2023
Field of knowledge:Biological Sciences - Microbiology
Principal Investigator:Eurico de Arruda Neto
Grantee:Matheus Henrique Pereira da Silva
Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated research grant:19/26119-0 - Emerging and re-emerging viruses: biology, pathogenesis and prospection, AP.TEM

Abstract

The Zika virus (ZIKV) is an important arbovirus transmitted by mosquitoes of the Aedes genus. It was isolated for the first time in 1947 in the Zika forest, located in Uganda. In 2015, the virus arrived in Brazil causing several outbreaks across the country and was quickly associated with the occurrence of congenital syndrome in fetuses gestated by infected women. Transplacental transmission was proved by clinical evidence and in vivo and in vitro experiments, including phenotyping of ZIKV-infected cells. Trophoblasts are cell types recognized for their resistance to viral infections, but they are susceptible and permissive to ZIKV. In addition, autophagy, a cellular process that can perform pro or antiviral functions, has been described as an important factor that favors virus replication in these cells. Since there are studies showing that the CREB3 regulatory protein (CREBRF) is an important repressor of autophagy, our research group carried out the silencing of CREBRF in cells of the JEG-3 lineage and observed an increase in the replication of ZIKV genome. Therefore, in the present project, we aim to investigate the expression of viral proteins and the generation of infectious progeny in JEG-3 cells silenced or overexpressing CREBRF by using Western Blot and titration by plaque assay. In addition, primary cultures of purified human placental cytotrophoblasts will be prepared. These cultures will be infected in vitro to evaluate CREBRF expression at the transcriptional and post-transcriptional level to understand whether ZIKV infection affects the expression of this protein as a way to induce autophagy and favor viral replication.

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