Overexpression of eukaryotic initiation factor 5A (eIF5A) affects susceptibility to benznidazole in Trypanosoma cruzi populations

Moreira, Douglas de Souza; Duarte, Ana Paula; Pais, Fabiano Sviatopolk-Mirsky; Silva-Pereira, Rosiane Aparecida da; Romanha, Álvaro J.; Schenkman, Sérgio; Murta, Silvane Maria Fonseca

doi:10.1590/0074-02760180162

Cited by 5 publications

(5 citation statements)

References 28 publications

(30 reference statements)

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“…The metabolites of Bz can also affect the metabolism of the protozoan, enhancing its trypanocidal effect without acting in a lytic way. (31,32) Together, our results suggest that drug screening against GFP-expressing T. cruzi epimastigotes should be carefully evaluated to avoid potential errors. Although epimastigotes (insect forms) are easily cultured in laboratory conditions, these forms are not found in the mammalian hosts.…”

Section: Discussionmentioning

confidence: 81%

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Improving in vitro screening compounds anti-Trypanosoma cruzi by GFP-expressing parasites

Delvoss,

Inoue,

da Silva

et al. 2024

Mem. Inst. Oswaldo Cruz

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BACKGROUND Conventional microscopic counting is a widely utilised method for evaluating the trypanocidal effects of drugs on intracellular amastigotes. This is a low-cost approach, but it is time-consuming and reliant on the expertise of the microscopist. So, there is a pressing need for developing technologies to enhance the efficiency of low-cost anti- Trypanosoma cruzi drug screening. OBJECTIVES In our laboratory, we aimed to expedite the screening of anti- T. cruzi drugs by implementing a fluorescent method that correlates emitted fluorescence from green fluorescent protein (GFP)-expressing T. cruzi (Tc-GFP) with cellular viability. METHODS Epimastigotes (Y strain) were transfected with the pROCKGFPNeo plasmid, resulting in robust and sustained GFP expression across epimastigotes, trypomastigotes, and intracellular amastigotes. Tc-GFP epimastigotes and intracellular amastigotes were exposed to a serial dilution of benznidazole (Bz). Cell viability was assessed through a combination of microscopic counting, MTT, and fluorimetry. FINDINGS The fluorescence data indicated an underestimation of the activity of Bz against epimastigotes (IC 50 75 µM x 14 µM). Conversely, for intracellular GFP-amastigotes, both fluorimetry and microscopy yielded identical IC 50 values. Factors influencing the fluorimetry approach are discussed. MAIN CONCLUSIONS Our proposed fluorometric assessment is effective and can serve as a viable substitute for the time-consuming microscopic counting of intracellular amastigotes.

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Section: Discussionmentioning

confidence: 81%

“…The metabolites of Bz can also affect the metabolism of the protozoan, enhancing its trypanocidal effect without acting in a lytic way. 31 , 32 …”

Section: Discussionmentioning

confidence: 99%

Improving in vitro screening compounds anti-Trypanosoma cruzi by GFP-expressing parasites

Delvoss,

Inoue,

da Silva

et al. 2024

Mem. Inst. Oswaldo Cruz

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“…Previous studies have also shown that eIF5A protein levels were twofold lower in BZ-resistant T. cruzi populations. In addition, mutant parasites that overexpress eIF5A are three to five times more susceptible to BZ [ 56 ]. Among the categories enriched in the upregulated transcripts in the BZ-resistant T. cruzi population, translation, aromatic amino acid metabolism and arginine metabolic processes were observed among the categories enriched in the upregulated transcripts in the BZ-resistant T. cruzi population.…”

Section: Discussionmentioning

confidence: 99%

Transcriptomic analysis of benznidazole-resistant and susceptible Trypanosoma cruzi populations

