Genomic and Metabolomic Polymorphism among Experimentally Selected Paromomycin-Resistant Leishmania donovani Strains

Shaw, Craig; Imamura, Hideo; Downing, Tim; Blackburn, Gavin; Westrop, Gareth D.; Cotton, James A.; Berriman, Matthew; Sanders, Mandy; Rijal, Suman; Coombs, Graham H.; Dujardin, Jean‐Claude; Carter, K. C.

doi:10.1128/aac.00904-19

Cited by 20 publications

(25 citation statements)

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“…The copyright holder for this preprint this version posted January 6, 2021. ; https://doi.org/10.1101/2021.01.05.425522 doi: bioRxiv preprint Whole genome sequencing is a powerful tool to identify molecular changes accompanying DR development in pathogens (7) (8,32). In this study, in-depth genomic analysis revealed that TCMDC-143345 resistant lines were characterized by several aneuploidy changes, CNVs (see discussion in supplementary text) and SNPs.…”

Section: Discussionmentioning

confidence: 87%

Genomic and phenotypic characterization of experimentally selected resistantLeishmania donovanireveals a role for dynamin-1 like protein in the mechanism of resistance to a novel anti-leishmanial compound

Hefnawy

Negreira

Jara

et al. 2021

Preprint

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The implementation of prospective drug resistance (DR) studies in the R&D pipelines is a common practice for many infectious diseases, but not for Neglected Tropical Diseases. Here, we explored and demonstrated the importance of this approach, using as paradigms Leishmania donovani, the etiological agent of Visceral Leishmaniasis (VL), and TCMDC-143345, a promising compound of the GSK ‘Leishbox’ to treat VL. We experimentally selected resistance to TCMDC-143345 in vitro and characterized resistant parasites at genomic and phenotypic levels. We found that it took more time to develop resistance to TCMDC-143345 than to other drugs in clinical use and that there was no cross resistance to these drugs, suggesting a new and unique mechanism. By whole genome sequencing, we found two mutations in the gene encoding the L. donovani dynamin-1-like protein (LdoDLP1) that were fixed at highest drug pressure. Through phylogenetic analysis, we identified LdoDLP1 as a family member of the dynamin-related proteins, a group of proteins that impacts the shapes of biological membranes by mediating fusion and fission events, with a putative role in mitochondrial fission. We found that L. donovani lines genetically engineered to harbor the two identified LdoDLP1 mutations were resistant to TCMDC-143345 and displayed altered mitochondrial properties. By homology modeling, we showed how the two LdoDLP1 mutations may influence protein structure and function. Taken together, our data reveal a clear involvement of LdoDLP1 in the adaptation/resistance of L. donovani to TCMDC-143345.ImportanceHumans and their pathogens are continuously locked in a molecular arms race during which the eventual emergence of pathogen drug resistance (DR) seems inevitable. For neglected tropical diseases (NTDs), DR is generally studied retrospectively, once it has already been established in clinical settings. We previously recommended to keep one step ahead in the host-pathogen arms race and implement prospective DR studies in the R&D pipeline, a common practice for many infectious diseases, but not for NTDs. Here, using Leishmania donovani, the etiological agent of Visceral Leishmaniasis (VL), and TCMDC-143345, a promising compound of the GSK ‘Leishbox’ to treat VL, as paradigms, we experimentally selected resistance to the compound and proceeded to genomic and phenotypic characterization of DR parasites. The results gathered in the present study suggest a new DR mechanism involving the L. donovani dynamin-1 like protein (LdoDLP1) and demonstrate the practical relevance of prospective DR studies.

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Section: Discussionmentioning

confidence: 87%

Genomic and phenotypic characterization of experimentally selected resistantLeishmania donovanireveals a role for dynamin-1 like protein in the mechanism of resistance to a novel anti-leishmanial compound

Hefnawy

Negreira

Jara

et al. 2021

Preprint

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“…Next, the antiparasitic activity of APC12, APC14 and APC16 against three Nepalese L. donovani clinical isolates with different inherent susceptibilities to antimony, reflective of strains present in endemic communities in the ISC, was tested. We found that the Sb-S, Sb-I and Sb-R [14] strains were also killed by the APC analogues and that efficacy was influenced by alkyl carbon chain length. APC16 and APC12 were the least and most toxic with death occurring below CMC for APC12-and APC14-treated Sb-S and Sb-I promastigotes, and above CMC for APC16-treated Sb-R promastigotes and amastigotes (Table 2 and Figure S2).…”

