Diminazene resistance in <i>Trypanosoma congolense</i> is not caused by reduced transport capacity but associated with reduced mitochondrial membrane potential

Carruthers, Lauren V.; Munday, Jane C.; Ebiloma, Godwin U.; Steketee, Pieter C.; Jayaraman, Siddharth; Campagnaro, Gustavo Daniel; Ungogo, Marzuq A.; Lemgruber, Leandro; Donachie, Anne-Marie; Rowan, T.G.; Peter, Rose; Morrison, Liam J.; Barrett, Michael P.; Koning, Harry P. de

doi:10.1111/mmi.14733

Cited by 20 publications

(42 citation statements)

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“…It has been widely reported that in brucei group trypanosomes drug sensitivity and resistance is often linked to the presence or absence of particular transport proteins, such as TbAT1, TbAQP2, TbMFST and TbAAT6, which mediate the uptake of diamidines and melaminophenyl arsenicals (TbAT1 and TbAQP2), suramin (TbMFST) and eflornithine (TbAAT6), respectively [ 26 , 29 , 41 ]. However, the now well-established models of drug transport and resistance do not seem to apply to all African trypanosome species, as recently shown for diminazene uptake and resistance in T. congolense [ 42 ]. We here investigate whether the relatively low sensitivity of T. congolense for diamidines, melaminophenyl arsenicals and suramin is linked to the absence of orthologues of TbAT1, TbAQP2 and TbMFST.…”

Section: Resultsmentioning

confidence: 99%

Differences in Transporters Rather than Drug Targets Are the Principal Determinants of the Different Innate Sensitivities of Trypanosoma congolense and Trypanozoon Subgenus Trypanosomes to Diamidines and Melaminophenyl Arsenicals

Ungogo

Campagnaro

Al-Ghamdi

et al. 2022

IJMS

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The animal trypanosomiases are infections in a wide range of (domesticated) animals with any species of African trypanosome, such as Trypanosoma brucei, T. evansi, T. congolense, T. equiperdum and T. vivax. Symptoms differ between host and infective species and stage of infection and are treated with a small set of decades-old trypanocides. A complication is that not all trypanosome species are equally sensitive to all drugs and the reasons are at best partially understood. Here, we investigate whether drug transporters, mostly identified in T. b. brucei, determine the different drug sensitivities. We report that homologues of the aminopurine transporter TbAT1 and the aquaporin TbAQP2 are absent in T. congolense, while their introduction greatly sensitises this species to diamidine (pentamidine, diminazene) and melaminophenyl (melarsomine) drugs. Accumulation of these drugs in the transgenic lines was much more rapid. T. congolense is also inherently less sensitive to suramin than T. brucei, despite accumulating it faster. Expression of a proposed suramin transporter, located in T. brucei lysosomes, in T. congolense, did not alter its suramin sensitivity. We conclude that for several of the most important classes of trypanocides the presence of specific transporters, rather than drug targets, is the determining factor of drug efficacy.

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Section: Resultsmentioning

confidence: 99%

Differences in Transporters Rather than Drug Targets Are the Principal Determinants of the Different Innate Sensitivities of Trypanosoma congolense and Trypanozoon Subgenus Trypanosomes to Diamidines and Melaminophenyl Arsenicals

Ungogo

Campagnaro

Al-Ghamdi

et al. 2022

IJMS

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“…In much of sub-Saharan Africa, trypanosomiasis in (domesticated) animals is caused primarily by infection with T. congolense ; the condition is treated predominantly with diminazene aceturate but resistance to this drug is widespread [ 11 ]. Both the primary fractions of PNG propolis and the isolated compounds were tested on a standard drug-sensitive strain of T. congolense , IL3000, and the derived diminazene-resistant cell line 6C3 [ 29 ]. The crude extract displayed promising activity against T. congolense ( Table 6 and Table 7 ), including the resistant line, very close to the value obtained against T. b. brucei ( Table 3 ), but none of the individual fractions matched this.…”

Section: Resultsmentioning

confidence: 99%

“…There remain many triterpenoids to be isolated from the PNG propolis sample and there may be more active compounds present in the mixture given that its activity was higher than most of the isolated compounds. Importantly, 20-hydroxybetulin was significantly (>2-fold) active against drug-resistant strains of T. b. brucei and T. congolense , despite the drug resistance mechanisms being very different in the two species [ 29 , 30 ]. Overall, it would seem that 20-hydroxybetulin, which displayed low toxicity against mammalian cell lines, could be a promising lead compound to systematically explore the SAR against African drug-resistant trypanosomiasis.…”

Section: Discussionmentioning

confidence: 99%

“…The T. b. brucei strains were a standard drug-sensitive lab strain, Lister 427 (wild-type) [ 47 ] and the derived cell line B48 was developed from the wild-type by gene deletion of the drug transporter TbAT1 followed by in vitro adaptation to pentamidine [ 28 ], leading to the further loss of the gene encoding TbAQP2 [ 48 ], rendering it highly resistant to the diamidine and melaminophenyl arsenical classes of trypanocides. The T. congolense strains were the lab strain IL3000 and its diminazene-adapted clone 6C3 [ 29 ]. The C. fasciculata strain HS6 was a gift of Prof. Terry Smith (University of St Andrews, UK).…”

