Dataset Information

Simvastatin Resistance of Leishmania amazonensis Induces Sterol Remodeling and Cross-Resistance to Sterol Pathway and Serine Protease Inhibitors.

ABSTRACT: The sterol biosynthesis pathway of Leishmania spp. is used as a pharmacological target; however, available information about the mechanisms of the regulation and remodeling of sterol-related genes is scarce. The present study investigated compensatory mechanisms of the sterol biosynthesis pathway using an inhibitor of HMG-CoA reductase (simvastatin) and by developing drug-resistant parasites to evaluate the impact on sterol remodeling, cross-resistance, and gene expression. Simvastatin-resistant L. amazonensis parasites (LaSimR) underwent reprogramming of sterol metabolism manifested as an increase in cholestane- and stigmastane-based sterols and a decrease in ergostane-based sterols. The levels of the transcripts of sterol 24-C-methyltransferase (SMT), sterol C14-α-demethylase (C14DM), and protease subtilisin (SUB) were increased in LaSimR. LaSimR was cross-resistance to ketoconazole (a C14DM inhibitor) and remained sensitive to terbinafine (an inhibitor of squalene monooxygenase). Sensitivity of the LaSimR mutant to other antileishmanial drugs unrelated to the sterol biosynthesis pathway, such as trivalent antimony and pentamidine, was similar to that of the wild-type strain; however, LaSimR was cross-resistant to miltefosine, general serine protease inhibitor N-p-tosyl-l-phenylalanine chloromethyl ketone (TPCK), subtilisin-specific inhibitor 4-[(diethylamino)methyl]-N-[2-(2-methoxyphenyl)ethyl]-N-(3R)-3-pyrrolidinyl-benzamide dihydrochloride (PF-429242), and tunicamycin. The findings on the regulation of the sterol pathway can support the development of drugs and protease inhibitors targeting this route in parasites.

SUBMITTER: Fujii TTS

PROVIDER: S-EPMC8877030 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature

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Simvastatin Resistance of <i>Leishmania amazonensis</i> Induces Sterol Remodeling and Cross-Resistance to Sterol Pathway and Serine Protease Inhibitors.

Fujii Thais Tenorio Soares TTS Gomes Pollyanna Stephanie PS do Monte-Neto Rubens Lima RL de Oliveira Gomes Daniel Claudio DC Ouellette Marc M Torres-Santos Eduardo Caio EC Andrade-Neto Valter Viana VV de Matos Guedes Herbert Leonel HL

Microorganisms 20220209 2

The sterol biosynthesis pathway of <i>Leishmania</i> spp. is used as a pharmacological target; however, available information about the mechanisms of the regulation and remodeling of sterol-related genes is scarce. The present study investigated compensatory mechanisms of the sterol biosynthesis pathway using an inhibitor of HMG-CoA reductase (simvastatin) and by developing drug-resistant parasites to evaluate the impact on sterol remodeling, cross-resistance, and gene expression. Simvastatin-re ...[more]

PMID: 35208853

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Project description:BackgroundLeishmaniases are neglected tropical diseases with great clinical and epidemiological importance. The current chemotherapy available for the treatment of leishmaniasis presents several problems, such as adverse effects, toxicity, long treatment time, and parasite resistance. The discovery of new therapeutic alternatives is extremely essential, and the discovery of cellular targets is a tool that helps in the development of new drugs. Serine proteases emerge as important virulence factors in the Leishmania genus, as they participate in important processes involved in their infectivity, virulence, and survival. In this work, we evaluated the leishmanicidal effect of different serine protease inhibitors (Benzamidine, PF-429242, PMSF, TLCK, and TPCK). Additionally, we determined the implication of pretreatment with these inhibitors on the entry and survival of parasites within macrophages, as well as the conversion of promastigotes into amastigotes, to discover the importance of serine proteases in the establishment of infection and, consequently, as targets for new drugs for Leishmania.ResultsIn general, the inhibitors had low toxicity in host macrophages, and three showed some effect in promastigote and amastigote forms of L. amazonensis (PF-429242, TLCK, and TPCK). Using a short incubation interval, we pretreated L. amazonensis promastigotes with these five compounds before in vitro infection. Pretreatment with PF-429242, TLCK, and TPCK considerably compromised the survival of these parasites inside host macrophages, without altering the entry of promastigotes into these cells and differentiation into amastigotes. In addition, treatment with PF-429242 and TPCK was able to reduce the serine proteases' enzymatic activity using subtilisin substrate on L. amazonensis promastigote lysate.ConclusionsThis work highlights the importance of serine proteases in L. amazonensis as a possible target for new therapeutic alternatives in Leishmania spp.

Project description:Leishmaniasis encompass a group of infectious parasitic diseases occurring in 97 endemic countries where over one billion people live in areas at risk of infection. It is in the World Health Organization list of neglected diseases and it is considered a serious public health problem, with more than 20,000 deaths a year and high morbidity. Infection by protozoa from the genus Leishmania can cause several forms of the disease, which may vary from a self-healing ulcer to fatal visceral infection. Leishmania species, as well as host immune response and genetics can modulate the course of the disease. Leishmania sp are obligatory intracellular parasites that have macrophages as their main host cell. Depending on the activation phenotype, these cells may have distinct roles in disease development, acting in parasite control or proliferation. Therefore, the purpose of this work was to analyze Leishmania amazonensis infection in primary macrophage cells obtained from mice with two distinct genetic backgrounds, ie. different susceptibility to the infection; evaluating the cause for that difference. After infection, peritoneal macrophages from the resistant C3H/He strain presented lower parasite load when compared to susceptible BALB/c macrophages. The same was also true when cells received a Th2 stimulus after infection, but the difference was abrogated under Th1 stimulus. Nitric oxide production and arginase activity was different between the strains under Th1 or Th2 stimulus, respectively, but iNOS inhibition was unable to suppress C3H/He resistance. Hydrogen peroxide production was also higher in C3H/He than BALB/c under Th1 stimulus, but it could not account for differences in susceptibility. These results led us to conclude that, although they have an important role in parasite control, neither NO nor H2O2 production can explain C3H/He resistance to infection. Other studies are needed to uncover different mechanisms of resistance/susceptibility to L. amazonensis.

Dataset Information

Simvastatin Resistance of Leishmania amazonensis Induces Sterol Remodeling and Cross-Resistance to Sterol Pathway and Serine Protease Inhibitors.

Publications

Simvastatin Resistance of <i>Leishmania amazonensis</i> Induces Sterol Remodeling and Cross-Resistance to Sterol Pathway and Serine Protease Inhibitors.

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