Author(s): Araújo, Daniela ; Azevedo, N. ; Barbosa, Ana ; Almeida, Carina ; Rodrigues, M. Elisa ; Henriques, Mariana ; Silva, Sónia Carina
Date: 2019
Persistent ID: http://hdl.handle.net/1822/61935
Origin: RepositóriUM - Universidade do Minho
Project/scholarship: info:eu-repo/grantAgreement/FCT/5876/147337/PT;
Subject(s): Candidiasis; Filamentation; Nucleic Acid Mimics; 2-MethylRNA modification; 2′-OMethylRNA modification; Science & Technology
Antisense oligomers and their analogues have been successfully utilized to silence gene expression for the treatment of many human diseases; however the control of yeast´s virulence determinants has never been exploited before. In this sense, this work is based on the key hypothesis that if a pathogens genetic sequence is a determinant of virulence, it will be possible to synthesize a nucleic acid mimic based on antisense therapy (AST) that will bind to the mRNA produced, blocking its translation into protein and consequently reducing the pathogen virulent phenotype. EFG1 is an important determinant of virulence that is involved in regulation of Candida albicans switch from yeast to filamentous form. Thus, our main goal was to design and synthesize an antisense oligonucleotide (ASO) targeting the EFG1 mRNA and to validate its in vitro applicability. The results show that the anti-EFG1 2-OMethylRNA (2OMe) oligomer was able to significantly reduce the levels of EFG1 gene expression and of Efg1p protein translation (both approximately 60%), as well as effectively prevent filamentation of C. albicans cells (by 80%). Moreover, it was verified that anti-EFG1 2OMe keep the efficacy in different simulated human body fluids. Undeniably, this work provides potentially valuable information for future research into the management of Candida infections, regarding the development of a credible and alternative method to control C. albicans infections, based on antisense therapy methodology.
This study was supported by the Portuguese Foundation for Science and Technology (FCT), under the scope of the strategic funding of the UID/BIO/04469 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684) and the BioTecNorte operation (NORTE-01-0145-FEDER-000004), funded by the European Regional Development Fund under the scope of a Norte 2020 – Programa Operacional Regional do Norte and Daniela Eira Araújo (SFRH/BD/121417/2016) PhD grant. The authors also acknowledge the project funding by the “02/SAICT/2017 – Projetos de Investigação Científica e Desenvolvimento Tecnológico (IC&DT; POCI-01-0145-FEDER-028893).” The mass spectrometry technique was performed at the Proteomics i3S Scientific Platform with the assistance of Hugo Osório. This work had support from the Portuguese Mass Spectrometry Network, integrated in the National Roadmap of Research Infrastructures of Strategic Relevance (ROTEIRO/0028/2013 and LISBOA-01-0145-FEDER-022125).
info:eu-repo/semantics/publishedVersion
