Transcriptome Profile of the Response of Paracoccidioides spp. to a Camphene Thiosemicarbazide Derivative.

Lívia Do Carmo Silva,Narcimário Pereira Coelho,Célia Maria De Almeida Soares,Symone Vitoriano Da Conceição Castro,Juliana Alves Parente-Rocha,Cecília Maria Alves De Oliveira,Juan G Mcewen Ochoa,Orville Hernández Ruiz,Maristela Pereira,Alexandre Melo Bailão,Ludmila Bringel Pires,Diana Patrícia Tamayo Ossa,Cleuza Conceição Da Silva

doi:10.1371/journal.pone.0130703

Abstract

Paracoccidioidomycosis (PCM) is a systemic granulomatous human mycosis caused by fungi of the genus Paracoccidioides, which is geographically restricted to Latin America. Inhalation of spores, the infectious particles of the fungus, is a common route of infection. The PCM treatment of choice is azoles such as itraconazole, but sulfonamides and amphotericin B are used in some cases despite their toxicity to mammalian cells. The current availability of treatments highlights the need to identify and characterize novel targets for antifungal treatment of PCM as well as the need to search for new antifungal compounds obtained from natural sources or by chemical synthesis. To this end, we evaluated the antifungal activity of a camphene thiosemicarbazide derivative (TSC-C) compound on Paracoccidioides yeast. To determine the response of Paracoccidioides spp. to TSC-C, we analyzed the transcriptional profile of the fungus after 8 h of contact with the compound. The results demonstrate that Paracoccidioides lutzii induced the expression of genes related to metabolism; cell cycle and DNA processing; biogenesis of cellular components; cell transduction/signal; cell rescue, defense and virulence; cellular transport, transport facilities and transport routes; energy; protein synthesis; protein fate; transcription; and other proteins without classification. Additionally, we observed intensely inhibited genes related to protein synthesis. Analysis by fluorescence microscopy and flow cytometry revealed that the compound induced the production of reactive oxygen species. Using an isolate with down-regulated SOD1 gene expression (SOD1-aRNA), we sought to determine the function of this gene in the defense of Paracoccidioides yeast cells against the compound. Mutant cells were more susceptible to TSC-C, demonstrating the importance of this gene in response to the compound. The results presented herein suggest that TSC-C is a promising candidate for PCM treatment.

Highlights

Paracoccidioidomycosis (PCM) is a systemic mycosis geographically restricted to Latin America caused by thermodimorphic fungi of the genus Paracoccidioides
tiosemicarbazide camphene derivative (TSC-C) at a concentration of 79 μM inhibited the cellular growth by 50% and became the IC50 value of TSC-C for Paracoccidioides yeast
Because TSC-C induces the production of Reactive oxygen species (ROS), we evaluated the mitochondrial membrane integrity by estimating the electric potential (ΔCm) with fluorescence in yeast grown in the presence of TSC-C for 4, 8 and 12 h

Summary

Introduction

Paracoccidioidomycosis (PCM) is a systemic mycosis geographically restricted to Latin America caused by thermodimorphic fungi of the genus Paracoccidioides. The fungi usually infect the host through the respiratory tract by inhalation of conidia, which are the infectious propagules found in the environment In the lungs, these propagules differentiate into the pathogenic form in a temperature-dependent manner, corresponding to the yeast phase of the fungus, and spreads to other organs through lymphohematogenous dissemination. These propagules differentiate into the pathogenic form in a temperature-dependent manner, corresponding to the yeast phase of the fungus, and spreads to other organs through lymphohematogenous dissemination Because this mycosis affects mainly rural males of working age between the ages of 30 and 50 years, the disease has socioeconomic repercussions due to its potential to debilitate. The need to research and develop new therapeutic approaches is increasingly evident With this aim, our group has invested effort into identifying and characterizing novel targets for antifungal drugs against Paracoccidioides spp. Our group has invested effort into identifying and characterizing novel targets for antifungal drugs against Paracoccidioides spp. [8,9,10,11,12,13,14,15,16] and searching for new antifungal compounds obtained from natural sources or their synthetic derivatives [17,18,19]

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