Mitochondrial dysfunction on Leishmania (Leishmania) amazonensis induced by ketoconazole: insights into drug mode of action

Abstract

BACKGROUND Leishmania parasites cause leishmaniasis that range from self-limiting cutaneous lesions to more serious forms of the disease. The search for potential drug targets focusing on biochemical and metabolic pathways revealed the sterol biosynthesis inhibitors (SBIs) as a promising approach. In this class of inhibitors is found ketoconazole, a classical inhibitor of 14α-methysterol 14-demethylase. OBJECTIVE The present study aimed to better understand the biological response of Leishmania (Leishmania) amazonensis promastigotes at the cellular level after ketoconazole treatment. METHODS Herein, techniques, such as fluorimetry, flow cytometry, fluorescence microscopy, electron and scanning microscopy were used to investigate the cellular structures and to identify organelles affected by ketoconazole treatment. FINDINGS The study demonstrated, for the first time, the effect of ketoconazole on mitochondrion functioning and its probable relationship to cell cycle and death on L. (L.) amazonensis promastigotes (IFLA/BR/67/PH8 strain). MAIN CONCLUSIONS Ketoconazole-induced mitochondrial damages led to hyperpolarisation of this single organelle and autophagic vacuoles formation, as a parasite survival strategy. These damages did not reflect directly on the parasite cell cycle, but drove the parasites to death, making them susceptible to ketoconazole treatment in in vitro models.

PUF3 RNA binding protein of Trypanosoma cruzi regulates mitochondrial morphology and function

“…PUF3 overexpressor T. cruzi parasites exhibited severe ultrastructural changes, such as mitochondrial swelling. Scientific evidence reveals that different types of compounds can cause these changes and alterations in mitochondrial ultrastructure in trypanosomatids, such as ketoconazole, ruthenium, acriflavine, antricide, pentamidine, puromycin, suramin and mapharside [ [45] , [46] , [47] , [48] ], showing that these parasites are under much stress which explains all the genetic and biological changes. Also, these changes have been associated with the interference of mitochondrial activity, shown by changes in the ΔΨm [ 45 ].…”

“…Scientific evidence reveals that different types of compounds can cause these changes and alterations in mitochondrial ultrastructure in trypanosomatids, such as ketoconazole, ruthenium, acriflavine, antricide, pentamidine, puromycin, suramin and mapharside [ [45] , [46] , [47] , [48] ], showing that these parasites are under much stress which explains all the genetic and biological changes. Also, these changes have been associated with the interference of mitochondrial activity, shown by changes in the ΔΨm [ 45 ]. However, the absence of ultrastructural evidence in the knockout parasites does not exclude effects on other aspects of physiology and metabolism that may affect other organelles.…”

Hypothesis-generating proteome perturbation to identify NEU-4438 and acoziborole modes of action in the African Trypanosome

“…To identify physiologic targets of a drug, investigators need information on (i) the list of drug-binding proteins, and (ii) a catalog of modes of action of a drug ( Figure 10 ). “Modes of action” correspond to those molecular pathways whose inhibition by a drug compromises cell viability ( Bernacchia et al., 2022 ; Lytton et al., 1993 ; Ma et al., 2022 ; Nunes et al., 2022 ; Pandit et al., 2022 ; Schafer and Wenzel, 2020 ) ( Figure 10 ). Two experimental strategies can be used to obtain mode of action data.…”

Monomethylsulochrin isolated from biomass extract of Aspergillus sp. against Leishmania amazonensis: In vitro biological evaluation and molecular docking

“…The antileishmanial activity of flavonoid epigallocatechin-3-gallate (EGCG) against L. amazonensis promastigotes is associated with mitochondrial dysfunction causing parasite death ( Inacio et al., 2012 ). On the same way, ketoconazole ( Nunes et al., 2022 ), carajurin ( Silva-Silva et al., 2022 ), and dihydroartemisinin ( Grazzia et al., 2021 ) had their antileishmanial activity associated with mitochondrial dysfunction that interfered in Leishmania survival.…”