Molecular characterization of the hexose transporter gene in benznidazole resistant and susceptible populations of Trypanosoma cruzi

Santos, Paula Fernandes dos; Ruiz, Jerônimo C.; Soares, Rodrigo Pedro; Moreira, Douglas S.; Rezende, Antônio Mauro; Folador, Edson Luiz; Oliveira, Guilherme; Romanha, Álvaro J.; Murta, Silvane Maria Fonseca

doi:10.1186/1756-3305-5-161

Cited by 5 publications

(3 citation statements)

References 46 publications

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“…Increased transcript abundance of 3-hydroxyisobutyryl-coenzyme a hydrolase-like protein in the early passages relative to the late is in consistency with the preparedness of the infection ready parasites to use beta-oxidation of fatty acids, as observed by others ( Alcolea et al, 2009 ) (Supplementary Data Sheet S4 ). As opposed to the earlier observations ( Moreno et al, 2014 ), level of tyrosine amino transferase presented a reverse trend, as it was slightly downregulated in the early passage, although protein expression may reveal different results as observed previously for Trypanosoma cruzi ( dos Santos et al, 2012 ). Purine acquisition and inter-conversion is an important determinant of environmental sensing and morphogenesis in Leishmania .…”

Section: Resultscontrasting

confidence: 99%

Genome Plasticity in Cultured Leishmania donovani: Comparison of Early and Late Passages

Sinha

Raghwan

et al. 2018

Front. Microbiol.

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Leishmania donovani possesses a complex heteroxenic life cycle where infective metacyclic promastigotes are pre-adapted to infect their host and cope up with intracellular stress. Exploiting the similarities between cultured and sandfly derived promastigotes, we used early and late passage cultured promastigotes to show specific changes at genome level which compromise pathogen fitness reflected in gene expression and infection studies. The pathogen loses virulence mostly via transcriptional and translational regulations and long-time cultivation makes them struggle to convert to virulent metacyclics. At the genomic level very subtle plasticity was observed between the early and the late passages mostly in defense-related, nutrient acquisition and signal transduction genes. Chromosome Copy number variation is seen in the early and late passages involving several genes that may be playing a role in pathogenicity. Our study highlights the importance of ABC transporters and calpain like cysteine proteases in parasite virulence in cultured promastigotes. Interestingly, these proteins are emerging as important patho-adaptive factors in clinical isolates of Leishmania. We found that the currently available genome of Leishmania in the NCBI database are from late passages. Our early passage genome can act as a reference for future studies on virulent isolates of Leishmania. The annotated leads from this study can be used for virulence surveillance and therapeutic studies in the Indian subcontinent.

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Section: Resultscontrasting

confidence: 99%

Genome Plasticity in Cultured Leishmania donovani: Comparison of Early and Late Passages

Sinha

Raghwan

et al. 2018

Front. Microbiol.

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“…Overexpression of enzymatic activity related to antioxidant response, like that of tryparedoxin peroxidase, ascorbate peroxidase, and iron-superoxide dismutase, has been detected in T. cruzi populations resistant to BZ (Nogueira et al, 2006(Nogueira et al, , 2009(Nogueira et al, , 2012. In addition, the hexose transporter activity was 40% lower in a BZ-resistant T. cruzi population than in the susceptible control (dos Santos et al, 2012). Differences in susceptibility to BZ and NFX between T. cruzi strains (Filardi and Brener, 1987;Toledo et al, 2004) and/or the genetic diversity of the host (Filardi and Brener, 1987) might explain, in part, the variations in the efficacies of antiparasitic drugs.…”

Section: Introductionmentioning

confidence: 96%

Molecular characterization of Cyclophilin (TcCyP19) in Trypanosoma cruzi populations susceptible and resistant to benznidazole

Rêgo

Duarte

Liarte

et al. 2015

Experimental Parasitology

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Cyclophilin (CyP), a peptidyl-prolyl cis/trans isomerase, is a key molecule with diverse biological functions that include roles in molecular chaperoning, stress response, immune modulation, and signal transduction. In this respect, CyP could serve as a potential drug target in disease-causing parasites. Previous studies employing proteomics techniques have shown that the TcCyP19 isoform was more abundant in a benznidazole (BZ)-resistant Trypanosoma cruzi population than in its susceptible counterpart. In this study, TcCyP19 has been characterized in BZ-susceptible and BZ-resistant T. cruzi populations. Phylogenetic analysis revealed a clear dichotomy between Cyphophilin A (CyPA) sequences from trypanosomatids and mammals. Sequencing analysis revealed that the amino acid sequences of TcCyP19 were identical among the T. cruzi samples analyzed. Southern blot analysis showed that TcCyP19 is a single-copy gene, located in chromosomal bands varying in size from 0.68 to 2.2 Mb, depending on the strain of T. cruzi. Northern blot and qPCR indicated that the levels of TcCyP19 mRNA were twofold higher in drug-resistant T. cruzi populations than in their drug-susceptible counterparts. Similarly, as determined by two-dimensional gel electrophoresis immunoblot, the expression of TcCyP19 protein was increased to the same degree in BZ-resistant T. cruzi populations. No differences in TcCyP19 mRNA and protein expression levels were observed between the susceptible and the naturally resistant T. cruzi strains analyzed. Taken together, these data indicate that cyclophilin TcCyP19 expression is up-regulated at both transcriptional and translational levels in T. cruzi populations that were in vitro-induced and in vivo-selected for resistance to BZ.

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“…In a previous study, the transcript level of the HT gene was observed to be more highly expressed in the BZ-resistant T. cruzi population. However, the authors observed a 40% reduction in transporter activity in this population, suggesting the existence of a regulatory mechanism at the protein activity level [ 52 ].…”

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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Molecular characterization of the hexose transporter gene in benznidazole resistant and susceptible populations of Trypanosoma cruzi

Cited by 5 publications

References 46 publications

Genome Plasticity in Cultured Leishmania donovani: Comparison of Early and Late Passages

Genome Plasticity in Cultured Leishmania donovani: Comparison of Early and Late Passages

Molecular characterization of Cyclophilin (TcCyP19) in Trypanosoma cruzi populations susceptible and resistant to benznidazole

Transcriptomic analysis of benznidazole-resistant and susceptible Trypanosoma cruzi populations

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