microRNA expression in interaction of Cryptococcus neoformans and Glioblastome cells.

Ana Marisa Fusco-Almeida1
1.	Universidade Estadual Paulista, Júlio Mesquita Filho, UNESP, Arararaquara, SP, Brasil.

Cryptococcosis is an important systemic mycosis that mainly affects immunocompromised patients and is classified as the fungal infection with higher mortality among individuals with HIV. Although efficient, the treatment it is observed high number of cases of relapses, development of resistance and toxicity problems. In front of these problems, this study proposes the development of new therapies for the treatment of cryptococcosis, through of study microRNAs (miRNAs) involved in the interaction between yeast and human glioblastoma cells (U87-MG), in order to use them as regulators of infection, since this is a recently disclosed strategy in a variety of diseases. For this study, the interactions were performed by in vitro test with cellular culture and the infections were quantified by flow cytometry. Then the RNA of infected cells and normal cells were extracted, purified and quantified. The miRNA expression was evaluated by quantitative PCR using plate miRCURY LNATM Universal RT miRNA Ready-to-Use PCR for human cells. We observe that 10 miRNAs were over-expressed during the interaction of C. neoformans and U87-MG cells and it was noted that these miRNAs regulate the signaling pathway of TGF-β, MAP kinase (MAPK) and the receptor and matrix extracellular components (MEC) interaction. These data were compared with data from transcripts of the interaction and the GTPase-3 and COP-1 protein were selected as important in this process. Such proteins are associated to Rho-GT pathway Rho-GTPase which is essential for the yeast adhesion into host cells and showed increased expression in infection. Our results indicate that the inhibition of expression of miRNAs during fungus-cell interaction may be interesting to control fungal infection, because we believe that once inhibited, the yeast can reduce invasion of host cells, being the miRNAs interesting new therapeutic targets.

microRNA regulates antimicrobial responses expression during the infection of Cryptococcus neoformans