Proteomics reveals that the antifungal activity of fenbendazole against Cryptococcus neoformans requires protein kinases

Int J Antimicrob Agents. 2024 May;63(5):107157. doi: 10.1016/j.ijantimicag.2024.107157. Epub 2024 Mar 26.

Abstract

Cryptococcus neoformans is responsible for over 100 000 deaths annually, and the treatment of this fungal disease is expensive and not consistently effective. Unveiling new therapeutic avenues is crucial. Previous studies have suggested that the anthelmintic drug fenbendazole is an affordable and nontoxic candidate to combat cryptococcosis. However, its mechanism of anticryptococcal activity has been only superficially investigated. In this study, we examined the global cellular response of C. neoformans to fenbendazole using a proteomic approach (data are available via ProteomeXchange with identifier PXD047041). Fenbendazole treatment mostly impacted the abundance of proteins related to metabolic pathways, RNA processing, and intracellular traffic. Protein kinases, in particular, were significantly affected by fenbendazole treatment. Experimental validation of the proteomics data using a collection of C. neoformans mutants led to the identification of critical roles of five protein kinases in fenbendazole's antifungal activity. In fact, mutants lacking the expression of genes encoding Chk1, Tco2, Tco3, Bub1, and Sch9 kinases demonstrated greater resistance to fenbendazole compared to wild-type cells. In combination with the standard antifungal drug amphotericin B, fenbendazole reduced the cryptococcal burden in mice. These findings not only contribute to the elucidation of fenbendazole's mode of action but also support its use in combination therapy with amphotericin B. In conclusion, our data suggest that fenbendazole holds promise for further development as an anticryptococcal agent.

Keywords: Antifungal activity; Cryptococcus neoformans; Fenbendazole; Protein kinases; Proteomics.

MeSH terms

  • Amphotericin B / pharmacology
  • Animals
  • Antifungal Agents* / pharmacology
  • Cryptococcosis* / drug therapy
  • Cryptococcosis* / microbiology
  • Cryptococcus neoformans* / drug effects
  • Cryptococcus neoformans* / genetics
  • Disease Models, Animal
  • Drug Resistance, Fungal / genetics
  • Fenbendazole* / pharmacology
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Mice
  • Microbial Sensitivity Tests
  • Protein Kinases* / genetics
  • Protein Kinases* / metabolism
  • Proteomics*

Substances

  • Antifungal Agents
  • Fenbendazole
  • Protein Kinases
  • Amphotericin B
  • Fungal Proteins