Section: Methodsmentioning

confidence: 99%

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The Antiprotozoal Activity of Papua New Guinea Propolis and Its Triterpenes

Alenezi

Ebiloma

et al. 2022

Molecules

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Profiling a propolis sample from Papua New Guinea (PNG) using high-resolution mass spectrometry indicated that it contained several triterpenoids. Further fractionation by column chromatography and medium-pressure liquid chromatography (MPLC) followed by nuclear magnetic resonance spectroscopy (NMR) identified 12 triterpenoids. Five of these were obtained pure and the others as mixtures of two or three compounds. The compounds identified were: mangiferonic acid, ambonic acid, isomangiferolic acid, ambolic acid, 27-hydroxyisomangiferolic acid, cycloartenol, cycloeucalenol, 24-methylenecycloartenol, 20-hydroxybetulin, betulin, betulinic acid and madecassic acid. The fractions from the propolis and the purified compounds were tested in vitro against Crithidia fasciculata, Trypanosoma congolense, drug-resistant Trypanosoma congolense, Trypanosoma b. brucei and multidrug-resistant Trypanosoma b. brucei (B48). They were also assayed for their toxicity against U947 cells. The compounds and fractions displayed moderate to high activity against parasitic protozoa but only low cytotoxicity against the mammalian cells. The most active isolated compound, 20-hydroxybetulin, was found to be trypanostatic when different concentrations were tested against T. b. brucei growth.

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“…Current methods of disease control centre around chemotherapy and prophylaxis (reviewed in [3]), but the very few available veterinary trypanocidal drugs have been used extensively for decades, resulting in resistance and inadequate protection [5][6][7][8]. In contrast to T. brucei [9], the resistance mechanisms of T. congolense are still poorly understood [10]. As such, there is a critical need for the development of new and improved chemotherapeutics to manage AAT [3,11], and furthering our knowledge of how T. congolense develops resistance to drugs can facilitate optimising the lifetime of both existing and new drugs.…”

Section: Introductionmentioning

confidence: 99%

Divergent metabolism between Trypanosoma congolense and Trypanosoma brucei results in differential sensitivity to metabolic inhibition

et al. 2021

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Animal African Trypanosomiasis (AAT) is a debilitating livestock disease prevalent across sub-Saharan Africa, a main cause of which is the protozoan parasite Trypanosoma congolense. In comparison to the well-studied T. brucei, there is a major paucity of knowledge regarding the biology of T. congolense. Here, we use a combination of omics technologies and novel genetic tools to characterise core metabolism in T. congolense mammalian-infective bloodstream-form parasites, and test whether metabolic differences compared to T. brucei impact upon sensitivity to metabolic inhibition. Like the bloodstream stage of T. brucei, glycolysis plays a major part in T. congolense energy metabolism. However, the rate of glucose uptake is significantly lower in bloodstream stage T. congolense, with cells remaining viable when cultured in concentrations as low as 2 mM. Instead of pyruvate, the primary glycolytic endpoints are succinate, malate and acetate. Transcriptomics analysis showed higher levels of transcripts associated with the mitochondrial pyruvate dehydrogenase complex, acetate generation, and the glycosomal succinate shunt in T. congolense, compared to T. brucei. Stable-isotope labelling of glucose enabled the comparison of carbon usage between T. brucei and T. congolense, highlighting differences in nucleotide and saturated fatty acid metabolism. To validate the metabolic similarities and differences, both species were treated with metabolic inhibitors, confirming that electron transport chain activity is not essential in T. congolense. However, the parasite exhibits increased sensitivity to inhibition of mitochondrial pyruvate import, compared to T. brucei. Strikingly, T. congolense exhibited significant resistance to inhibitors of fatty acid synthesis, including a 780-fold higher EC50 for the lipase and fatty acid synthase inhibitor Orlistat, compared to T. brucei. These data highlight that bloodstream form T. congolense diverges from T. brucei in key areas of metabolism, with several features that are intermediate between bloodstream- and insect-stage T. brucei. These results have implications for drug development, mechanisms of drug resistance and host-pathogen interactions.

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Diminazene resistance in Trypanosoma congolense is not caused by reduced transport capacity but associated with reduced mitochondrial membrane potential

Abstract: This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Cited by 20 publications

References 123 publications

Differences in Transporters Rather than Drug Targets Are the Principal Determinants of the Different Innate Sensitivities of Trypanosoma congolense and Trypanozoon Subgenus Trypanosomes to Diamidines and Melaminophenyl Arsenicals

Differences in Transporters Rather than Drug Targets Are the Principal Determinants of the Different Innate Sensitivities of Trypanosoma congolense and Trypanozoon Subgenus Trypanosomes to Diamidines and Melaminophenyl Arsenicals

The Antiprotozoal Activity of Papua New Guinea Propolis and Its Triterpenes

Divergent metabolism between Trypanosoma congolense and Trypanosoma brucei results in differential sensitivity to metabolic inhibition

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