Drug resistance analysis by next generation sequencing in Leishmania

Leprohon, Philippe; Fernández-Prada, Christopher; Gazanion, Élodie; Monte-Neto, Rubens L.; Ouellette, Marc

doi:10.1016/j.ijpddr.2014.09.005

Cited by 71 publications

(57 citation statements)

References 139 publications

(204 reference statements)

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“…However, the role of the LABCG2 transporter in drug resistance has not yet been elucidated. Modulation of gene expression through gene amplification and gene deletion by homologous recombination is a common mechanism of drug resistance in Leishmania strains derived from both the laboratory and the field thanks to the plasticity of the Leishmania genome (25)(26)(27). We are therefore interested in determining whether the overexpression of LABCG2 in a L. major line (data not shown) could confer drug resistance.…”

Section: Resultsmentioning

confidence: 99%

The LABCG2 Transporter from the Protozoan Parasite Leishmania Is Involved in Antimony Resistance

Perea

Manzano

Castanys

et al. 2016

Antimicrob Agents Chemother

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Treatment for leishmaniasis, which is caused by Leishmania protozoan parasites, currently relies on a reduced arsenal of drugs. However, the significant increase in the incidence of drug therapeutic failure and the growing resistance to first-line drugs like antimonials in some areas of Northern India and Nepal limit the control of this parasitic disease. Understanding the molecular mechanisms of resistance in Leishmania is now a matter of urgency to optimize drugs used and to identify novel drug targets to block or reverse resistant mechanisms. Some members of the family of ATP-binding cassette (ABC) transporters in Leishmania have been associated with drug resistance. In this study, we have focused our interest to characterize LABCG2's involvement in drug resistance in Leishmania. Leishmania major parasites overexpressing the ABC protein transporter LABCG2 were generated in order to assess how LABCG2 is involved in drug resistance. Assays of susceptibility to different leishmanicidal agents were carried out. Analysis of the drug resistance profile revealed that Leishmania parasites overexpressing LABCG2 were resistant to antimony, as they demonstrated a reduced accumulation of Sb III due to an increase in drug efflux. Additionally, LABCG2 was able to transport thiols in the presence of Sb III . Biotinylation assays using parasites expressing LABCG2 fused with an N-terminal green fluorescent protein tag revealed that LABCG2 is partially localized in the plasma membrane; this supports data from previous studies which suggested that LABCG2 is localized in intracellular vesicles that fuse with the plasma membrane during exocytosis. In conclusion, Leishmania LABCG2 probably confers antimony resistance by sequestering metal-thiol conjugates within vesicles and through further exocytosis by means of the parasite's flagellar pocket.

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Section: Resultsmentioning

confidence: 99%

The LABCG2 Transporter from the Protozoan Parasite Leishmania Is Involved in Antimony Resistance

Perea

Manzano

Castanys

et al. 2016

Antimicrob Agents Chemother

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“…The development of RNA interference (RNAi) target sequencing (RIT-Seq) in kinetoplastid parasites, such as Trypanosoma brucei (6), has revealed numerous genes associated with drug action (7); however, RNAibased screening is not applicable to Leishmania, because most species lack functional RNAi machinery (8). In Leishmania spp., copy number variation and single-nucleotide polymorphism were detected in drug-resistant parasites using NGS (9, 10; reviewed in ref 11). Gain-of-function screening using genomic cosmid libraries is also a proven approach to studying drug resistance in Leishmania (12).…”

mentioning

confidence: 99%

Cos-Seq for high-throughput identification of drug target and resistance mechanisms in the protozoan parasite Leishmania

Gazanion

Fernández-Prada

Papadopoulou

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Innovative strategies are needed to accelerate the identification of antimicrobial drug targets and resistance mechanisms. Here we develop a sensitive method, which we term Cosmid Sequencing (or "Cos-Seq"), based on functional cloning coupled to next-generation sequencing. Cos-Seq identified >60 loci in the Leishmania genome that were enriched via drug selection with methotrexate and five major antileishmanials (antimony, miltefosine, paromomycin, amphotericin B, and pentamidine). Functional validation highlighted both known and previously unidentified drug targets and resistance genes, including novel roles for phosphatases in resistance to methotrexate and antimony, for ergosterol and phospholipid metabolism genes in resistance to miltefosine, and for hypothetical proteins in resistance to paromomycin, amphothericin B, and pentamidine. Several genes/loci were also found to confer resistance to two or more antileishmanials. This screening method will expedite the discovery of drug targets and resistance mechanisms and is easily adaptable to other microorganisms.

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“…This, together with the long treatment duration, leads to high abandonment rates. In addition, these drugs are effective only against some Leishmania species (De Menezes et al, 2015) and many cases of resistances have been described (Calvopina et al, 2006;Leprohon et al, 2015;Ubeda et al, 2014). The remarkable genomic plasticity of these parasites seems to play a central role in drug resistance.…”

Section: Drugs and Potential Targetsmentioning

confidence: 99%

“…Aneuploidy and single nucleotide polymorphisms are also widespread phenomena in Leishmania spp. (reviewed by Leprohon et al, 2015). All these events contribute to developing drug resistance.…”

Section: Drugs and Potential Targetsmentioning

confidence: 99%

Nuclear DNA replication and repair in parasites of the genusLeishmania: Exploiting differences to develop innovative therapeutic approaches

Uzcanga

Lara

Gutiérrez

et al. 2016

Critical Reviews in Microbiology

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Drug resistance analysis by next generation sequencing in Leishmania

Abstract: Graphical abstract

Cited by 71 publications

References 139 publications

The LABCG2 Transporter from the Protozoan Parasite Leishmania Is Involved in Antimony Resistance

The LABCG2 Transporter from the Protozoan Parasite Leishmania Is Involved in Antimony Resistance

Cos-Seq for high-throughput identification of drug target and resistance mechanisms in the protozoan parasite Leishmania

Nuclear DNA replication and repair in parasites of the genusLeishmania: Exploiting differences to develop innovative therapeutic approaches

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