Section: The Effect Of Varied Alkyl Carbon Chain Lengths Of Apcs Agaimentioning

confidence: 89%

“…MHOM/NP/02/BPK087/0cl11 (Sb intermediate, Sb-I) and MHOM/NP/02/BPK275/0cl18 (Sb resistance, Sb-R [13]). In addition, Milt-resistant parasites raised against these three Sb sensitivity backgrounds [14] and the Sb-sensitive strain (MHOM/ET/67:LV82) was also used in studies. Luciferase expressing promastigotes for the LV82 and Nepalese strains were prepared using previously published methods and the integrative construct (a gift from Dr D.F.…”

Section: Animals and Parasitesmentioning

confidence: 99%

Structure and Antiparasitic Activity Relationship of Alkylphosphocholine Analogues against Leishmania donovani

2020

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Miltefosine (Milt) is the only oral treatment for visceral leishmaniasis (VL) but its use is associated with adverse effects, e.g., teratogenicity, vomiting, diarrhoea. Understanding how its chemical structure induces cytotoxicity, whilst not compromising its anti-parasitic efficacy, could identify more effective compounds. Therefore, we systemically modified the compound’s head, tail and linker tested the in vitro activity of three alkylphosphocholines (APC) series against Leishmania donovani strains with different sensitivities to antimony. The analogue, APC12, with an alkyl carbon chain of 12 atoms, was also tested for anti-leishmanial in vivo activity in a murine VL model. All APCs produced had anti-leishmanial activity in the micromolar range (IC50 and IC90, 0.46– > 82.21 µM and 4.14–739.89 µM; 0.01– > 8.02 µM and 0.09–72.18 µM, respectively, against promastigotes and intracellular amastigotes). The analogue, APC12 was the most active, was 4–10 fold more effective than the parent Milt molecule (APC16), irrespective of the strain’s sensitivity to antimony. Intravenous administration of 40 mg/kg APC12 to L. donovani infected BALB/c mice reduced liver and spleen parasite burdens by 60 ± 11% and 60 ± 19%, respectively, while oral administration reduced parasite load in the bone marrow by 54 ± 34%. These studies confirm that it is possible to alter the Milt structure and produce more active anti-leishmanial compounds.

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“…A Nepalese strain of L. donovani had a high natural resistance to PR and increased resistance to NO was noted. PR resistance was shown to be associated with lipidomic and metabolomic strain-specific changes [ 99 ]. A recent study using chemically induced mutagenesis identified a putative calcium dependent protein kinase (LinJ.33.1810) as a potential resistance marker [ 56 ].…”

Section: Benchmark Drugs For Leishmaniasismentioning

confidence: 99%

Can We Harness Immune Responses to Improve Drug Treatment in Leishmaniasis?

et al. 2020

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Leishmaniasis is a vector-borne parasitic disease that has been neglected in priority for control and eradication of malaria, tuberculosis, and HIV/AIDS. Collectively, over one seventh of the world’s population is at risk of being infected with 0.7–1.2 million new infections reported annually. Clinical manifestations range from self-healing cutaneous lesions to fatal visceral disease. The first anti-leishmanial drugs were introduced in the 1950′s and, despite several shortcomings, remain the mainstay for treatment. Regardless of this and the steady increase in infections over the years, particularly among populations of low economic status, research on leishmaniasis remains under funded. This review looks at the drugs currently in clinical use and how they interact with the host immune response. Employing chemoimmunotherapeutic approaches may be one viable alternative to improve the efficacy of novel/existing drugs and extend their lifespan in clinical use.

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Genomic and Metabolomic Polymorphism among Experimentally Selected Paromomycin-Resistant Leishmania donovani Strains

Cited by 20 publications

References 35 publications

Genomic and phenotypic characterization of experimentally selected resistantLeishmania donovanireveals a role for dynamin-1 like protein in the mechanism of resistance to a novel anti-leishmanial compound

Genomic and phenotypic characterization of experimentally selected resistantLeishmania donovanireveals a role for dynamin-1 like protein in the mechanism of resistance to a novel anti-leishmanial compound

Structure and Antiparasitic Activity Relationship of Alkylphosphocholine Analogues against Leishmania donovani

Can We Harness Immune Responses to Improve Drug Treatment in Leishmaniasis?

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