Drug Target Identification Using a Trypanosome Overexpression Library

Begolo, Daniela; Erben, Esteban; Clayton, Christine

doi:10.1128/aac.03338-14

Cited by 33 publications

(39 citation statements)

References 27 publications

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“…Furthermore, half of the mutations were in the binding site of the drug’s molecular target. The high specificity of this approach is especially striking when compared with high-throughput approaches such as haploinsufficiency profiling, 35, 36 chemical pull-downs, 37 overexpression studies, 38 and other biochemical assays, which often implicate many genes rather than just the single target. 33 In addition, since drug selection theoretically queries all possible mutations, unlike haploinsufficiency profiling or biochemical pull-downs, it is able to provide a more specific answer while searching a much larger data set.…”

Section: Discussionmentioning

confidence: 99%

Rapid Chagas Disease Drug Target Discovery Using Directed Evolution in Drug-Sensitive Yeast

et al. 2016

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Recent advances in cell-based, high-throughput phenotypic screening have identified new chemical compounds that are active against eukaryotic pathogens. A challenge to their future development lies in identifying these compounds’ molecular targets and binding modes. In particular, subsequent structure-based chemical optimization and target-based screening require a detailed understanding of the binding event. Here we use directed evolution and whole-genome sequencing of a drug-sensitive S. cerevisiae strain to identify the yeast ortholog of TcCyp51, lanosterol-14-alpha-demethylase (TcCyp51), as the target of MMV001239, a benzamide compound with activity against Trypanosoma cruzi, the etiological agent of Chagas disease. We show that parasites treated with MMV0001239 phenocopy parasites treated with another TcCyp51 inhibitor, posaconazole, accumulating both lanosterol and eburicol. Direct drug-protein binding of MMV0001239 was confirmed through spectrophotometric binding assays and X-ray crystallography, revealing a binding site shared with other anti-trypanosomal compounds that target Cyp51. These studies provide a new probe chemotype for TcCyp51 inhibition.

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Section: Discussionmentioning

confidence: 99%

Rapid Chagas Disease Drug Target Discovery Using Directed Evolution in Drug-Sensitive Yeast

et al. 2016

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“…Drug resistance by gene overexpression is a useful approach for target identification and validation (33,34).Therefore, to test if CSN5 is the target of ZnDTC we examined the susceptibility of cells overexpressing CSN5. Overexpression of CSN5 resulted in increased To further evaluate whether ZnDTC acts in a COP9 dependent manner, we examined cullin1 deneddylation and UPS activity in drug-treated cells compared to controls.…”

Section: Zndtc Is Active Against E Histolytica In a Cop9-dependent Mmentioning

confidence: 99%

COP9 signalosome is an essential and druggable parasite target that regulates protein degradation

Ghosh

Farr

Singh

et al. 2020

Preprint

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Understanding how the protozoan protein degradation pathway is regulated could uncover new parasite biology for drug discovery. We found the COP9 signalosome (CSN) conserved in multiple pathogens such as Leishmania, Trypanosoma, Toxoplasma, and used the severe diarrhea-causing Entamoeba histolytica to study its function in medically significant protozoa. We show that CSN is an essential upstream regulator of parasite protein degradation. Genetic disruption of E. histolytica CSN by two distinct approaches inhibited cell proliferation and viability. Both CSN5 knockdown and dominant negative mutation trapped cullin in a neddylated state, disrupting UPS activity and protein degradation. In addition, zinc ditiocarb (ZnDTC), a main metabolite of the inexpensive FDA-approved alcohol-abuse drug disulfiram, was active against parasites acting in a COP9-dependent manner. ZnDTC, given as disulfiram-zinc, had oral efficacy in clearing parasites in vivo. Our findings provide insights into the regulation of parasite protein degradation, and supports the significant therapeutic potential of COP9 inhibition.Summary sentenceParasite-encoded COP9 signalosome is an essential upstream regulator of ubiquitin-proteasome mediated protein degradation, and shows significant potential as a therapeutic target.

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“…While this enzyme is ubiquitous in eukaryotes, including human isoforms, it has been validated as a target for cancer, as well as fungal, malarial, and viral infections . Furthermore, this enzyme has been found to be essential for kinetoplastid survival, including T. brucei in culture and in mouse models . While not completely resolved, over 60 proteins are believed to be substrates of TbNMT including ADP‐ribosylation factor‐1 (ARF‐1) and ADP‐ribosylation factor‐like (ARL‐1) protein, both of which are essential for bloodstream viability of T. brucei .…”

Section: Kinetoplastids (Trypanosomiasis and Leishmaniasis)mentioning

confidence: 99%

Unpacking the Pathogen Box—An Open Source Tool for Fighting Neglected Tropical Disease

Veale

2019

ChemMedChem

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The Pathogen Box is a 400‐strong collection of drug‐like compounds, selected for their potential against several of the world's most important neglected tropical diseases, including trypanosomiasis, leishmaniasis, cryptosporidiosis, toxoplasmosis, filariasis, schistosomiasis, dengue virus and trichuriasis, in addition to malaria and tuberculosis. This library represents an ensemble of numerous successful drug discovery programmes from around the globe, aimed at providing a powerful resource to stimulate open source drug discovery for diseases threatening the most vulnerable communities in the world. This review seeks to provide an in‐depth analysis of the literature pertaining to the compounds in the Pathogen Box, including structure–activity relationship highlights, mechanisms of action, related compounds with reported activity against different diseases, and, where appropriate, discussion on the known and putative targets of compounds, thereby providing context and increasing the accessibility of the Pathogen Box to the drug discovery community.

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Drug Target Identification Using a Trypanosome Overexpression Library

Cited by 33 publications

References 27 publications

Rapid Chagas Disease Drug Target Discovery Using Directed Evolution in Drug-Sensitive Yeast

Rapid Chagas Disease Drug Target Discovery Using Directed Evolution in Drug-Sensitive Yeast

COP9 signalosome is an essential and druggable parasite target that regulates protein degradation

Unpacking the Pathogen Box—An Open Source Tool for Fighting Neglected Tropical Disease

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