Lima

Gonçalves²,

Reis-Cunha

et al. 2023

Parasites Vectors

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Background Chagas disease (CD), caused by the parasite Trypanosoma cruzi, is a serious public health concern in Latin America. Nifurtimox and benznidazole (BZ), the only two drugs currently approved for the treatment of CD, have very low efficacies in the chronic phase of the disease and several toxic side effects. Trypanosoma cruzi strains that are naturally resistant to both drugs have been reported. We performed a comparative transcriptomic analysis of wild-type and BZ-resistant T. cruzi populations using high-throughput RNA sequencing to elucidate the metabolic pathways related to clinical drug resistance and identify promising molecular targets for the development of new drugs for treating CD. Methods All complementary DNA (cDNA) libraries were constructed from the epimastigote forms of each line, sequenced and analysed using the Prinseq and Trimmomatic tools for the quality analysis, STAR as the aligner for mapping the reads against the reference genome (T. cruzi Dm28c—2018), the Bioconductor package EdgeR for statistical analysis of differential expression and the Python-based library GOATools for the functional enrichment analysis. Results The analytical pipeline with an adjusted P-value of < 0.05 and fold-change > 1.5 identified 1819 transcripts that were differentially expressed (DE) between wild-type and BZ-resistant T. cruzi populations. Of these, 1522 (83.7%) presented functional annotations and 297 (16.2%) were assigned as hypothetical proteins. In total, 1067 transcripts were upregulated and 752 were downregulated in the BZ-resistant T. cruzi population. Functional enrichment analysis of the DE transcripts identified 10 and 111 functional categories enriched for the up- and downregulated transcripts, respectively. Through functional analysis we identified several biological processes potentially associated with the BZ-resistant phenotype: cellular amino acid metabolic processes, translation, proteolysis, protein phosphorylation, RNA modification, DNA repair, generation of precursor metabolites and energy, oxidation–reduction processes, protein folding, purine nucleotide metabolic processes and lipid biosynthetic processes. Conclusions The transcriptomic profile of T. cruzi revealed a robust set of genes from different metabolic pathways associated with the BZ-resistant phenotype, proving that T. cruzi resistance mechanisms are multifactorial and complex. Biological processes associated with parasite drug resistance include antioxidant defenses and RNA processing. The identified transcripts, such as ascorbate peroxidase (APX) and iron superoxide dismutase (Fe-SOD), provide important information on the resistant phenotype. These DE transcripts can be further evaluated as molecular targets for new drugs against CD. Graphical Abstract

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“…N-(2-amino2-hyroxymethyl) lysine, also called hypusine, is found in two eukaryotic proteins, eukaryotic translation initiation factor (eIF)-5A1 and eIF5A2 [ 121 ]. eIF5A is responsible for cell cycle regulation, apoptosis, translation, elongation, and termination [ 122 ]. The post-translational modification leads to the binding of hypusine with lysine residues of eIF5A proteins by the transfer of 4-aminobutyl moiety from spermidine to the lysine chain, which is aided by the enzymes deoxyhypusine synthase (DHPS) and deoxyhypusine hydroxylase (DOHH), also known as hypusination [ 123 ].…”

Section: Leishmaniasis and Chagas Diseasementioning

confidence: 99%

The Potential Use of Peptides in the Fight against Chagas Disease and Leishmaniasis

Berhe,

Kumar Cinthakunta Sridhar,

Zerihun

et al. 2024

Pharmaceutics

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Chagas disease and leishmaniasis are both neglected tropical diseases that affect millions of people around the world. Leishmaniasis is currently the second most widespread vector-borne parasitic disease after malaria. The World Health Organization records approximately 0.7–1 million newly diagnosed leishmaniasis cases each year, resulting in approximately 20,000–30,000 deaths. Also, 25 million people worldwide are at risk of Chagas disease and an estimated 6 million people are infected with Trypanosoma cruzi. Pentavalent antimonials, amphotericin B, miltefosine, paromomycin, and pentamidine are currently used to treat leishmaniasis. Also, nifurtimox and benznidazole are two drugs currently used to treat Chagas disease. These drugs are associated with toxicity problems such as nephrotoxicity and cardiotoxicity, in addition to resistance problems. As a result, the discovery of novel therapeutic agents has emerged as a top priority and a promising alternative. Overall, there is a need for new and effective treatments for Chagas disease and leishmaniasis, as the current drugs have significant limitations. Peptide-based drugs are attractive due to their high selectiveness, effectiveness, low toxicity, and ease of production. This paper reviews the potential use of peptides in the treatment of Chagas disease and leishmaniasis. Several studies have demonstrated that peptides are effective against Chagas disease and leishmaniasis, suggesting their use in drug therapy for these diseases. Overall, peptides have the potential to be effective therapeutic agents against Chagas disease and leishmaniasis, but more research is needed to fully investigate their potential.

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Overexpression of eukaryotic initiation factor 5A (eIF5A) affects susceptibility to benznidazole in Trypanosoma cruzi populations

Cited by 5 publications

References 28 publications

Improving in vitro screening compounds anti-Trypanosoma cruzi by GFP-expressing parasites

Improving in vitro screening compounds anti-Trypanosoma cruzi by GFP-expressing parasites

Transcriptomic analysis of benznidazole-resistant and susceptible Trypanosoma cruzi populations

The Potential Use of Peptides in the Fight against Chagas Disease and Leishmaniasis

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